Acupuncture inhibits Notch1 and Hes1 protein expression in the basal ganglia of rats with cerebral hemorrhage

Notch pathway activation maintains neural stem cells in a proliferating state and increases nerve repair capacity. To date, studies have rarely focused on changes or damage to signal transduc- tion pathways during cerebral hemorrhage. Here, we examined the effect of acupuncture in a rat model of cerebral hemorrhage. We examined four groups： in the control group, rats received no treatment. In the model group, cerebral hemorrhage models were established by infusing non-hep-arinized blood into the Brain. In the acupuncture group, modeled rats had Baihui （DU20） and Qubin （GBT） acupoints treated once a day for 30 minutes. In the DAPT group, modeled rats had 0.15 μg/mL DAPT solution （10 mL） infused into the brain. Immunohistochemistry and western blot results showed that acupuncture effectively inhibits Notch 1 and Hesl protein expression in rat basal ganglia. These inhibitory effects were identical to DAPT, a Notch signaling pathway inhibitor. Our results suggest that acupuncture has a neuroprotective effect on cerebral hemorrhage by in- hibiting Notch-Hes signaling pathway transduction in rat basal ganglia after cerebral hemorrhage.

Protective effect of electroacupuncture and moxibustion on gastric mucosal damage and its relation with nitric oxide in rats

INTRODUCTION Gastric mucosal injury is one of the common disorders,there are many reports subjicted to its pathogenesis treatment and prevention.We investigated the protective effect

Protective Action of Acupuncture and Moxibustion on Gastric Mucosa in Model Rats with Chronic Atrophic Gastritis

Objective: To probe the mechanism of acupuncture and moxibustion in atrophic gastritis so as to provide a basis for clinical treatment. Method: Observe the effects of acupuncture and moxibustion at the points of Zusanli, Zhongwan and Tianshu on gastric mucosa in model rats with chronic atrophic gastritis. Results:Acupuncture and moxibustion can increase the contents of PGE2α, PGF2α and cAMP, and decrease the content of cGMP in the tissue of gastric mucosa. Conclusion: Acupuncture and moxibustion shows cytoprotection on gastric mucosa, so it is an effective method for treating chronic atrophic gastritis.

Long-term acupuncture treatment has a multitargeting regulation on multiple brain regions in rats with Alzheimer＇s disease：a positron emission tomography study

The acute effect of acupuncture on Alzheimer＇s disease,i.e.,on brain activation during treatment,has been reported.However,the effect of long-term acupuncture on brain activation in Alzheimer＇s disease is unclear.Therefore,in this study,we performed long-term needling at Zusanli（ST36）or a sham point（1.5 mm lateral to ST36）in a rat Alzheimer＇s disease model,for 30 minutes,once per day,for 30 days.The rats underwent 18F-fluorodeoxyglucose positron emission tomography scanning.Positron emission tomography images were processed with SPM2.The brain areas activated after needling at ST36 included the left hippocampus,the left orbital cortex,the left infralimbic cortex,the left olfactory cortex,the left cerebellum and the left pons.In the sham-point group,the activated regions were similar to those in the ST36 group.However,the ST36 group showed greater activation in the cerebellum and pons than the sham-point group.These findings suggest that long-term acupuncture treatment has targeted regulatory effects on multiple brain regions in rats with Alzheimer＇s disease.

Comparison of the analgesic effects between electro-acupuncture and moxibustion with visceral hypersensitivity rats in irritable bowel syndrome

