Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembl...Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp technique.Results: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2', 2'-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2001 Sep; 3: 185 - 191)展开更多
The patch clamp recording technique in vivois a blind patch clamp recording methods to record the current of the spinal or cereral neurons of anaes:hesia ( or awake) animals. This technique can be used to study the...The patch clamp recording technique in vivois a blind patch clamp recording methods to record the current of the spinal or cereral neurons of anaes:hesia ( or awake) animals. This technique can be used to study the synaptic function and plasticity in central nervous system in vivoin order to understand the physiological properties of the ion channels from an integrated point of view. The advantage of this technique have already presented itself in the study of the synaptic transmission and nervous network. Nowadays, in vivo patch whole-cell recording technique in combination with other techniques is becoming a common method in the research fields.展开更多
Interleukin-6 has been shown to be involved in nerve injury and nerve regeneration, but the effects of long-term administration of high concentrations of interleukin-6 on neurons in the central nervous system is poorl...Interleukin-6 has been shown to be involved in nerve injury and nerve regeneration, but the effects of long-term administration of high concentrations of interleukin-6 on neurons in the central nervous system is poorly understood. This study investigated the effects of 24 hour expo-sure of interleukin-6 on cortical neurons at various concentrations (0.1, 1, 5 and 10 ng/mL) and the effects of 10 ng/mL interleukin-6 exposure to cortical neurons for various durations (2, 4, 8, 24 and 48 hours) by studying voltage-gated Na+ channels using a patch-clamp technique. Volt-age-clamp recording results demonstrated that interleukin-6 suppressed Na+ currents through its receptor in a time- and dose-dependent manner, but did not alter voltage-dependent activation and inactivation. Current-clamp recording results were consistent with voltage-clamp recording results. Interleukin-6 reduced the action potential amplitude of cortical neurons, but did not change the action potential threshold. The regulation of voltage-gated Na+channels in rat corti-cal neurons by interleukin-6 is time- and dose-dependent.展开更多
Radiation therapy is a standard treatment for head and neck tumors.However,patients often exhibit cognitive impairments following radiation therapy.Previous studies have revealed that hippocampal dysfunction,specifica...Radiation therapy is a standard treatment for head and neck tumors.However,patients often exhibit cognitive impairments following radiation therapy.Previous studies have revealed that hippocampal dysfunction,specifically abnormal hippocampal neurogenesis or neuroinflammation,plays a key role in radiation-induced cognitive impairment.However,the long-term effects of radiation with respect to the electrophysiological adaptation of hippocampal neurons remain poorly characterized.We found that mice exhibited cognitive impairment 3 months after undergoing 10 minutes of cranial irradiation at a dose rate of 3 Gy/min.Furthermore,we observed a remarkable reduction in spike firing and excitatory synaptic input,as well as greatly enhanced inhibitory inputs,in hippocampal CA1 pyramidal neurons.Corresponding to the electrophysiological adaptation,we found reduced expression of synaptic plasticity marker VGLUT1 and increased expression of VGAT.Furthermore,in irradiated mice,long-term potentiation in the hippocampus was weakened and GluR1 expression was inhibited.These findings suggest that radiation can impair intrinsic excitability and synaptic plasticity in hippocampal CA1 pyramidal neurons.展开更多
Catilan extracted from Leiurus quinquestriatus is a specific ion channel blocker.It can specifically bind chloride channels of glioma cells and kill these tumor cells.The questions remain as to whether antigliomatin,t...Catilan extracted from Leiurus quinquestriatus is a specific ion channel blocker.It can specifically bind chloride channels of glioma cells and kill these tumor cells.The questions remain as to whether antigliomatin,the extract from scorpion venom of Buthus martensii Karsch in China,can inhibit glioma growth,and whether this inhibition is correlated with ion channels of tumor cells.The present study treated rat C6 glioma cells with 0.8,1.2,and 1.6 μg/mL antigliomatin for 20 hours.Whole-cell patch clamp technique showed that antigliomatin delayed rectifier potassium channels of C6 glioma cells.Antigliomatin inhibited tumor growth,which could potentially involve potassium channels of tumor cells.展开更多
MicroRNAs can regulate the function of ion channels in many organs.Based on our previous study we propose that miR-142a-39,which is highly expressed in denervated skeletal muscle,might affect cell excitability through...MicroRNAs can regulate the function of ion channels in many organs.Based on our previous study we propose that miR-142a-39,which is highly expressed in denervated skeletal muscle,might affect cell excitability through similar mechanisms.In this study,we overexpressed or knocked down miR-142a-3p in C2C12 cells using a lentivirus method.After 7 days of differentiation culture,whole-cell currents were recorded.The results showed that overexpression of miR-142a-3p reduced the cell membrane capacitance,increased potassium current density and decreased calcium current density.Knockdown of miR-142a-3p reduced sodium ion channel current density.The results showed that change in miR-142a-3p expression affected the ion channel currents in C2C12 cells,suggesting its possible roles in muscle cell electrophysiology.This study was approved by the Animal Ethics Committee of Peking University in July 2020(approval No.LA2017128).展开更多
BACKGROUND: Gamma-knife is characterized by low risk and low death rate, without trauma, bleeding or infection. MRI has replaced CT as a method of location, and provides a good choice for treating pituitary tumors.OB...BACKGROUND: Gamma-knife is characterized by low risk and low death rate, without trauma, bleeding or infection. MRI has replaced CT as a method of location, and provides a good choice for treating pituitary tumors.OBJECTIVE: To analyze the effectiveness and complications of 501 cases with pituitary adenoma by using gamma-knife retrospectively.DESIGN: Case-analysis.SETTING: Gamma-knife Center, the 363 Hospital of Chinese PLA.PARTICIPANTS: A total of 501 cases were selected from Gamma-knife Center, the 363 Hospital of Chinese PLA from January 1997 to December 2002. All patients were certainly diagnosed with CT and MRI scanning. There were 186 males and 315 females. Their ages ranged from 15 to 84 years with the mean age of 39 years. All patients provided confirmed consent.METHODS: All 501 cases with pituitary adenomas were treated by gamma-knife or combined with operations: The average iso-dose curve was 50% (30% - 65%), peri-dose was 14 Gy and the average target number was 5.17. ① At 35 months after treatment, patients received follow-up including clinical symptoms,imaging symptoms and endocrine symptoms. ② Patients who received second gamma-knife treatment were analyzed and their complications were observed after operation.MAIN OUTCOME MEASURES: ① Follow-up results of clinical symptoms, image and endocrine indexes; ② second gamma-knife treatment; ③ postoperative complications.RESULTS: Among 501 accepted patients, 275 cases were involved in the follow up of clinical symptoms,154 in image symptoms and 98 in endocrine symptoms. ① Follow-up results of clinical symptoms, image and endocrine indexes: Follow up of clinical symptoms demonstrated that clinical symptoms of 169 cases were relieved, of 68 disappeared, and of 38 deteriorated. Follow-up of image symptoms indicated that pituitary tumor of 25 cases disappeared, of 84 shortened, of 42 not changed, and of 3 enlarged. Follow-up of endocrine symptoms suggested that endocrine of 50 cases was abnormal, of 29 recovery to normal value, and of 19 not changed obviously. ② Second gamma-knife treatment: At 6- 24 months after gamma-knife treatment, 15 patients with pituitary tumor received second gamma-knife treatment; especially, one patient received for the three times. The conditions in details were recorded as follows: The peri-dose for the first treatment ranged from 8 to 12 Gy; in addition, that for the second one ranged from 6 to 12 Gy. ③ Postoperative complications: The complications were as follow: hypopituitarism occurred in 2 patients (0.7%); tumor apoplexy in 2 patients (0.7%);weakened eyesight in 5 patients (1.8%); 3 of which recovered with dehydration and hormone treatment; 2 patients received tumor resection and optical nerve decompression operation with dehydration and hormone treatment ineffectively.CONCLUSION: Gamma-knife is effective and safe for pituitary adenoma.展开更多
