Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1genes in pancreatic acinar cells was investigated. Rat panc...Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1genes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10 -3 mol/L, 10 -4 mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10 -3 mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-κB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10 -3 mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10 -5 mol/L atropine) or NF-κB inhibitor (10 -2 mol/L PDTC) could obviously inhibit the activation of NF-κB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-1genes in pancreatic acinar cells in vitro through the activation of NF-κB.展开更多
Objective: To study roles of cholinergic M2 and M3 receptors in the motility reduction of small intestine (SI) in spleen qi deficiency. Methods: 16 male SD rats were randomly divided in the control group and spleen qi...Objective: To study roles of cholinergic M2 and M3 receptors in the motility reduction of small intestine (SI) in spleen qi deficiency. Methods: 16 male SD rats were randomly divided in the control group and spleen qi deficiency group (model group)—8 rats each group;spleen qi deficiency model of the improper diet and overfatigue was established;the SI propelling rate (SIPR) was used to evaluate the SI motility;ELISA was used to measure concentrations of acetylcholine (ACh), cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in the SI tissue;immohistochemistry was employed to detect expressions of cholinergic M2 and M3 receptors. Results: Compared with those in the control group, SIPR was reduced;expression of M2 receptors was increased;and expression of M3 receptors and concentrations of cAMP and PKA were decreased, significantly, in the model group. Conclusions: Expression imbalance of cholinergic M2 and M3 receptors might contribute to the motility reduction of the SI in spleen qi deficiency.展开更多
Idiopathic pulmonary fibrosis is an untreatable lethal lung disease, which is related to the aberrant proliferation of fibroblasts. M<sub>3</sub> muscarinic acetylcholine receptor (M<sub>3</sub>...Idiopathic pulmonary fibrosis is an untreatable lethal lung disease, which is related to the aberrant proliferation of fibroblasts. M<sub>3</sub> muscarinic acetylcholine receptor (M<sub>3</sub>-mAChR) activation exerts proliferative effect on various kinds of cells. However, whether M<sub>3</sub>-mAChR inhibition has a protective effect on pulmonary fibrosis remains unexplored. A rat model of pulmonary fibrosis was established by intratracheal instillation of bleomycin. Darifenacin was used to block M<sub>3</sub>-mAChR. Histological changes were observed using Masson’s Trichrome and hematoxylin and eosin (HE) staining. Hydroxyproline was measured by Hydroxyproline detection kit. Transforming growth factor β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). In vitro, pulmonary fibroblasts were isolated from lungs of neonatal rat. After treatment, the cell viability, Hydroxyproline level was measured by MTT and Hydroxyproline detection kit respectively. The expression level of extracellular signal-regulated kinase (ERK), nuclear factor kappa-B (N-NF-κB), and microRNA-21 (miR-21) was detected by western blot or quantitative real-time PCR (qRT-PCR). Darifenacin relieved the fibrotic effects provoked by bleomycin. The expression level of hydroxyproline, TGF-β1 and TNF-α level was all downregulated after darifenacin treatment. In lung fibroblasts, darifenacin decreased cell viability and hydroxyproline level induced by bleomycin. Besides, phosphorylation-ERK and nuclear N-NF-κB protein level was downregulated, as well as miR-21 level. M<sub>3</sub>-mAChR antagonist darifenacin attenuates bleomycin-induced pulmonary fibrosis in rats, which may relate to the ERK/NF-κB/miRNA-21 signaling pathway.展开更多
Objective:To study the expression and significance of ECP, 25-(OH)D3 and M2 receptors in children with acute attack of asthma.Methods: Seventy children with bronchial asthma who first visited our hospital from Septemb...Objective:To study the expression and significance of ECP, 25-(OH)D3 and M2 receptors in children with acute attack of asthma.Methods: Seventy children with bronchial asthma who first visited our hospital from September 2016 to September 2018 were divided into chronic persistence group, remission group and acute attack group. Thirty healthy children who underwent physical examination in our hospital were selected and divided into control group. The levels of ECP, 25-(OH) D3 and M2 receptors were analyzed by ELISA, and Pearson correlation analysis was performed.Results: Compared with the control group, the levels of ECP and M2 receptors in chronic persistence group, remission group and acute attack group increased, while the levels of 25-(OH)D3 decreased, with statistical difference (P<0.05). The levels of ECP and M2 receptors in acute attack group were higher than those in chronic