AIM To observe whether there are differences in the effects of electro-acupuncture(EA) and moxibustion(Mox) in rats with visceral hypersensitivity. METHODS EA at 1 m A and 3 m A and Mox at 43?℃ and 46?℃ were applied to the Shangjuxu(ST37, bilateral) acupoints in model rats with visceral hypersensitivity. Responses of wide dynamic range neurons in dorsal horns of the spinal cord were observed through the extracellular recordings. Mast cells(MC) activity in the colons of rats were assessed, and 5-hydroxytryptamine(5-HT), 5-hydroxytryptamine 3 receptor(5-HT3R) and 5-HT4Rexpressions in the colons were measured.RESULTS Compared with normal control group, responses of wide dynamic range neurons in the dorsal horn of the spinal cord were increased in the EA at 1 m A and 3 m A groups(1 m A: 0.84 ± 0.74 vs 2.73 ± 0.65, P < 0.001; 3 m A: 1.91 ± 1.48 vs 6.44 ± 1.26, P < 0.001) and Mox at 43?℃ and 46?℃ groups(43?℃: 1.76 ± 0.81 vs 4.14 ± 1.83, P = 0.001; 46?℃: 5.19 ± 2.03 vs 7.91 ± 2.27, P = 0.01). MC degranulation rates and the expression of 5-HT, 5-HT3 R and 5-HT4 R in the colon of Mox 46?℃ group were decreased compared with model group(MC degranulation rates: 0.47 ± 0.56 vs 0.28 ± 0.78, P < 0.001; 5-HT: 1.42 ± 0.65 vs 7.38 ± 1.12, P < 0.001; 5-HT3R: 6.62 ± 0.77 vs 2.86 ± 0.88, P < 0.001; 5-HT4R: 4.62 ± 0.65 vs 2.22 ± 0.97, P < 0.001). CONCLUSION The analgesic effects of Mox at 46?℃ are greater than those of Mox at 43?℃, EA 1 m A and EA 3 m A.

Effects of cluster needling of scalp acupuncture on neurofilament protein 200 and signal transducer and activator of transcription 3 in rats with acute cerebral ischemia

Objective:Cluster needling at scalp acupoints has showed satisfying effects with acute cerebral ischemia in clinic whereas the mechanisms are not yet clear completely.This study investigated the influence of cluster needling at scalp acupoints on neurological function,as well as on neurofilament protein 200(NF200)and signal transducer and activator of transcription 3(STAT3)expression,in rats with acute cerebral ischemia.Methods:Fifty-four Sprague-Dawley rats were randomly assigned in equal numbers to the false operation(group F),model(group M),or cluster needling scalp acupuncture(group C)groups.Each group was divided into three subgroups,of six rats each,by acupuncture treatment time(24 h,7 days,and 14 days).The rat local cerebral ischemia model was prepared using a modified suture occlusion method.Group C rats were treated by cluster needling scalp acupuncture,while groups F and M did not receive acupuncture treatment.Neurological effects were evaluated using the Longa score.NF200 and STAT3 expression were measured by western blotting.Results:At 24 h,there were no statistical difference between group C and group M in nerve function(P>.05).On days 7 and 14,nerve function scores in group C were significantly lower than that in group M(respectively were P<.05 and P<.01).In addition,on days 14,expression of NF200 was significantly higher in group C compared with group M(P<.05).Compared with group M,STAT3 expression was also higher in group C on days 7 and 14,although these differences were not statistically significant(both P>.05).Conclusion:Cluster needling scalp acupuncture were efficient in improving nerve function scores in rats with cerebral ischemia,and promoting the recovery of motor function.These improvements were associated with increases in NF200 and STAT3 expression.

Effects of acupuncture and moxibustion on chromatin remodeling-related enzymes in the colon tissue of rats with Crohn disease