BACKGROUND: Induced differentiation strategies and cytochemical properties of human embryonic stem ceils (hESCs) have been investigated. However, the electrophysiological functions of tyrosine hydroxylase (TH)-po...BACKGROUND: Induced differentiation strategies and cytochemical properties of human embryonic stem ceils (hESCs) have been investigated. However, the electrophysiological functions of tyrosine hydroxylase (TH)-positive cells dedved from hESCs remain unclear. OBJECTIVE: To investigate the differentiation efficiency of TH-positive cells from hESCs in vitro using modified four-step culture methods, including embryoid body formation, and to examine the functional characteristics of the differentiated TH-positive cells using electrophysiological techniques. DESIGN, TIME AND SETTING: Neuroelectrophysiology was performed at the Reproductive Medicine Center and Stem Cell Research Center, Peking University Third Hospital, and the Neuroscience Research Institute and Department of Neurobiology, Peking University, from September 2004 to August 2008. MATERIALS: The hESC line, PKU-1.1, a monoclonal cell line derived from a pre-implantation human blastocyst in the Reproductive Medical Center of Peking University Third Hospital. The patch clamp recording system was provided by the Neuroscience Research Institute and Department of Neurobiology, Peking University. METHODS: The hESC line was induced to differentiate into TH-positive cells in vitro using a modified four-step culture method, including the formation of embryoid body, as well as the presence of sonic hedgehog and fibroblast growth factor 8. The cell karyotype was assessed by G-banding karyotype analysis techniques and specific markers were detected immunocytochemically. Whole-cell configuration was obtained after obtaining a tight seal of over 1 GΩ. Ionic currents were detected by holding the cells at -70 mV and stepping to test voltages between -80 and 40 mV in 10-mV increments in voltage-clamp configuration. MAIN OUTCOME MEASURES: We measured the cell karyotype, specific cell markers, and the electrophysiological properties of the voltage-gated ion channels on the cell membrane of TH-positive dopaminergic cells differentiated from our hESCs line in vitro. RESULTS: The differentiated cells had a consistent appearance, and the majority of cells (〉 90%) expressed TH and β-tubulion, as well as the neural progenitor marker, nestino Cell karyotype analysis demonstrated that all of the hESCs had a stable and normal karyotype (46, XX) after differentiation. In addition, patch clamp recording showed that the 10 recorded TH-positive cells exhibited a fast inward current when the test voltage depolarized to -30 mV, and a delayed outward current when the test voltage depolarized to -10 mV. The peak of inward current was obtained at voltage between 10 mV and 0 mV, while the peak of outward current was obtained at 40 mV. The average peak of inward current density was ( -50.05 ± 15.50) pA/pF, and the average peak of outward current density was (41.98 ± 13.55) pA/pE CONCLUSION: More than 90% of the differentiated hESC-derived cells induced by the modified four-step culture method exhibit dopaminergic neuronal properties, including general electrophysiological functional properties, such as functional potassium and sodium channels.展开更多
The γ-aminobutyric acid neurotransmitter in the spinal cord dorsal horn plays an important role in pain modulation through primary afferent-mediated presynaptic inhibition. The weakening of γ-aminobutyric acid-media...The γ-aminobutyric acid neurotransmitter in the spinal cord dorsal horn plays an important role in pain modulation through primary afferent-mediated presynaptic inhibition. The weakening of γ-aminobutyric acid-mediated presynaptic inhibition may be an important cause of neuropathic pain. γ-aminobutyric acid-mediated presynaptic inhibition is related to the current strength of γ-aminobutyric acid A receptor activation. In view of this, the whole-cell patch-clamp technique was used here to record the change in muscimol activated current of dorsal root ganglion neurons in a chronic constriction injury model. Results found that damage in rat dorsal root ganglion neurons following application of muscimol caused concentration-dependent activation of current, and compared with the sham group, its current strength and γ-aminobutyric acid A receptor protein expression decreased. Immunofluorescence revealed that γ-aminobutyric acid type A receptor α2 subunit protein expression decreased and was most obvious at 12 and 15 days after modeling. Our experimental findings confirmed that the y-aminobutyric acid type A receptor α2 subunit in the chronic constriction injury model rat dorsal root ganglion was downregulated, which may be one of the reasons for the reduction of injury in dorsal root ganglion neurons following muscimol-activated currents.展开更多
Using whole-cell patch-clamp technique, the present study examined the effects of captopril on membrane currents of ventricular myocytes. The results were as follows: when the concentration of captopril was 6×10-...Using whole-cell patch-clamp technique, the present study examined the effects of captopril on membrane currents of ventricular myocytes. The results were as follows: when the concentration of captopril was 6×10-4 mol, the fast inward sodium current and slow inward calcium current were all decreased significantly (P<0.05). Besides, when the concentration exceeds 10-3 mol,the delayed rectifier potassium current was increased (P<0. 05) as well. But if the experimental concentration was under 6×10-4 mol, these three kinds of currents was not affected. These findings suggested that captopril has no direct antiarrhythmic effect at the average therapeutic serum level.展开更多
The roles of voltage-dependent K^+ channels during activation and damage in alveolar macrophages (AMs) exposed to different silica particles were examined. Rat AMs were collected by means of bronchoalveolar lavage,...The roles of voltage-dependent K^+ channels during activation and damage in alveolar macrophages (AMs) exposed to different silica particles were examined. Rat AMs were collected by means of bronchoalveolar lavage, and were adjusted to 5× 10^5/mL. After AMs were exposed to different concentrations (0, 25, 50, 100, 200 μg/mL) of quartz particles and 100 μg/mL amorphous silica particles for 24 h, the voltage-depended K^+ current in AMs was measured by using patch clamp technique. Meanwhile the leakage of lactate dehydrogenase (LDH) and the viability of AMs were detected respectively. Patch clamp studies demonstrated that AMs possessed outward delayed and inward rectifying K^+ current. Exposure to quartz particles increased the outward delayed K^+ current but it had no effect on inward rectifier K^+ current in AMs. Neither of the two K^+ channels in AMs was affected by amorphous silica particles. Cytotoxicity test showed that both silica particles could damage AM membrane and result in significant leakage of LDH (P〈0.05). MTT studies, however, showed that only quartz particles reduced viability of AMs (P〈0.05). It is concluded that quartz parti- cles can activate the outward delayed K^+ channel in AMs, which may act as an activating signal in AMs to initiate an inflammatory response during damage and necrosis in AMs induced by exposure to quartz particle. K^+ channels do not contribute to the membrane damage of AMs.展开更多