persistence group, and the levels of 25-(OH)D3 were lower than those in chronic persistence group (P<0.05). The levels of ECP and M2 receptors in acute attack group were higher than those in remission group, and the levels of 25-(OH)D3 were lower than those in remission group (P<0.05). Compared with mild children, the levels of ECP and M2 receptors increased and 25-(OH)D3 decreased in moderate and severe children (P<0.05). Compared with moderate children, the levels of ECP and M2 receptors increased and 25-(OH)D3 decreased in severe children (P<0.05).There was a negative correlation between ECP and 25-(OH)D3 (r=-0.380, P=0.038);a negative correlation between 25-(OH)D3 and M2 receptor (r=-0.448,P=0.013);and a positive correlation between ECP and M2 receptor (r=0.450,P=0.013).Conclusions:The expression of ECP and M2 receptors increased during the acute attack of bronchial asthma in children, while the expression of 25-hydroxyvitamin D3 decreased during the acute attack of bronchial asthma in children. The correlation among ECP, 25-(OH) D3 and M2 receptors is significant in the clinical diagnosis of acute attack of bronchial asthma in children.展开更多
5-HT_(1A)receptor is associated with a variety of pathophysiology of neuropsychiatric disorders.Accordingly,we have synthesized a new 5-HT_(1A) receptor ligand(HYNIC-MPP4)and labeled it with^(99m)Tc using N-(2-hydroxy...5-HT_(1A)receptor is associated with a variety of pathophysiology of neuropsychiatric disorders.Accordingly,we have synthesized a new 5-HT_(1A) receptor ligand(HYNIC-MPP4)and labeled it with^(99m)Tc using N-(2-hydroxyethyl)ethylenediaminetriacetic acid(HEDTA)as coligand.^(99m)Tc-HEDTA/HYNIC-MPP4 was prepared under pH 6 at room temperature.Biodistribution of^(99m)Tc-HEDTA/HYNIC-MPP4 in normal mice showed that this complex had moderate brain uptake(0.60%ID·g^(-1)at 2 min p.i.)and good retention.The hippocampus had the highest radioactivity uptake at 2 min p.i.(1.84%ID·g^(-1)).The ratio of Hipp/CB was 3.1 at 2 min p.i.and increased to 4.4 at 60 min p.i.After blocking with 8-hydroxy-2-(dipropylamino)tetralin,the uptake of hippocampus was decreased significantly from 1.84%ID·g^(-1) to 0.53%ID·g^(-1) at 2 min p.i.,while the cerebellum had no significant decrease.This^(99m)Tc complex could be a potent agent for 5-HT_(1A) receptor imaging.展开更多
Neuroinflammation and the NACHT,LRR,and PYD domains-containing protein 3 inflammasome play crucial roles in secondary tissue damage following an initial insult in patients with traumatic brain injury(TBI).Maraviroc,a ...Neuroinflammation and the NACHT,LRR,and PYD domains-containing protein 3 inflammasome play crucial roles in secondary tissue damage following an initial insult in patients with traumatic brain injury(TBI).Maraviroc,a C-C chemokine receptor type 5 antagonist,has been viewed as a new therapeutic strategy for many neuroinflammatory diseases.We studied the effect of maraviroc on TBI-induced neuroinflammation.A moderate-TBI mouse model was subjected to a controlled cortical impact device.Maraviroc or vehicle was injected intraperitoneally 1 hour after TBI and then once per day for 3 consecutive days.Western blot,immunohistochemistry,and TUNEL(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling)analyses were performed to evaluate the molecular mechanisms of maraviroc at 3 days post-TBI.Our results suggest that maraviroc administration reduced NACHT,LRR,and PYD domains-containing protein 3 inflammasome activation,modulated microglial polarization from M1 to M2,decreased neutrophil and macrophage infiltration,and inhibited the release of inflammatory factors after TBI.Moreover,maraviroc treatment decreased the activation of neurotoxic reactive astrocytes,which,in turn,exacerbated neuronal cell death.Additionally,we confirmed the neuroprotective effect of maraviroc using the modified neurological severity score,rotarod test,Morris water maze test,and lesion volume measurements.In summary,our findings indicate that maraviroc might be a desirable pharmacotherapeutic strategy for TBI,and C-C chemokine receptor type 5 might be a promising pharmacotherapeutic target to improve recovery after TBI.展开更多
AIM:To investigate the pathway(s)mediating rat antral circular smooth muscle contractile responses to the cholinomimetic agent,bethanechol and the subtypes of muscarinic receptors mediating the cholinergic contraction...AIM:To investigate the pathway(s)mediating rat antral circular smooth muscle contractile responses to the cholinomimetic agent,bethanechol and the subtypes of muscarinic receptors mediating the cholinergic contraction. METHODS:Circular smooth muscle strips from the antrum of Sprague-Dawley rats were mounted in muscle baths in Krebs buffer.Isometric tension was recorded.Cumulative concentration-response curves were obtained for(+)-cis- dioxolane(cD),a nonspecific muscarinic agonist,at 10^(-8)- 10^(-4)mol/L,in the presence of tetrodotoxin(TTX,10^(-7)mol/L). Results were normalized to cross sectional