Objective:To investigate the regulatory effects of acupuncture and moxibustion on chromatin remodeling complex core catalytic subunit of Brahma-related gene 1(Brg1),histone deacetylase(HDAC)3,HDAC9,and males absent on the first(MOF)in the colon tissue of rats with Crohn disease(CD).Methods:Using the random number table method,60 male Sprague-Dawley rats were divided into 5 groups,including a normal group,a model group,an acupuncture group,a medicinal cake-insulated moxibustion group,and an acupuncture-moxibustion group,with 12 rats in each group.CD rat models were prepared using 2,4,6-trinitrobenzene sulfonic acid(TNBS)in all groups except the normal group.The normal and model groups received no interventions.In the acupuncture group,rats were intervened with acupuncture at bilateral Zusanli(ST36)and Shangjuxu(ST37),20 min/session,once a day.The medicinal cake-insulated moxibustion group received medicinal cake-insulated moxibustion at Qihai(CV6)and bilateral Tianshu(ST25)with 2 cones per point per session,once a day.The acupuncture-moxibustion group received both acupuncture and moxibustion interventions simultaneously.Each intervention was performed for 10 consecutive days.Observations included general condition,disease activity,macroscopic damage,and pathological changes in the rat’s colon tissue.Realtime fluorescence quantitative polymerase chain reaction was used to measure the mRNA expression of chromatin remodeling-related enzymes Brg1,HDAC3,and HDAC9,while Western blotting detected the protein expression of Brg1,HDAC3,HDAC9,and MOF in rat’s colon tissue.Results:The model group showed significantly increased diarrhea score,occult blood score,macroscopic damage score of colon tissue,and colon macroscopic damage index(CMDI)score,as well as elevated mRNA expression levels of HDAC3 and HDAC9,protein expression levels of HDAC3,HDAC9,and MOF,and decreased mRNA and protein expression levels of Brg1 compared to the normal group(P<0.01).In contrast,compared to the model group,the diarrhea score,occult blood score,macroscopic damage score,CMDI score,mRNA expression levels of HDAC3 and HDAC9,and protein expression levels of HDAC3,HDAC9,and MOF were significantly reduced(P<0.05 or P<0.01),while the mRNA and protein expression levels of Brg1 were significantly increased(P<0.01)in the acupuncture group,the medicinal cake-insulated moxibustion group,and the acupuncture-moxibustion group.Conclusion:Both medicinal cake-insulated moxibustion and acupuncture,either used alone or in combination,can regulate the abnormal expression of chromatin remodeling-related enzymes Brg1,HDAC3,HDAC9,and MOF in the colon tissue,thus reducing colon inflammation in CD rats.

Effects of acupuncturing Tsusanli (S_T36) on expression of nitric oxide synthase in hypothalamus and adrenal gland in rats with cold stress ulcer

AIM： To study the protective effect of acupuncturing Tsusanli （ST36） on cold stress ulcer, and the expression of nitric oxide synthase （NOS） in hypothalarnus and adrenal gland. METHODS： Ulcer index in rats and RT-PCR were used to study the protective effect of acupuncture on cold stress ulcer, and the expression of NOS in hypothalamus and adrenal gland. Images were analyzed with semi-quantitative method. RESULTS： The ulcer index significantly decreased in rats with stress ulcer. Plasma cortisol concentration was up regulated during cold stress, which could be depressed by pre-acupuncture. The expression of NOS1 in hypothallamus increased after acupuncture. The increased expression of NOS2 was related with stress ulcer, which could be decreased by acupuncture. The expression of NOS3 in hypothalamus was similar to NOS2, but the effect of acupuncture was limited. The expression of NOS2 and NOS3 in adrenal gland increased after cold stress, only the expression of NOS1 could be repressed with acupuncture. There was no NOS2 expression in adrenal gland in rats with stress ulcer. CONCLUSION： The protective effect of acupuncturing Tsusanli （ST36） on the expression of NOS in hypothalamus and adrenal gland can be achieved.

Effects of preventive acupuncture or moxibustion at the Guanyuan acupoint on expression of interleukin-2 and its receptors in hypothalamus, serum, and uterus of menopausal rats