Objective: Heart failure(HF), a worldwide health condition, is the result of many cardiovascular diseases.The traditional Chinese medicine(TCM) Xiaoyu Jiangzhi capsule(XYC) has long been in use in China to treat hyper...Objective: Heart failure(HF), a worldwide health condition, is the result of many cardiovascular diseases.The traditional Chinese medicine(TCM) Xiaoyu Jiangzhi capsule(XYC) has long been in use in China to treat hyperlipidemia and inhibit platelet aggregation. This study explores the effects of XYC on heart failure(HF) and its detailed mechanisms.Methods: Isoproterenol(ISO, 30 mg/kg) was injected intraperitoneally for 7 days to copy a HF model of 10-12 weeks old, 20-30 g male mice. We then compared the CON(control) group, ISO(HF model)group, MET(metoprolol) group, and XYC group. Cardiac systolic function and left wall thickness were evaluated by echocardiograph. Using western blot analysis, we detected the proteins of calmodulin dependent protein kinase Ⅱ(Ca MKII) and sarco/endoplasmic reticulum Ca^(2+)-ATPase(Serca). Furthermore, ts A201 cells were cultured and the human CaV1.2 calcium channel current(hCaV1.2) were detected by patch clamp experiments.Results: XYC reduced HF, inhibiting the protein expression of Ca MKII, but Serca did not change significantly. Moreover, XYC inhibited the peak amplitude of the hCaV1.2 current, depolarizing shifted the activation curve 27.6 mV, and shifted the inactivation curve toward a positive potential 17.6 mV. The fraction recovered from inaction was reduced in XYC group compared with that in CON group.Conclusion: XYC could inhibit ISO-induced HF by reducing the Ca^(2+)/Ca MKII signaling pathway in mice.展开更多
Chlorogenic acid(5-caffeoylquinic acid, CGA) is a phenolic compound that is found ubiquitously in plants, fruits and vegetables and is formed via the esterification of caffeic acid and quinic acid. In addition to it...Chlorogenic acid(5-caffeoylquinic acid, CGA) is a phenolic compound that is found ubiquitously in plants, fruits and vegetables and is formed via the esterification of caffeic acid and quinic acid. In addition to its notable biological functions against cardiovascular diseases, type-2 diabetes and inflammatory conditions, CGA was recently hypothesized to be an alternative for the treatment of neurological diseases such as Alzheimer's disease and neuropathic pain disorders. However, its mechanism of action is unclear.Voltage-gated potassium channel(Kv) is a crucial factor in the electro-physiological processes of sensory neurons. Kv has also been identified as a potential therapeutic target for inflammation and neuropathic pain disorders. In this study, we analysed the effects of CGA on the two main subtypes of Kv in trigeminal ganglion neurons, namely, the IK,Aand IK,Vchannels. Trigeminal ganglion(TRG)neurons were acutely disassociated from the rat TRG, and two different doses of CGA(0.2 and 1 mmol·L21) were applied to the cells.Whole-cell patch-clamp recordings were performed to observe alterations in the activation and inactivation properties of the IK,Aand IK,Vchannels. The results demonstrated that 0.2 mmol·L21CGA decreased the peak current density of IK,A. Both 0.2 mmol·L21and1 mmol·L21CGA also caused a significant reduction in the activation and inactivation thresholds of IK,Aand IK,V. CGA exhibited a strong effect on the activation and inactivation velocities of IK,Aand IK,V. These findings provide novel evidence explaining the biological effects of CGA, especially regarding its neurological effects.展开更多
The study examined the inhibitory effect of Atractylodes macrocephala (AM) on the uterine contraction during premature delivery and explored its electrophysiological mechanism by studying the effects of AM on the C...The study examined the inhibitory effect of Atractylodes macrocephala (AM) on the uterine contraction during premature delivery and explored its electrophysiological mechanism by studying the effects of AM on the Ca^2+-activated K^+ currents of pregnant human myometrial smooth muscle cells with or without the treatment with intedeukin-6. Single cells were acutely isolated from pregnant human myometrial smooth muscles. Whole-cell Ca^2+-activated K^+ currents were recorded by using an Axopatchl-D amplifier. The cells were divided into three groups: group A in which AM was added into perfusate, group B, in which interleukin-6 was added into perfusate) and group C in which AM was added into perfusate after addition of interleukin-6. IL-6 10 ng/mL inhibited BKca by 36.9%±13.7% as compared with control (P〈0.01). AM at 2 mg/mL raised BKca by 36.7%±22.6% or 45.2%±13.7% with or without the treatment of IL-6, respectively (P〈0.01). It is concluded that AM was able to enhance the BKca of pregnant human myometrial smooth muscle cells treated or untreated with interleukin-6 and its effect on the BKca IL-treated cells was stronger that its effect on BKca of untreated cells. Our results suggested that AM can help to maintain the membrane potentials and the resting status of pregnant human myometrial smooth muscle cells.展开更多
Summary: To investigate the exact mechanism of epileptogenesis induced by coriaria lactone (CL), the effect of CL on NMDA receptor mediated current (IAsp) in rat hippocampal CA1 neu- rons was investigated by using ny...Summary: To investigate the exact mechanism of epileptogenesis induced by coriaria lactone (CL), the effect of CL on NMDA receptor mediated current (IAsp) in rat hippocampal CA1 neu- rons was investigated by using nystatin perforated whole-cell patch clamp. 10-6-10-4 mol/L Asp acted on NMDA receptors and elicited an inward current (IAsp) at a holding potential (VH) of -40 mV in presence of 10-6 mol/L glycine and absence of Mg2+ extracellularly. CL enhanced NMDA receptor mediated current induced by Asp, but had no effect on threshold concentration, EC50, Hill coefficient as well as maximal-effect concentration and reversal potential of IAsp. The effect had no relationship with holding potential. These results showed that CL could enhance NMDA receptor mediated current to increase [Ca2+]i of neurons by acting on Gly site, thereby inducing epilepsy.展开更多
Objective:To study the effects of sodium ferulate on the ultrarapid delayed rectifier K^+ current(IKur) in human atrial myocytes. Methods:Human atrial myocytes were isolated by enzyme dispersion method. IKur, in ...Objective:To study the effects of sodium ferulate on the ultrarapid delayed rectifier K^+ current(IKur) in human atrial myocytes. Methods:Human atrial myocytes were isolated by enzyme dispersion method. IKur, in human atrial myocytes were recorded by using the whole cell patch clamp. The changes of IKur were compared in the absence and the presence of sodium ferulate. Results:There was no effect of 0.4 g/L sodium ferulate on I-V relation of IKur. However, 0.4 g/L sodium ferulate inhibited IKur to some degrees at each test pulse. The current densities of IKur at +60 mV decreased from 4.997 ± 0.35 PA/PF to 3.331 ± 0.26 PA/PF(n = 6, P 〈 0.05). The inhibitory effect was concentration-dependent. IC50 was(0.41 ±0.03)g/L and the Hill coefficient was 0.95 ± 0.05. Conclusion:Sodium ferulate as a potassium channel blocker can inhibit IKur in human atrial myocytes effectively.展开更多