area.A repeat concentration-response curve was obtained after incubation of the muscle for 90 min with antagonists for M1(pirenzepine), M2(methoctramine)and M3(darifenadn)muscarinic receptor subtypes.The sensitivity to PTX was tested by the ip injection of 100 mg/kg of PTX 5 d before the experiment.The antral circular smooth muscles were removed from PTX-treated and non-treated rats as strips and dispersed smooth muscle cells to identify whether PTX-linked pathway mediated the contractility to bethanechol. RESULTS:A dose-dependent contractile response observed with bethanechol,was not affected by TTx.The pretreatment of rats with pertussis toxin decreased the contraction induced by bethanechol.Lack of calcium as well as the presence of the L-type calcium channel blocker,nifedipine,also inhibited the cholinergic contraction,with a reduction in response from 2.5±0.4 g/mm^2 to 1.2±0.4 g/mm^2(P<0.05).The dose- response curves were shifted to the right by muscarinic antagonists in the following order of affinity:darifenacin (M_3)>methocramine(M_2)>pirenzepine(M_1). CONCLUSION:The muscarinic receptors-dependent contraction of rat antral circular smooth muscles was linked to the signal transduction pathway(s)involving pertussis-toxin sensitive GTP-binding proteins and to extracellular calcium via L-type voltage gated calcium channels.The presence of the residual contractile response after the treatment with nifedipine,suggests that an additional pathway could mediate the cholinergic contraction.The involvement of more than one muscarinic receptor(functionally predominant type 3 over type 2)also suggests more than one pathway mediating the cholinergic contraction in rat antrum.展开更多
Modified constraint-induced movement therapy is an effective treatment for neurological and motor impairments in patients with stroke by increasing the use of their affected limb and limiting the contralateral limb.Ho...Modified constraint-induced movement therapy is an effective treatment for neurological and motor impairments in patients with stroke by increasing the use of their affected limb and limiting the contralateral limb.However,the molecular mechanism underlying its efficacy remains unclear.In this study,a middle cerebral artery occlusion(MCAO)rat model was produced by the suture method.Rats received modified constraint-induced movement therapy 1 hour a day for 14 consecutive days,starting from the 7^th day after middle cerebral artery occlusion.Day 1 of treatment lasted for 10 minutes at 2r/min,day 2 for 20 minutes at 2 r/min,and from day 3 onward for 20 minutes at 4 r/min.CatWalk gait analysis,adhesive removal test,and Y-maze test were used to investigate motor function,sensory function as well as cognitive function in rodent animals from the 1st day before MCAO to the 21^st day after MCAO.On the 21^st day after MCAO,the neurotransmitter receptor-related genes from both contralateral and ipsilateral hippocampi were tested by micro-array and then verified by western blot assay.The glutamate related receptor was shown by transmission electron microscopy and the glutamate content was determined by high-performance liquid chromatography.The results of behavior tests showed that modified constraint-induced movement therapy promoted motor and sensory functional recovery in the middle cerebral artery-occluded rats,but had no effect on cognitive function.The modified constraint-induced movement therapy upregulated the expression of glutamate ionotropic receptor AMPA type subunit 3(Gria3)in the hippocampus and downregulated the expression of the beta3-adrenergic receptor gene Adrb3 and arginine vasopressin receptor 1 A,Avprla in the middle cerebral artery-occluded rats.In the ipsilateral hippocampus,only Adra2 a was downregulated,and there was no significant change in Gria3.Transmission electron microscopy revealed a denser distribution the more distribution of postsynaptic glutamate receptor 2/3,which is an a-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor,within 240 nm of the postsynaptic density in the contralateral cornu ammonis 3 region.The size and distribution of the synaptic vesicles within 100 nm of the presynaptic active zone were unchanged.Western blot analysis showed that modified constraint-induced movement therapy also increased the expression of glutamate receptor 2/3 and brain-derived neurotrophic factor in the hippocampus of rats with middle cerebral artery occlusion,but had no effect on Synapsin I levels.Besides,we also found modified constraint-induced movement therapy effectively reduced glutamate content in the contralateral hippocampus.This study demonstrated that modified constraint-induced movement therapy is an effective rehabilitation therapy in middle cerebral artery-occluded rats,and suggests that these positive effects occur via the upregulation of the postsynaptic membrane a-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor expression.This study was approved by the Institutional Animal Care and Use Committee of Fudan University,China(approval No.201802173 S)on March 3,2018.展开更多