BACKGROUND： Previous studies have indicated preventive acupuncture and moxibustion at the Guanyuan point have positive effects on the immune system of menopausal rats. OBJECTIVE： To observe the effects of preventive acupuncture or moxibustion at the Guanyuan point on interleukin-2 （IL-2） and its receptor （IL-2R mRNA） levels in hypothalamus, serum, and uterus of menopausal rats. DESIGN： Randomized control animal experiment. SETTING： School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine. MATERIALS： This study was performed at the Laboratory of Beijing University of Chinese Medicine from November 2004 to July 2005. The vaginal smear method was used to determine the estrus cycle of 9.5-month-old Sprague Dawley （SD） female rats. Three continuous estrus cycles were measured, and 160 rats with estrus cycle disorder, weighing （300 _＋ 20） g, were included in this study. The rats were randomly divided into four groups： preventive acupuncture group （n =48）, preventive moxibustion group （n =48）, model groups （n =48）, and 10-month-old group （n =16）. Sixteen 3.5-month-old healthy SD female rats, weighing （240 ± 20） g, were also assigned to the young group. All rats were clean grade and were purchased from the Laboratory Animal Center, Beijing University of Chinese Medicine. The experiment was in accordance with animal ethics standards. METHODS： Acupuncture was applied to the preventive acupuncture group at 10 months of age. The needle was inserted upward and perpendicularly two fen to a point that was two fen below the Guanyuan point. The rats freely moved around during needle retention （20 minutes）. Acupuncture was administered twice a week and continued for eight weeks. A moxa cone （small） moxibustion was applied to the preventive moxibustion group at 10 months of age. The rats were caught gently and kept at the same position as the preventive acupuncture group. The moxa cone was ignited and held to the Guanyuan point directly. Only one moxa cone was used each time. After moxibustion, the local skin turned slightly red. Moxibustion was used twice a week and continued for eight weeks. Nothing except gently catching and lying was administered to the model group and the young group. MAIN OUTCOME MEASURES： Radioimmunoassay, in situ hybridization, and similar methods were employed to detect IL-2 in serum and the uterus, as well as IL-2R mRNA expression in the hypothalamus and uterus arcuate nuclei, of the preventive acupuncture group, preventive moxibustion group, model groups （at 12, 14, and 16 months）, 10 month old group （at 10 months）, and the young group （at 4 months）. RESULTS： All 176 rats were included in the final analysis. （1） IL-2R mRNA： IL-2R mRNA expression decreased significantly in the uterus of 12-, 14-, and 16-month-old model groups and in the hypothalamus of 14- and 16-month-old model groups （P 〈 0.05-0.01）. Compared to the same month-aged model groups, the expression of IL-2R mRNA increased significantly in the hypothalamus and uterus of 16-month-old preventive acupuncture or moxibustion groups （P 〈 0.05-0.01）.（2） IL-2：IL-2 in serum of 12- and 14-month-old model groups and in the uterus of 12-, 14-, and 16-month-old model groups decreased significantly compared to the young group （P 〈 0.05-0.01）. IL-2 serum levels increased significantly in the 16-month-old group with preventive moxibustion; in addition, IL-2 expression levels increased significantly in the uterus of the 12-month-old group with preventive moxibustion, as well as the 12- and 16-month-old groups with preventive acupuncture （P 〈 0.05-0.01）. CONCLUSION： As a rat ages and becomes menopausal, IL-2R mRNA expression in the hypothalamus and uterus, as well as IL-2 levels in the serum and uterus, decrease. However, preventive acupuncture and preventive moxibustion can repress the decrease to some extent. This effect is even greater in the late stages of menopause （16-month-old rats）.

Study on Action of Acupuncture on Ventromedial Nucleus of Hypothalamus in Obese Rats

Effects of acupuncture on the spontaneous discharge of nerve cells and levels of monoamine neurotransmitters in ventromedial nucleus of hypothalamus (VMH) of the obese rats were investigated. Results indicated that the levels of tyrosine (Tyr) and dopamine (DA) were lowered, the levels of 5-hydroxytryptamine (5-HT) and 5-hydoxyindole acetic acid (5-HIAA) increased, and the frequency of the spontaneous discharge of nerve cells in VMH lowered in the obese rat group as compared with the normal group. When acupuncture obtained the effect of weight reduction, the frequency of spontaneous discharges of nerve cells in VMH were markedly increased, and the levels of Tyr, DA and tryptamine (Typ) and 5-HT/5-HIAA ratio elevated, and the 5-HT level lowered. It is suggested that the virtuous regulative action of acupuncture on VMH might be one of the key factors in acupuncture for weight reduction.

Inhibitory effect of acupuncture on neuronalapoptosis in rats after cerebral ischemia