BACKGROUND: Some experiments have demonstrated that injecting orphanin FQ (OFQ) into lateral ventricle, which can obviously decrease the pain threshold. It is indicated that OFQ is an anti-opiate substance. However...BACKGROUND: Some experiments have demonstrated that injecting orphanin FQ (OFQ) into lateral ventricle, which can obviously decrease the pain threshold. It is indicated that OFQ is an anti-opiate substance. However, whether OFQ has effects on sensory neuron ion channel in cerebral cortex needs to be further studied. OBJECTIVE: To investigate the effects of OFQ, morphine or their combination on sodium channel current of somatosensory neurons in rat cerebral cortex. DESIGN: Repeated measurement trial. SETTING: Department of Physiology, Harbin Medical University. MATERIALS: Fifty healthy Wistar rats, aged 12 - 16 days, of either gender, were provided by the Experimental Animal Center, Second Hospital Affiliated to Harbin Medical University. OFQ was purchased from Sigma-Aldrich Company, and morphine was provided by the Shenyang First Pharmaceutical Factory. PC2C patch clamp amplifier and LabmasterTLlwere purchased from Yibo Life Science Instrument Co.,Ltd. of Huazhong University of Science and Techgnology. METHODS: This experiment was carried out in the Department of Physiology (provincial laboratory), Harbin Medical University between January 2005 and May 2006. Cortical neurons were acutely isolated from rats, and prepared into cell suspension following culture. ①Sodium channel current of somatosensory neurons in rat cerebral cortex was recorded before and after administration by whole-cell Patch clamp technique after 50 nmol/L OFQ being added to extracellular fluid. ②The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex was recorded before and after administration by the same method after 20 μmol/L morphine being added to extracellular fluid, and then the change of sodium channel current was recorded after 50 nmol/L OFQ being added. MAIN OUTCOME MEASURES: The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex following the administration of OFQ, morphine separately or their combination.. RESULTS: ①The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex was significantly lower after administration of 50 nmol/L OFQ than before at the clampe of the voltage of -30 mV (P 〈 0.05).②The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex was significantly lower after administration of 20 μmol/L morphine than before at the clampe of the voltage of-30 mV (P 〈 0.05). The sodium channel current recovered to - (2 345.24±174.18) pA after 50 nmol/L OFQ was administrated. There were significant differences in the amplitude of Na^+ channel current between two interventions (P 〈 0.05). CONCLUSION: Morphine and OFQ can respectively reduce the amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex, and OFQ can reverse the effect of morphine partly. It is indicated that OFQ can produce antiopioid activity in the central nervous system by influencing sodium channel current.展开更多
Cannabinoid and nicotinic acetylcholine receptors are strongly associated with algesia. Previous studies in our laboratory have reported inhibitory effects of synthetic cannabinoid WIN55, 212-2 on nicotine-activated c...Cannabinoid and nicotinic acetylcholine receptors are strongly associated with algesia. Previous studies in our laboratory have reported inhibitory effects of synthetic cannabinoid WIN55, 212-2 on nicotine-activated currents (Inic), but the underlying mechanisms remain poorly understood. The present study used whole-cell patch clamp techniques to investigate the modulatory effects of synthetic cannabinoid WIN55, 212-2 on Inic in cultured rat trigeminal ganglion neurons. The results revealed several major findings: WIN55, 212-2 inhibited Inic in rat trigeminal ganglion neurons. In addition, when WIN55, 212-2 (3 μmol/L) was applied simultaneously with nicotine (100 μmol/L), the inhibition of WIN55, 212-2 on Inic was reversible, concentration-dependent and voltage-independent This effect was not mediated by CB1, CB2 or VR1 receptors; neither the selective CB1 receptor antagonist AM281, CB2 receptor antagonist AM630 nor VR1 receptor antagonist capsazepine reduced the inhibitory effect of WIN55, 212-2. Further, the inhibition of nicotinic responses by WIN55, 212-2 was not sensitive to the membrane permeable cyclic adenosine monophosphate (cAMP) analog 8-Br-cAMP. The G-protein inhibitor GDP-I3-S (1 mmol/L) did not block the inhibitory effects of WIN55, 212-2 on/n^c, excluding the involvement of G-protein mediation. The results suggested that WIN55, 212-2 inhibits/n^o directly via the neuronal nicotinic acetylcholine receptor, and that this inhibition is non-competitive. WIN55, 212-2 did not act as an open channel blocker of the neuronal nicotinic acetylcholine receptor, and did not affect the desensitization of Into. The results suggest that nicotine receptors may be physically plugged from outside the membrane by drugs containing WIN55, 212-2.展开更多
OBJECTIVE Toad venom(Venenum Bufonis)isalways used for analgesia in China from ancient to modern times,but the effective component of it remains unclear.METHODS In the present study,we investigated the anti-nociceptiv...OBJECTIVE Toad venom(Venenum Bufonis)isalways used for analgesia in China from ancient to modern times,but the effective component of it remains unclear.METHODS In the present study,we investigated the anti-nociceptive effect and the underlying mechanism ofbufalin,an active ingredient fromtoad venom by animal behavior,patch clamp and calcium imaging.RESULTS Bufalin could significantly relieve formalin-induced spontaneous flinching and licking response as well as carrageenan-induced mechanical and thermal hyperalgesia.Using the whole-cel patch-clamp recording,bufalincaused remarkable suppressive effect on the peak currents of Na+channels in dorsal root ganglion neuroblastoma ND7-23 cel line in a U-shaped dependent manner.In addition,bufalinprompted the voltage-dependent activationand caused a negative shift of the fast-state inactivation of Na+channels.However,bufalin produced insignificant effect not onlyon voltage-dependent Kv4.2,Kv4.3 and BK channels,but also on the capsaicin induced Ca2+influx.CONCLUSION The present results indicate bufalin is capable of producing remarkable anti-nociceptive effects whichis probably ascribed to its specific modulation of voltage-gated Na+channels.展开更多
The current difference between male and female rabbit ventricular myocytes was investigated for elucidating the mechanism of longer QT interval and higher incidence of drug-associated torsade de pointes in female rabb...The current difference between male and female rabbit ventricular myocytes was investigated for elucidating the mechanism of longer QT interval and higher incidence of drug-associated torsade de pointes in female rabbits than in male rabbits. Whole cell patch clamp technique was used to record APD, I_to, I_K,tail, I_K1 and I_Ca,L of myocytes from left ventricular apex. There was no difference in the membrane capacitance between male and female rabbit myocytes. APD_90 was longer in female rabbits (560.4±26.5 ms, n=15) than in male ones (489.0±20.7 ms, n=14), P<0.05. In female rabbit myocytes, I_K,tail, I_to, I_K1 and I_Ca,L were 0.71±0.05 pA/pF (n=17), 8.28±1.03 pA/pF (n=18), 24.5±3.6 pA/pF (n=12) and 9.0±2.3 pA/pF (n=15) respectively. In male rabbit myocytes, they were 0.84±0.07 pA/pF (n=18), 8.60±1.20 pA/pF (n=18), 25.9±4.5 pA/pF (n=14) and 9.3±2.6 pA/pF (n=16) respectively. I_K,tail in female rabbits was significantly lower than that of male rabbits (P<0.05), but there was no difference in I_to, I_K1 and I_Ca,L between male rabbits and female rabbits (P>0.05). The lower I_K,tail of female rabbit myocytes may contribute to the longer repolarization and the higher incidence of drug-associated torsade de pointes.展开更多
文摘Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp technique.Results: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2', 2'-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2001 Sep; 3: 185 - 191)
文摘The patch clamp recording technique in vivois a blind patch clamp recording methods to record the current of the spinal or cereral neurons of anaes:hesia ( or awake) animals. This technique can be used to study the synaptic function and plasticity in central nervous system in vivoin order to understand the physiological properties of the ion channels from an integrated point of view. The advantage of this technique have already presented itself in the study of the synaptic transmission and nervous network. Nowadays, in vivo patch whole-cell recording technique in combination with other techniques is becoming a common method in the research fields.