Botulinum toxin A （BTXA） has been used in several clinical trials to treat excessive glandular secretion; however, the precise mechanism of its action on the secretory function of salivary gland has not been fully e...Botulinum toxin A （BTXA） has been used in several clinical trials to treat excessive glandular secretion; however, the precise mechanism of its action on the secretory function of salivary gland has not been fully elucidated. In this study, we aimed to investigate the effect of BTXA on secretion of submandibular gland in rabbits and to identify its mechanism of action on the secretory function of salivary gland. At 12 weeks after injection with 5 units of BTXA, we found a significant decrease in the saliva flow from submandibular glands, while the salivary amylase concentration increased. Morphological analysis revealed reduction in the size of acinar cells with intracellular accumulation of secretory granules that coalesced to form a large ovoid structure. Expression of M3-muscarinic acetylcholine receptor （M3 receptor） and aquaporin-5 （AQP5） mRNA decreased after BTXA treatment, and distribution of AQP5 in the apical membrane was reduced at 1, 2 and 4 weeks after BTXA injection. Furthermore, BTXA injection was found to induce apoptosis of acini. These results indicate that BTXA decreases the fluid secretion of submandibular glands and increases the concentration of amylase in saliva. Decreased expression of M3 receptor and AQP5, inhibition of AQP5 translocation, and cell apoptosis might involve in BTXA-reduced fluid secretion of submandibular lands.展开更多
AIM： To observe the effect of Fructus Psoraleae on motility of isolated gallbladder muscle strips of guinea pigs and its mechanism. METHODS： Guinea pigs were hit to lose consciousness and the whole gallbladder was r...AIM： To observe the effect of Fructus Psoraleae on motility of isolated gallbladder muscle strips of guinea pigs and its mechanism. METHODS： Guinea pigs were hit to lose consciousness and the whole gallbladder was removed quickly. Two or three smooth muscle strips （8 mm×3mm） were cut along a longitudinal direction. The mucosa was gently removed. Every longitudinal muscle strip was suspended in a tissue chamber which was continuously perfused with 5 mL Krebs solution （37℃）, pH 7.4, and aerated with 950 mL/L 02 and 50 mL/L CO2. The isometric response was recorded with an ink-writing recorder. After 2 h equilibration under i g-load, 50 μL Fructus Psoraleae （10, 20, 70, 200, 700, 1000 g/L） was added cumulatively into the tissue chamber in turn every 2 rain to observe their effects on gallbladder muscle strips （cumulating final concentration of Fructus Psoraleae was 0.1, 0.3, 1.0, 3.0, 10.0, 20.0 g/L）. The antagonists, including 4-DAMP, benzhydramine, hexamethonium, phentolamine, verapamil and idomethine were given 2 min before Fructus Psoraleae respectively to investigate the mechanisms involved. RESULTS： Fructus Psoraleae dose-dependently increased the resting tension （r=0.992, P〈0.001）, decreased the mean contractile amplitude （r=0.970, P〈0.001） and meanwhile increased the contractile frequency of the gallbladder muscle strip in vitro （r=0.965, P〈0.001）. The exciting action of Fructus Psoraleae on the resting tension could be partially blocked by 4-DAMP （the resting tension decreased from 1.37 ± 0.41 to 0.70 ± 0.35, P〈0.001）, benzhydramine （from 1.37 ±0.41 to 0.45±0.38, P〈0.001）, hexamethonium （from 1.37 ± 0.41 to 0.94 ± 0.23, P〈0.05）, phentolamine （ from 1.37±0.41 to 0.89±0.22, P〈0.01） and verapamil （from 1.37±0.41 to 0.94±0.26, P〈0.05）. But the above antagonists had no significant effect on the action of Fructus Psoraleae-induced mean contractile amplitude （P〉0.05）. Moreover, the increase of the contractile frequency due to Fructus Psoraleae was inhibited by 4-DAMP （decreased from 8.3 ± 1.2 to 6.8 ± 0.5, P 〈 0.01） and hexamethonium （from 8.3 ±1.2 to 7.0 ± 0.9, P 〈 0.05）. Idomethine had no significant effect on the Fructus Psoraleae- induced responses （P〉 0.05）. CONCLUSION： Fructus Psoraleae enhances the motility of isolated gallbladder muscle strips from guinea pigs, in a dose-dependent manner. The effect of Fructus Psoraleae is partly related to M3, N receptor, α receptor, H1 receptor, Ca^2＋ channel, but not related to prostaglandin.展开更多
文摘Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1genes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10 -3 mol/L, 10 -4 mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10 -3 mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-κB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10 -3 mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10 -5 mol/L atropine) or NF-κB inhibitor (10 -2 mol/L PDTC) could obviously inhibit the activation of NF-κB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-1genes in pancreatic acinar cells in vitro through the activation of NF-κB.