BACKGROUND： Delayed neuronal death after total cerebral ischemia may accompany with apoptosis, but acupuncture may play a certain role in protecting nerve through inhibiting ischemic neuronal apoptosis. OBJECTIVE： To observe the effect of acupuncture on neuronal apoptosis in rats after cerebral ischemia and analyze its cerebral protective mechanism. DESIGN： Contrast observation among groups. SETTING： Heilongjiang University of Traditional Chinese Medicine. MATERIALS： A total of 30 male healthy Wistar rats of general grade and weighing （250±20） g were randomly divided into three groups, including sham operation group, cerebral ischemia group and acupuncture group with 10 rats in each group. Apoptosis in situ kit was provided by Baolingman Company, Germany. METHODS： The experiment was carried out in the Laboratory Center, Heilongjiang University of Traditional Chinese Medicine from May to November 2004. ① Rats in the cerebral ischemia group and the acupuncture group were used to establish total cerebral ischemic models with four vessels occlusion; in addition, models in the sham operation group were established with the same method as mentioned above. However, four vessels of rats in the sham operation were exposured and cerebral ischemia did not occur. Rats in the acupuncture group were given acupuncture treatment after operation. Needle of 40 mm in length was used to acupuncture bilateral Zusanli （St 36） and Quchi （LI 11） with the depth of 3 mm, and then bilateral acupoints were connected with KWD-808Ⅱ omnipotenc impulse electro-therapeutic apparatus （frequency： 1 Hz; thin waves; voltage： 2 V） once a day for totally 30 minutes. Meanwhile, needle of 25 mm in length was used to acupuncture Baihui （Du 20） with the depth of 3 mm, and then the needle was twirled once every 5 minutes for 30 minutes in total. The course was 7 days. ② Neuronal injuries in hippocampal CAI area after cerebral ischemia were observed with Nissl body staining method at 7 days after treatment; neuronal apoptosis was observed with TUNEL staining; manifestations of neuronal apoptosis in cerebral cortex and hippocampal CAI area were observed with electron microscope. MAIN OUTCOME MEASURES： Neuronal injuries in hippocampal CAI area after cerebral ischemia; neuronal apoptosis in cerebral cortex and hippocampal CAI area after cerebral ischemia; morphological changes under electron microscope. RESULTS： Among 30 Wistar rats, 24 rats were involved in the final analysis. ① Expression of positive neurons in cerebral cortex and hippocampal CAI area with Nissl body staining： Neuronal defect was obvious in cerebral cortex and hippocampal CAI area in the cerebral ischemia group as compared with that in the sham operation group （P 〈 0.05）, and neuronal defect was decreased in hippocampal CAI area in the cerebral ischemia group as compared with that in the acupuncture group （P 〈 0.05）. ② Expression of positive neurons in cerebral cortex and hippocampal CAI area with TUNEL staining： Positive neurons with TUNEL staining were not observed in the sham operation group, but positive neurons were increased in the cerebral ischemia group as compared with those in the acupuncture group （P 〈0.05）. ③ Observational results of electron microscope： Neuronal apoptosis was not found in the sham operation group; neuronal apoptosis was rarely found in the acupuncture group; neuronal apoptosis was typical in the cerebral ischemia group. CONCLUSION： Delayed neuronal death after total cerebral ischemia may accompany with apoptosis, but acupuncture may play a certain role in protecting nerve through inhibiting ischemic neuronal apoptosis.

Electroacupuncture-regulated neurotrophic factor mRNA expression in the substantia nigra of Parkinson's disease rats

Acupuncture for the treatment of Parkinson＇s disease has a precise clinical outcome. This study investigated the effect of electroacupuncture at Fengfu （GV16） and Taichong （LR3） acupoints in rat models of Parkinson＇s disease induced by subcutaneous injection of rotenone into rat neck and back. Reverse transcription-PCR demonstrated that brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor mRNA expression was significantly increased in the substantia nigra of rat models of Parkinson＇s disease, and that abnormal behavior of rats was significantly improved following electroacupuncture treatment. These results indicated that electroacupuncture treatment upregulated brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor mRNA expression in the substantia nigra of rat models of Parkinson＇s disease. Thus, electroacupuncture may be useful in the treatment of Parkinson＇s disease.

Effects of Acupuncture on Monoamine Neurotransmitters in Raphe Nuclei in Obese Rats

Effects of acupuncture on the levels of neurotransmitters in the raphe nuclei were investigated in obeserats.It was found that the levels of tryptophan (Trp) and 5-hydroxyindoleacetic acid (5-HIAA) wereincreased,and 5-hydroxytryptamine (5-HT) level and 5-HT/5-HIAA ratio decreased in the raphe nucleiof the obese group as compared with the normal group;and that acupuncture could produce weightreduction,increase the 5-HT level and 5-HT/5-HIAA ratio,and decrease the contents of Trp and5-HIAA,but did not change the levels of dopamine (DA) and noradrenaline (NA).It is indicated thatbenign regulative action of acupuncture on 5-HT and its metabolism in the raphe nuclei is possibly oneof the factors for reducing weight by acupuncture.