基金supported by a grant from the National Natural Science Foundation of China,No.30972766,31170852,81001322,81172795,81173048the Specialized Research Fund for the Doctoral Program of Colleges and Universities,No.20094402110004
文摘Interleukin-6 has been shown to be involved in nerve injury and nerve regeneration, but the effects of long-term administration of high concentrations of interleukin-6 on neurons in the central nervous system is poorly understood. This study investigated the effects of 24 hour expo-sure of interleukin-6 on cortical neurons at various concentrations (0.1, 1, 5 and 10 ng/mL) and the effects of 10 ng/mL interleukin-6 exposure to cortical neurons for various durations (2, 4, 8, 24 and 48 hours) by studying voltage-gated Na+ channels using a patch-clamp technique. Volt-age-clamp recording results demonstrated that interleukin-6 suppressed Na+ currents through its receptor in a time- and dose-dependent manner, but did not alter voltage-dependent activation and inactivation. Current-clamp recording results were consistent with voltage-clamp recording results. Interleukin-6 reduced the action potential amplitude of cortical neurons, but did not change the action potential threshold. The regulation of voltage-gated Na+channels in rat corti-cal neurons by interleukin-6 is time- and dose-dependent.
基金supported by the National Natural Science Foundation of China,Nos.81925031(to YT),81820108026(to YT),81972967(to WJL),81872549(to YL)the Youth Program of National Natural Science Foundation of China,No.81801229(to YTX)+3 种基金a grant from Guangdong Science and Technology Department of China,Nos.2020B1212060018(to WJL),2020B1212030004(to WJL)the Natural Science Foundation of Guangdong Province,No.2019A1515011754(to WJL)the Science and Technology Program of Guangzhou of China,No.202007030001(to YT)the Science and Technology Planning Project of Guangzhou of China,No.201704030033(to YL).
文摘Radiation therapy is a standard treatment for head and neck tumors.However,patients often exhibit cognitive impairments following radiation therapy.Previous studies have revealed that hippocampal dysfunction,specifically abnormal hippocampal neurogenesis or neuroinflammation,plays a key role in radiation-induced cognitive impairment.However,the long-term effects of radiation with respect to the electrophysiological adaptation of hippocampal neurons remain poorly characterized.We found that mice exhibited cognitive impairment 3 months after undergoing 10 minutes of cranial irradiation at a dose rate of 3 Gy/min.Furthermore,we observed a remarkable reduction in spike firing and excitatory synaptic input,as well as greatly enhanced inhibitory inputs,in hippocampal CA1 pyramidal neurons.Corresponding to the electrophysiological adaptation,we found reduced expression of synaptic plasticity marker VGLUT1 and increased expression of VGAT.Furthermore,in irradiated mice,long-term potentiation in the hippocampus was weakened and GluR1 expression was inhibited.These findings suggest that radiation can impair intrinsic excitability and synaptic plasticity in hippocampal CA1 pyramidal neurons.
基金the Science and Technology Development Program of Jilin Province, No. 20050407-6
文摘Catilan extracted from Leiurus quinquestriatus is a specific ion channel blocker.It can specifically bind chloride channels of glioma cells and kill these tumor cells.The questions remain as to whether antigliomatin,the extract from scorpion venom of Buthus martensii Karsch in China,can inhibit glioma growth,and whether this inhibition is correlated with ion channels of tumor cells.The present study treated rat C6 glioma cells with 0.8,1.2,and 1.6 μg/mL antigliomatin for 20 hours.Whole-cell patch clamp technique showed that antigliomatin delayed rectifier potassium channels of C6 glioma cells.Antigliomatin inhibited tumor growth,which could potentially involve potassium channels of tumor cells.
基金supported by the National Natural Science Foundation of China,Nos.82072162,81971177Beijing Municipal Natural Science Foundation of China,No.7192215(all to XFY)。
文摘MicroRNAs can regulate the function of ion channels in many organs.Based on our previous study we propose that miR-142a-39,which is highly expressed in denervated skeletal muscle,might affect cell excitability through similar mechanisms.In this study,we overexpressed or knocked down miR-142a-3p in C2C12 cells using a lentivirus method.After 7 days of differentiation culture,whole-cell currents were recorded.The results showed that overexpression of miR-142a-3p reduced the cell membrane capacitance,increased potassium current density and decreased calcium current density.Knockdown of miR-142a-3p reduced sodium ion channel current density.The results showed that change in miR-142a-3p expression affected the ion channel currents in C2C12 cells,suggesting its possible roles in muscle cell electrophysiology.This study was approved by the Animal Ethics Committee of Peking University in July 2020(approval No.LA2017128).