文摘Objective: To study roles of cholinergic M2 and M3 receptors in the motility reduction of small intestine (SI) in spleen qi deficiency. Methods: 16 male SD rats were randomly divided in the control group and spleen qi deficiency group (model group)—8 rats each group;spleen qi deficiency model of the improper diet and overfatigue was established;the SI propelling rate (SIPR) was used to evaluate the SI motility;ELISA was used to measure concentrations of acetylcholine (ACh), cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in the SI tissue;immohistochemistry was employed to detect expressions of cholinergic M2 and M3 receptors. Results: Compared with those in the control group, SIPR was reduced;expression of M2 receptors was increased;and expression of M3 receptors and concentrations of cAMP and PKA were decreased, significantly, in the model group. Conclusions: Expression imbalance of cholinergic M2 and M3 receptors might contribute to the motility reduction of the SI in spleen qi deficiency.
文摘Idiopathic pulmonary fibrosis is an untreatable lethal lung disease, which is related to the aberrant proliferation of fibroblasts. M<sub>3</sub> muscarinic acetylcholine receptor (M<sub>3</sub>-mAChR) activation exerts proliferative effect on various kinds of cells. However, whether M<sub>3</sub>-mAChR inhibition has a protective effect on pulmonary fibrosis remains unexplored. A rat model of pulmonary fibrosis was established by intratracheal instillation of bleomycin. Darifenacin was used to block M<sub>3</sub>-mAChR. Histological changes were observed using Masson’s Trichrome and hematoxylin and eosin (HE) staining. Hydroxyproline was measured by Hydroxyproline detection kit. Transforming growth factor β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). In vitro, pulmonary fibroblasts were isolated from lungs of neonatal rat. After treatment, the cell viability, Hydroxyproline level was measured by MTT and Hydroxyproline detection kit respectively. The expression level of extracellular signal-regulated kinase (ERK), nuclear factor kappa-B (N-NF-κB), and microRNA-21 (miR-21) was detected by western blot or quantitative real-time PCR (qRT-PCR). Darifenacin relieved the fibrotic effects provoked by bleomycin. The expression level of hydroxyproline, TGF-β1 and TNF-α level was all downregulated after darifenacin treatment. In lung fibroblasts, darifenacin decreased cell viability and hydroxyproline level induced by bleomycin. Besides, phosphorylation-ERK and nuclear N-NF-κB protein level was downregulated, as well as miR-21 level. M<sub>3</sub>-mAChR antagonist darifenacin attenuates bleomycin-induced pulmonary fibrosis in rats, which may relate to the ERK/NF-κB/miRNA-21 signaling pathway.
基金Key projects funded by the science and technology bureau of Chenzhou city(No.CZ2013065).
文摘Objective:To study the expression and significance of ECP, 25-(OH)D3 and M2 receptors in children with acute attack of asthma.Methods: Seventy children with bronchial asthma who first visited our hospital from September 2016 to September 2018 were divided into chronic persistence group, remission group and acute attack group. Thirty healthy children who underwent physical examination in our hospital were selected and divided into control group. The levels of ECP, 25-(OH) D3 and M2 receptors were analyzed by ELISA, and Pearson correlation analysis was performed.Results: Compared with the control group, the levels of ECP and M2 receptors in chronic persistence group, remission group and acute attack group increased, while the levels of 25-(OH)D3 decreased, with statistical difference (P<0.05). The levels of ECP and M2 receptors in acute attack group were higher than those in chronic persistence group, and the levels of 25-(OH)D3 were lower than those in chronic persistence group (P<0.05). The levels of ECP and M2 receptors in acute attack group were higher than those in remission group, and the levels of 25-(OH)D3 were lower than those in remission group (P<0.05). Compared with mild children, the levels of ECP and M2 receptors increased and 25-(OH)D3 decreased in moderate and severe children (P<0.05). Compared with moderate children, the levels of ECP and M2 receptors increased and 25-(OH)D3 decreased in severe children (P<0.05).There was a negative correlation between ECP and 25-(OH)D3 (r=-0.380, P=0.038);a negative correlation between 25-(OH)D3 and M2 receptor (r=-0.448,P=0.013);and a positive correlation between ECP and M2 receptor (r=0.450,P=0.013).Conclusions:The expression of ECP and M2 receptors increased during the acute attack of bronchial asthma in children, while the expression of 25-hydroxyvitamin D3 decreased during the acute attack of bronchial asthma in children. The correlation among ECP, 25-(OH) D3 and M2 receptors is significant in the clinical diagnosis of acute attack of bronchial asthma in children.