Acupuncture attenuates the development of diabetie peripheral neuralgia by regulating spinal microglia in rats

Diabetic peripheral neuropathy(DPN)is a chronic microvascular complication of diabetes.However,studies on effect of acupuncture in DPN are rare.The purpose of this study was to research the mechanism of acupuncture in DPN rats.Rats rendered diabetic with a single injection of 35 mg/kg STZ.The STZ-diabetic rats were randomly divided into two groups:DPN model and acupuncture groups.After 14 days treatment,the therapeutic efficacy of acupuncture was assessed using MWT and TWI evaluation of rats.

The change of unit discharges in the dorsomedial nucleus of the hypothalamus and the effect of electroacupuncture on it in trauma rats

The unit discharges in the dorsomedial nucleus of the hypothalamus (DMH) were extracellularly recorded with microelectrodes in trauma models of male Sprague-Dawley rats. The main results were as follows : (1) The total number of spontaneously discharged units in DMH of trauma rats was markedly decreased while the number of pain-sensitive units (nociceptive neurons) did not change obviously. (2) The electroacupuncture (EA)at Zusanli (ST36) and Sanyinjiao (Sp6) decreased the spontaneous and pain-evoked firing (discharge) rate, and shortened the durations of pain-evoked firing of pain-excited units in DMH. It also resulted in increase in spontaneous discharge rate of pain-inhibited units and relief of their inhibition from the peripheral noxious stimulations. These results suggest that the depression of the function of DMH caused by the persistent peripheral noxious stimulation may be one of the mechanisms of hyperalgesia. EA at the points of Zusanli (ST36) and Sanyinjiao (Sp6) can interfere with the conduction of pain signals to DMH in the central nervous system and thereby prevent DMH from being depressed , which may partly account for the central mechanisms of acupuncture-analgesia in chronic pain state.

A hippocampal anti-hypertensive mechanism induced by twirling reinforcing-reducing manipulation in rats

Objective:To investigate a hippocampal anti-hypertensive mechanism induced by twirling reinforcingreducing manipulation(TRRM)using proteomics in rats.Methods:Forty-two male spontaneously hypertensive rats were randomly divided into 3 groups,and 14 Wistar-Kyoto rats were served as control group.In the twirling reinforcing(TRF)group,needles were directly inserted into the Taichong(LR 3)point,then thumbs were moved heavily forward and lightly backward for 3 min,while needles remained inserted for 17 min.In the twirling reduction(TRD)group,the same treatment was applied as in the TRF group except that the thumb moved lightly forward and heavily backward.In the model and control groups,only the corresponding grasping and fixation were applied.All interventions were conducted for 14 days.The blood pressure(BP)of all rats was measured one day before intervention and every other day after.Then hematoxylin-eosin(H&E)staining,label-free and parallel reaction monitoring proteomic techniques were used to assess hippocampal samples from each group.Results:Systolic BPs showed a significant decrease in the TRF and TRD groups compared with the model group(P<.01).In the model group,H&E staining showed obvious pathological changes in the hippocampus,while in the TRF and TRD groups,the hippocampal morphology was only slightly altered.Labelfree proteomic analysis revealed 1163 differential protein expressions between groups.Gene Ontology enrichment analysis confirmed that the differentially expressed proteins were enriched in different biological processes,cellular components,and molecular functions.Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that TRRM proteins were expressed in the serotonergic synapse pathway,renin-angiotensin system,the mitogen-activated protein kinase signaling pathway,and the peroxisome pathway,which were all also related to BP regulation.Conclusion:TRRM can significantly lower the BP of SHRs.The mode of action may be through the activation of various protein pathways in the hippocampus that are related to BP regulation.