文摘BACKGROUND: Gamma-knife is characterized by low risk and low death rate, without trauma, bleeding or infection. MRI has replaced CT as a method of location, and provides a good choice for treating pituitary tumors.OBJECTIVE: To analyze the effectiveness and complications of 501 cases with pituitary adenoma by using gamma-knife retrospectively.DESIGN: Case-analysis.SETTING: Gamma-knife Center, the 363 Hospital of Chinese PLA.PARTICIPANTS: A total of 501 cases were selected from Gamma-knife Center, the 363 Hospital of Chinese PLA from January 1997 to December 2002. All patients were certainly diagnosed with CT and MRI scanning. There were 186 males and 315 females. Their ages ranged from 15 to 84 years with the mean age of 39 years. All patients provided confirmed consent.METHODS: All 501 cases with pituitary adenomas were treated by gamma-knife or combined with operations: The average iso-dose curve was 50% (30% - 65%), peri-dose was 14 Gy and the average target number was 5.17. ① At 35 months after treatment, patients received follow-up including clinical symptoms,imaging symptoms and endocrine symptoms. ② Patients who received second gamma-knife treatment were analyzed and their complications were observed after operation.MAIN OUTCOME MEASURES: ① Follow-up results of clinical symptoms, image and endocrine indexes; ② second gamma-knife treatment; ③ postoperative complications.RESULTS: Among 501 accepted patients, 275 cases were involved in the follow up of clinical symptoms,154 in image symptoms and 98 in endocrine symptoms. ① Follow-up results of clinical symptoms, image and endocrine indexes: Follow up of clinical symptoms demonstrated that clinical symptoms of 169 cases were relieved, of 68 disappeared, and of 38 deteriorated. Follow-up of image symptoms indicated that pituitary tumor of 25 cases disappeared, of 84 shortened, of 42 not changed, and of 3 enlarged. Follow-up of endocrine symptoms suggested that endocrine of 50 cases was abnormal, of 29 recovery to normal value, and of 19 not changed obviously. ② Second gamma-knife treatment: At 6- 24 months after gamma-knife treatment, 15 patients with pituitary tumor received second gamma-knife treatment; especially, one patient received for the three times. The conditions in details were recorded as follows: The peri-dose for the first treatment ranged from 8 to 12 Gy; in addition, that for the second one ranged from 6 to 12 Gy. ③ Postoperative complications: The complications were as follow: hypopituitarism occurred in 2 patients (0.7%); tumor apoplexy in 2 patients (0.7%);weakened eyesight in 5 patients (1.8%); 3 of which recovered with dehydration and hormone treatment; 2 patients received tumor resection and optical nerve decompression operation with dehydration and hormone treatment ineffectively.CONCLUSION: Gamma-knife is effective and safe for pituitary adenoma.
基金the National Natural Science Foundation of China, No. 30672239
文摘BACKGROUND: Induced differentiation strategies and cytochemical properties of human embryonic stem ceils (hESCs) have been investigated. However, the electrophysiological functions of tyrosine hydroxylase (TH)-positive cells dedved from hESCs remain unclear. OBJECTIVE: To investigate the differentiation efficiency of TH-positive cells from hESCs in vitro using modified four-step culture methods, including embryoid body formation, and to examine the functional characteristics of the differentiated TH-positive cells using electrophysiological techniques. DESIGN, TIME AND SETTING: Neuroelectrophysiology was performed at the Reproductive Medicine Center and Stem Cell Research Center, Peking University Third Hospital, and the Neuroscience Research Institute and Department of Neurobiology, Peking University, from September 2004 to August 2008. MATERIALS: The hESC line, PKU-1.1, a monoclonal cell line derived from a pre-implantation human blastocyst in the Reproductive Medical Center of Peking University Third Hospital. The patch clamp recording system was provided by the Neuroscience Research Institute and Department of Neurobiology, Peking University. METHODS: The hESC line was induced to differentiate into TH-positive cells in vitro using a modified four-step culture method, including the formation of embryoid body, as well as the presence of sonic hedgehog and fibroblast growth factor 8. The cell karyotype was assessed by G-banding karyotype analysis techniques and specific markers were detected immunocytochemically. Whole-cell configuration was obtained after obtaining a tight seal of over 1 GΩ. Ionic currents were detected by holding the cells at -70 mV and stepping to test voltages between -80 and 40 mV in 10-mV increments in voltage-clamp configuration. MAIN OUTCOME MEASURES: We measured the cell karyotype, specific cell markers, and the electrophysiological properties of the voltage-gated ion channels on the cell membrane of TH-positive dopaminergic cells differentiated from our hESCs line in vitro. RESULTS: The differentiated cells had a consistent appearance, and the majority of cells (〉 90%) expressed TH and β-tubulion, as well as the neural progenitor marker, nestino Cell karyotype analysis demonstrated that all of the hESCs had a stable and normal karyotype (46, XX) after differentiation. In addition, patch clamp recording showed that the 10 recorded TH-positive cells exhibited a fast inward current when the test voltage depolarized to -30 mV, and a delayed outward current when the test voltage depolarized to -10 mV. The peak of inward current was obtained at voltage between 10 mV and 0 mV, while the peak of outward current was obtained at 40 mV. The average peak of inward current density was ( -50.05 ± 15.50) pA/pF, and the average peak of outward current density was (41.98 ± 13.55) pA/pE CONCLUSION: More than 90% of the differentiated hESC-derived cells induced by the modified four-step culture method exhibit dopaminergic neuronal properties, including general electrophysiological functional properties, such as functional potassium and sodium channels.
基金supported by the Youth Science and Technology Innovation Special Foundation of Xinjiang Production and Construction Corps, China, No. 2010JC33
文摘The γ-aminobutyric acid neurotransmitter in the spinal cord dorsal horn plays an important role in pain modulation through primary afferent-mediated presynaptic inhibition. The weakening of γ-aminobutyric acid-mediated presynaptic inhibition may be an important cause of neuropathic pain. γ-aminobutyric acid-mediated presynaptic inhibition is related to the current strength of γ-aminobutyric acid A receptor activation. In view of this, the whole-cell patch-clamp technique was used here to record the change in muscimol activated current of dorsal root ganglion neurons in a chronic constriction injury model. Results found that damage in rat dorsal root ganglion neurons following application of muscimol caused concentration-dependent activation of current, and compared with the sham group, its current strength and γ-aminobutyric acid A receptor protein expression decreased. Immunofluorescence revealed that γ-aminobutyric acid type A receptor α2 subunit protein expression decreased and was most obvious at 12 and 15 days after modeling. Our experimental findings confirmed that the y-aminobutyric acid type A receptor α2 subunit in the chronic constriction injury model rat dorsal root ganglion was downregulated, which may be one of the reasons for the reduction of injury in dorsal root ganglion neurons following muscimol-activated currents.
文摘Using whole-cell patch-clamp technique, the present study examined the effects of captopril on membrane currents of ventricular myocytes. The results were as follows: when the concentration of captopril was 6×10-4 mol, the fast inward sodium current and slow inward calcium current were all decreased significantly (P<0.05). Besides, when the concentration exceeds 10-3 mol,the delayed rectifier potassium current was increased (P<0. 05) as well. But if the experimental concentration was under 6×10-4 mol, these three kinds of currents was not affected. These findings suggested that captopril has no direct antiarrhythmic effect at the average therapeutic serum level.
基金supported by a grant from the National Natural Sciences Foundation of China (No. 30671743)
文摘The roles of voltage-dependent K^+ channels during activation and damage in alveolar macrophages (AMs) exposed to different silica particles were examined. Rat AMs were collected by means of bronchoalveolar lavage, and were adjusted to 5× 10^5/mL. After AMs were exposed to different concentrations (0, 25, 50, 100, 200 μg/mL) of quartz particles and 100 μg/mL amorphous silica particles for 24 h, the voltage-depended K^+ current in AMs was measured by using patch clamp technique. Meanwhile the leakage of lactate dehydrogenase (LDH) and the viability of AMs were detected respectively. Patch clamp studies demonstrated that AMs possessed outward delayed and inward rectifying K^+ current. Exposure to quartz particles increased the outward delayed K^+ current but it had no effect on inward rectifier K^+ current in AMs. Neither of the two K^+ channels in AMs was affected by amorphous silica particles. Cytotoxicity test showed that both silica particles could damage AM membrane and result in significant leakage of LDH (P〈0.05). MTT studies, however, showed that only quartz particles reduced viability of AMs (P〈0.05). It is concluded that quartz parti- cles can activate the outward delayed K^+ channel in AMs, which may act as an activating signal in AMs to initiate an inflammatory response during damage and necrosis in AMs induced by exposure to quartz particle. K^+ channels do not contribute to the membrane damage of AMs.
基金supported by the National Natural Science Foundation of Guangdong (2020A1515010777)the Fund of Chinese Medicine Bureau of Guangdong Province (20201142)。
文摘Objective: Heart failure(HF), a worldwide health condition, is the result of many cardiovascular diseases.The traditional Chinese medicine(TCM) Xiaoyu Jiangzhi capsule(XYC) has long been in use in China to treat hyperlipidemia and inhibit platelet aggregation. This study explores the effects of XYC on heart failure(HF) and its detailed mechanisms.Methods: Isoproterenol(ISO, 30 mg/kg) was injected intraperitoneally for 7 days to copy a HF model of 10-12 weeks old, 20-30 g male mice. We then compared the CON(control) group, ISO(HF model)group, MET(metoprolol) group, and XYC group. Cardiac systolic function and left wall thickness were evaluated by echocardiograph. Using western blot analysis, we detected the proteins of calmodulin dependent protein kinase Ⅱ(Ca MKII) and sarco/endoplasmic reticulum Ca^(2+)-ATPase(Serca). Furthermore, ts A201 cells were cultured and the human CaV1.2 calcium channel current(hCaV1.2) were detected by patch clamp experiments.Results: XYC reduced HF, inhibiting the protein expression of Ca MKII, but Serca did not change significantly. Moreover, XYC inhibited the peak amplitude of the hCaV1.2 current, depolarizing shifted the activation curve 27.6 mV, and shifted the inactivation curve toward a positive potential 17.6 mV. The fraction recovered from inaction was reduced in XYC group compared with that in CON group.Conclusion: XYC could inhibit ISO-induced HF by reducing the Ca^(2+)/Ca MKII signaling pathway in mice.
基金supported by the National Science Foundation of China (Grant No. 81000456)the Science and Technology Department of Sichuan Province (Grant No. 2009SZ0171)
文摘Chlorogenic acid(5-caffeoylquinic acid, CGA) is a phenolic compound that is found ubiquitously in plants, fruits and vegetables and is formed via the esterification of caffeic acid and quinic acid. In addition to its notable biological functions against cardiovascular diseases, type-2 diabetes and inflammatory conditions, CGA was recently hypothesized to be an alternative for the treatment of neurological diseases such as Alzheimer's disease and neuropathic pain disorders. However, its mechanism of action is unclear.Voltage-gated potassium channel(Kv) is a crucial factor in the electro-physiological processes of sensory neurons. Kv has also been identified as a potential therapeutic target for inflammation and neuropathic pain disorders. In this study, we analysed the effects of CGA on the two main subtypes of Kv in trigeminal ganglion neurons, namely, the IK,Aand IK,Vchannels. Trigeminal ganglion(TRG)neurons were acutely disassociated from the rat TRG, and two different doses of CGA(0.2 and 1 mmol·L21) were applied to the cells.Whole-cell patch-clamp recordings were performed to observe alterations in the activation and inactivation properties of the IK,Aand IK,Vchannels. The results demonstrated that 0.2 mmol·L21CGA decreased the peak current density of IK,A. Both 0.2 mmol·L21and1 mmol·L21CGA also caused a significant reduction in the activation and inactivation thresholds of IK,Aand IK,V. CGA exhibited a strong effect on the activation and inactivation velocities of IK,Aand IK,V. These findings provide novel evidence explaining the biological effects of CGA, especially regarding its neurological effects.
文摘The study examined the inhibitory effect of Atractylodes macrocephala (AM) on the uterine contraction during premature delivery and explored its electrophysiological mechanism by studying the effects of AM on the Ca^2+-activated K^+ currents of pregnant human myometrial smooth muscle cells with or without the treatment with intedeukin-6. Single cells were acutely isolated from pregnant human myometrial smooth muscles. Whole-cell Ca^2+-activated K^+ currents were recorded by using an Axopatchl-D amplifier. The cells were divided into three groups: group A in which AM was added into perfusate, group B, in which interleukin-6 was added into perfusate) and group C in which AM was added into perfusate after addition of interleukin-6. IL-6 10 ng/mL inhibited BKca by 36.9%±13.7% as compared with control (P〈0.01). AM at 2 mg/mL raised BKca by 36.7%±22.6% or 45.2%±13.7% with or without the treatment of IL-6, respectively (P〈0.01). It is concluded that AM was able to enhance the BKca of pregnant human myometrial smooth muscle cells treated or untreated with interleukin-6 and its effect on the BKca IL-treated cells was stronger that its effect on BKca of untreated cells. Our results suggested that AM can help to maintain the membrane potentials and the resting status of pregnant human myometrial smooth muscle cells.
基金This project was supported by a grant from the National Natural Science Foundation of China !(No. 39330210).
文摘Summary: To investigate the exact mechanism of epileptogenesis induced by coriaria lactone (CL), the effect of CL on NMDA receptor mediated current (IAsp) in rat hippocampal CA1 neu- rons was investigated by using nystatin perforated whole-cell patch clamp. 10-6-10-4 mol/L Asp acted on NMDA receptors and elicited an inward current (IAsp) at a holding potential (VH) of -40 mV in presence of 10-6 mol/L glycine and absence of Mg2+ extracellularly. CL enhanced NMDA receptor mediated current induced by Asp, but had no effect on threshold concentration, EC50, Hill coefficient as well as maximal-effect concentration and reversal potential of IAsp. The effect had no relationship with holding potential. These results showed that CL could enhance NMDA receptor mediated current to increase [Ca2+]i of neurons by acting on Gly site, thereby inducing epilepsy.
基金This work was supported by the National Natural Science Foundation of China(No.30700747)
文摘Objective:To study the effects of sodium ferulate on the ultrarapid delayed rectifier K^+ current(IKur) in human atrial myocytes. Methods:Human atrial myocytes were isolated by enzyme dispersion method. IKur, in human atrial myocytes were recorded by using the whole cell patch clamp. The changes of IKur were compared in the absence and the presence of sodium ferulate. Results:There was no effect of 0.4 g/L sodium ferulate on I-V relation of IKur. However, 0.4 g/L sodium ferulate inhibited IKur to some degrees at each test pulse. The current densities of IKur at +60 mV decreased from 4.997 ± 0.35 PA/PF to 3.331 ± 0.26 PA/PF(n = 6, P 〈 0.05). The inhibitory effect was concentration-dependent. IC50 was(0.41 ±0.03)g/L and the Hill coefficient was 0.95 ± 0.05. Conclusion:Sodium ferulate as a potassium channel blocker can inhibit IKur in human atrial myocytes effectively.