基于NLRP3炎症小体介导的细胞焦亡探讨针康法对缺氧缺血性脑损伤新生大鼠的影响

目的:观察针康法对缺氧缺血性脑损伤(HIBD)新生大鼠神经元NLRP3炎症小体及细胞焦亡的影响,探讨其改善缺氧缺血性脑损伤后运动功能缺损的可能机制。方法:选择120只7 d龄大鼠分为正常对照组(A组)、模型对照组(B组)、头穴丛刺组(C组)、康复训练组(D组)和针康组(E组),再均分为24 d、7 d和14 d 3个时间点。结扎颈总动脉构建HIBD动物模型。C组针刺百会以及左右各旁开2 mm处,1次/d,每次留针2 h;D组进行转笼转棒训练,1次/d,10 min/次;E组在头穴丛刺的基础上进行转笼转棒训练。在各时间点,采用网屏实验评定运动功能缺损,TUNEL+Caspase-1免疫荧光双标法观察海马区神经细胞焦亡的情况,蛋白印迹法检测海马区NLRP3、Caspase-1和GSDMD-N端蛋白表达。结果:相较于A组,经缺氧缺血造模处理的4组在各时间点网屏实验评分和海马区神经细胞焦亡率均显著升高,NLRP3、Caspase-1和GSDMD-N端蛋白的表达显著增加,差异具有统计学意义(P<0.05);相比较B组,各治疗组在各时间点网屏实验评分和海马区神经细胞焦亡率均显著减少,NLRP3、Caspase-1和GSDMD-N端蛋白的表达水平降低,其中E组的疗效最明显,差异具有统计学意义(P<0.05)。结论:针康法可改善HIBD新生大鼠的运动功能缺损,且疗效优于单纯针刺或康复疗法,其机制可能与抑制NLRP3炎症小体介导的细胞焦亡有关。

耳甲电针疗法对CUMS诱导抑郁症大鼠的治疗作用及其机制

目的探讨耳甲电针疗法对慢性不可预知温和应激(CUMS)诱导抑郁症大鼠的治疗作用,并基于蛋白激酶A(PKA)/环磷腺苷效应元件结合蛋白(CREB)信号通路探讨其作用机制。方法选择成年雄性SD大鼠24只,随机分为对照组、模型组、氟西汀组、耳甲电针组,每组6只。对照组正常群养,每日灌胃生理盐水,不予特殊干预,连续35天。模型组、氟西汀组、耳甲电针组孤养,均予CUMS暴露35天。氟西汀组实验第14天开始在CUMS暴露前予盐酸氟西汀灌胃,每日1次,连续21天;耳甲电针组实验第14天开始在CUMS暴露前予电针刺激双侧耳甲神门穴,每日1次,每次30 min,连续21天。实验前和实验第14、35天,称量大鼠体质量;通过旷场实验观察大鼠自发活动行为和探索行为;通过糖水偏好实验评估大鼠快感缺失行为。断头取脑,分离海马组织,采用Western blotting法检测海马组织PKA/CREB信号通路蛋白表达。结果与对照组同期比较,模型组实验第14天与第35天体质量、自发活动行为和探索行为以及糖水偏好指数均显著降低(P均<0.05)。与模型组同期比较,氟西汀组和耳甲电针组实验第35天体质量、自发活动行为和探索行为以及糖水偏好指数均显著升高(P均<0.05)。模型组海马组织PKA-α、p-CREB蛋白相对表达量均显著低于对照组(P均<0.05);氟西汀组和耳甲电针组海马组织PKA-α、p-CREB蛋白相对表达量均显著高于模型组(P均<0.05)。结论耳甲电针疗法能够改善CUMS诱导抑郁症大鼠抑郁样行为,其效果与氟西汀相当;激活海马组织PKA/CREB信号通路可能是其作用机制之一。