基金Department of Health of Heilongjiang Province, No. 2003-097Youth Science Foundation of Medical Basic Subjects of Harbin Medical University,No. 060035
文摘BACKGROUND: Some experiments have demonstrated that injecting orphanin FQ (OFQ) into lateral ventricle, which can obviously decrease the pain threshold. It is indicated that OFQ is an anti-opiate substance. However, whether OFQ has effects on sensory neuron ion channel in cerebral cortex needs to be further studied. OBJECTIVE: To investigate the effects of OFQ, morphine or their combination on sodium channel current of somatosensory neurons in rat cerebral cortex. DESIGN: Repeated measurement trial. SETTING: Department of Physiology, Harbin Medical University. MATERIALS: Fifty healthy Wistar rats, aged 12 - 16 days, of either gender, were provided by the Experimental Animal Center, Second Hospital Affiliated to Harbin Medical University. OFQ was purchased from Sigma-Aldrich Company, and morphine was provided by the Shenyang First Pharmaceutical Factory. PC2C patch clamp amplifier and LabmasterTLlwere purchased from Yibo Life Science Instrument Co.,Ltd. of Huazhong University of Science and Techgnology. METHODS: This experiment was carried out in the Department of Physiology (provincial laboratory), Harbin Medical University between January 2005 and May 2006. Cortical neurons were acutely isolated from rats, and prepared into cell suspension following culture. ①Sodium channel current of somatosensory neurons in rat cerebral cortex was recorded before and after administration by whole-cell Patch clamp technique after 50 nmol/L OFQ being added to extracellular fluid. ②The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex was recorded before and after administration by the same method after 20 μmol/L morphine being added to extracellular fluid, and then the change of sodium channel current was recorded after 50 nmol/L OFQ being added. MAIN OUTCOME MEASURES: The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex following the administration of OFQ, morphine separately or their combination.. RESULTS: ①The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex was significantly lower after administration of 50 nmol/L OFQ than before at the clampe of the voltage of -30 mV (P 〈 0.05).②The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex was significantly lower after administration of 20 μmol/L morphine than before at the clampe of the voltage of-30 mV (P 〈 0.05). The sodium channel current recovered to - (2 345.24±174.18) pA after 50 nmol/L OFQ was administrated. There were significant differences in the amplitude of Na^+ channel current between two interventions (P 〈 0.05). CONCLUSION: Morphine and OFQ can respectively reduce the amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex, and OFQ can reverse the effect of morphine partly. It is indicated that OFQ can produce antiopioid activity in the central nervous system by influencing sodium channel current.
基金the National Natural Science Foundation of China, No. 30970930
文摘Cannabinoid and nicotinic acetylcholine receptors are strongly associated with algesia. Previous studies in our laboratory have reported inhibitory effects of synthetic cannabinoid WIN55, 212-2 on nicotine-activated currents (Inic), but the underlying mechanisms remain poorly understood. The present study used whole-cell patch clamp techniques to investigate the modulatory effects of synthetic cannabinoid WIN55, 212-2 on Inic in cultured rat trigeminal ganglion neurons. The results revealed several major findings: WIN55, 212-2 inhibited Inic in rat trigeminal ganglion neurons. In addition, when WIN55, 212-2 (3 μmol/L) was applied simultaneously with nicotine (100 μmol/L), the inhibition of WIN55, 212-2 on Inic was reversible, concentration-dependent and voltage-independent This effect was not mediated by CB1, CB2 or VR1 receptors; neither the selective CB1 receptor antagonist AM281, CB2 receptor antagonist AM630 nor VR1 receptor antagonist capsazepine reduced the inhibitory effect of WIN55, 212-2. Further, the inhibition of nicotinic responses by WIN55, 212-2 was not sensitive to the membrane permeable cyclic adenosine monophosphate (cAMP) analog 8-Br-cAMP. The G-protein inhibitor GDP-I3-S (1 mmol/L) did not block the inhibitory effects of WIN55, 212-2 on/n^c, excluding the involvement of G-protein mediation. The results suggested that WIN55, 212-2 inhibits/n^o directly via the neuronal nicotinic acetylcholine receptor, and that this inhibition is non-competitive. WIN55, 212-2 did not act as an open channel blocker of the neuronal nicotinic acetylcholine receptor, and did not affect the desensitization of Into. The results suggest that nicotine receptors may be physically plugged from outside the membrane by drugs containing WIN55, 212-2.
基金The project supported by Innovation Program of Shanghai Municipal Education Commission(15ZZ063)by Research Project of Putuo Hospital,Shanghai University of Traditional Chinese Medicine(2014YJ002)
文摘OBJECTIVE Toad venom(Venenum Bufonis)isalways used for analgesia in China from ancient to modern times,but the effective component of it remains unclear.METHODS In the present study,we investigated the anti-nociceptive effect and the underlying mechanism ofbufalin,an active ingredient fromtoad venom by animal behavior,patch clamp and calcium imaging.RESULTS Bufalin could significantly relieve formalin-induced spontaneous flinching and licking response as well as carrageenan-induced mechanical and thermal hyperalgesia.Using the whole-cel patch-clamp recording,bufalincaused remarkable suppressive effect on the peak currents of Na+channels in dorsal root ganglion neuroblastoma ND7-23 cel line in a U-shaped dependent manner.In addition,bufalinprompted the voltage-dependent activationand caused a negative shift of the fast-state inactivation of Na+channels.However,bufalin produced insignificant effect not onlyon voltage-dependent Kv4.2,Kv4.3 and BK channels,but also on the capsaicin induced Ca2+influx.CONCLUSION The present results indicate bufalin is capable of producing remarkable anti-nociceptive effects whichis probably ascribed to its specific modulation of voltage-gated Na+channels.
文摘The current difference between male and female rabbit ventricular myocytes was investigated for elucidating the mechanism of longer QT interval and higher incidence of drug-associated torsade de pointes in female rabbits than in male rabbits. Whole cell patch clamp technique was used to record APD, I_to, I_K,tail, I_K1 and I_Ca,L of myocytes from left ventricular apex. There was no difference in the membrane capacitance between male and female rabbit myocytes. APD_90 was longer in female rabbits (560.4±26.5 ms, n=15) than in male ones (489.0±20.7 ms, n=14), P<0.05. In female rabbit myocytes, I_K,tail, I_to, I_K1 and I_Ca,L were 0.71±0.05 pA/pF (n=17), 8.28±1.03 pA/pF (n=18), 24.5±3.6 pA/pF (n=12) and 9.0±2.3 pA/pF (n=15) respectively. In male rabbit myocytes, they were 0.84±0.07 pA/pF (n=18), 8.60±1.20 pA/pF (n=18), 25.9±4.5 pA/pF (n=14) and 9.3±2.6 pA/pF (n=16) respectively. I_K,tail in female rabbits was significantly lower than that of male rabbits (P<0.05), but there was no difference in I_to, I_K1 and I_Ca,L between male rabbits and female rabbits (P>0.05). The lower I_K,tail of female rabbit myocytes may contribute to the longer repolarization and the higher incidence of drug-associated torsade de pointes.