耳穴贴压联合电针对膝骨关节炎模型大鼠滑膜纤维化及内质网应激的抑制作用研究

目的观察耳穴贴压联合电针对膝骨关节炎模型大鼠滑膜纤维化及内质网应激的抑制作用。方法将60只Sprague-Dawley(SD)大鼠随机分为正常组(10只)和其余组(50只),其余组造模成功后再次随机分为模型组、耳穴组、电针组、联合组及药物组,每组10只。耳穴组使用磁珠贴于大鼠两侧肾耳穴进行干预,电针组予电针干预,联合组予耳穴磁珠贴压联合电针干预,药物组予塞来昔布水溶液灌胃干预。观察各组大鼠膝关节肿胀度的变化。观察各组大鼠血清白介素-1(interleukin-1,IL-1)、白介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、锰超氧化物歧化酶(manganese superoxide dismutase,Mn-SOD)、过氧化氢酶(catalase,CAT)和丙二醛(malondialdehyde,MDA)的水平,观察关节液中环氧合酶-2(cyclooxygenase-2,COX-2)、前列腺素E2(prostaglandin E2,PGE2)和一氧化氮(nitric oxide,NO)的水平,观察各组大鼠滑膜组织中白介素-1β(interleukin-1β,IL-1β)及TNF-α的水平。观察大鼠软骨组织病理学变化,比较各组滑膜组织中葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)、C/EBP同源蛋白(C/EBP-homologous protein,CHOP)、基质金属蛋白酶13(matrix metalloproteinase 13,MMP13)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、金属蛋白酶组织抑制剂1(tissue inhibitor of metalloproteinase 1,TIMP1)、胶原蛋白Ⅰ(collagenⅠ,Col-Ⅰ)、胶原蛋白Ⅱ(collagenⅡ,Col-Ⅱ)mRNA水平和蛋白水平。结果与正常组比较,模型组大鼠关节肿胀度,血清IL-1、IL-6、TNF-α和MDA水平,关节液COX-2、PGE2和NO水平,滑膜组织IL-1β和TNF-α水平,滑膜组织GRP78、CHOP、MMP-13、α-SMA、TIMP1、Col-Ⅰ、Col-ⅡmRNA及蛋白水平,均显著升高(P<0.05);模型组大鼠血清Mn-SOD和CAT水平显著降低(P<0.05)。与模型组比较,耳穴组、电针组、联合组和药物组大鼠关节肿胀度,血清IL-1、IL-6、TNF-α和MDA水平,关节液COX-2、PGE2和NO水平,滑膜组织IL-1β和TNF-α水平,滑膜组织GRP78、CHOP、MMP-13、α-SMA、TIMP1、Col-Ⅰ、Col-ⅡmRNA及蛋白水平,均显著降低(P<0.05);血清Mn-SOD和CAT水平显著升高(P<0.05);大鼠关节软骨组织病理学明显改善。与耳穴组和电针组比较,联合组大鼠关节肿胀度,血清IL-1、IL-6、TNF-α和MDA水平,关节液COX-2、PGE2和NO水平,滑膜组织IL-1β和TNF-α水平,滑膜组织GRP78、CHOP、MMP-13、α-SMA、TIMP1、Col-Ⅰ、Col-ⅡmRNA及蛋白水平,均显著降低(P<0.05);血清Mn-SOD和CAT水平均显著升高(P<0.05)。结论耳穴贴压联合电针能显著改善膝骨关节炎大鼠关节肿胀程度,抑制病理进展,缓解大鼠滑膜组织炎症反应,修复氧化应激损伤,进而修复膝骨关节炎大鼠滑膜组织纤维化及内质网应激反应。

从肝论治取穴针灸通过调控JAK2/STAT3途径对膝骨关节炎大鼠关节软骨的保护作用

目的:探究从肝论治取穴针灸通过调控JAK2/STAT3途径对膝骨关节炎(OA)大鼠关节软骨的保护作用。方法:60只SD大鼠随机分为对照组、OA组、OA+针灸组,每组20只,通过手术构建右膝OA模型。OA+针灸组大鼠接受从肝论治取穴针灸4周。检测软骨退化情况、关节炎症,以及JAK2/STAT3转录和蛋白水平。结果:OA组大鼠OARSI评分高于对照组(P<0.05);干预后,OA+针灸组大鼠OARSI评分低于OA组(P<0.05)。OA组大鼠透明软骨(HC)厚度低于对照组,钙化软骨(CC)厚度高于对照组(P<0.05);干预后,OA+针灸组大鼠HC厚度高于OA组,CC厚度低于OA组(P<0.05)。OA组大鼠白细胞介素-1β(IL-1β)、IL-6、JAK2 mRNA、STAT3 mRNA、JAK2蛋白、STAT3蛋白水平均高于对照组(P<0.05);干预后,OA+针灸组大鼠IL-1β、IL-6、JAK2 mRNA、STAT3 mRNA、JAK2蛋白、STAT3蛋白水平均低于OA组(P<0.05)。结论:从肝论治取穴针灸可通过调控JAK2/STAT3途径发挥对OA大鼠膝关节软骨的保护作用。