基金Supported by the National Natural Science Foundation of China(Grant No.20401004)the Analysis and Test fund of Beijing Normal University
文摘5-HT_(1A)receptor is associated with a variety of pathophysiology of neuropsychiatric disorders.Accordingly,we have synthesized a new 5-HT_(1A) receptor ligand(HYNIC-MPP4)and labeled it with^(99m)Tc using N-(2-hydroxyethyl)ethylenediaminetriacetic acid(HEDTA)as coligand.^(99m)Tc-HEDTA/HYNIC-MPP4 was prepared under pH 6 at room temperature.Biodistribution of^(99m)Tc-HEDTA/HYNIC-MPP4 in normal mice showed that this complex had moderate brain uptake(0.60%ID·g^(-1)at 2 min p.i.)and good retention.The hippocampus had the highest radioactivity uptake at 2 min p.i.(1.84%ID·g^(-1)).The ratio of Hipp/CB was 3.1 at 2 min p.i.and increased to 4.4 at 60 min p.i.After blocking with 8-hydroxy-2-(dipropylamino)tetralin,the uptake of hippocampus was decreased significantly from 1.84%ID·g^(-1) to 0.53%ID·g^(-1) at 2 min p.i.,while the cerebellum had no significant decrease.This^(99m)Tc complex could be a potent agent for 5-HT_(1A) receptor imaging.
基金supported by grants from the National Natural Science Foundation of China, Nos. 81930031 (to JNZ), 81720108015 (to JNZ), 81901525 (to SZ), 82101440 (to DDS), 81801234 (to YZ) and 82071389 (to GLY)the Natural Science Foundation of Tianjin, Nos. 20JCQNJC01270 (to JWW), 20JCQNJC00460 (to GLY), 18JCQNJC81000 (to HTR)+4 种基金Scientific Research Project of Tianjin Education Commission (Natural Science), No. 2018KJ052 (to ZWZ)Tianjin Health and Health Committee Science and Technology Project, No. QN20015 (to JWW)the Science & Technology Development Fund of Tianjin Education Commission for Higher Education, No. 2016YD02 (to YW)Tianjin Key Science and Technology Projects of Innovative Drugs and Medical Devices, No. 19ZXYXSY00070 (to YW)the Clinical Research Fundation of Tianjin Medical University, No. 2018kylc002 (to YW)
文摘Neuroinflammation and the NACHT,LRR,and PYD domains-containing protein 3 inflammasome play crucial roles in secondary tissue damage following an initial insult in patients with traumatic brain injury(TBI).Maraviroc,a C-C chemokine receptor type 5 antagonist,has been viewed as a new therapeutic strategy for many neuroinflammatory diseases.We studied the effect of maraviroc on TBI-induced neuroinflammation.A moderate-TBI mouse model was subjected to a controlled cortical impact device.Maraviroc or vehicle was injected intraperitoneally 1 hour after TBI and then once per day for 3 consecutive days.Western blot,immunohistochemistry,and TUNEL(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling)analyses were performed to evaluate the molecular mechanisms of maraviroc at 3 days post-TBI.Our results suggest that maraviroc administration reduced NACHT,LRR,and PYD domains-containing protein 3 inflammasome activation,modulated microglial polarization from M1 to M2,decreased neutrophil and macrophage infiltration,and inhibited the release of inflammatory factors after TBI.Moreover,maraviroc treatment decreased the activation of neurotoxic reactive astrocytes,which,in turn,exacerbated neuronal cell death.Additionally,we confirmed the neuroprotective effect of maraviroc using the modified neurological severity score,rotarod test,Morris water maze test,and lesion volume measurements.In summary,our findings indicate that maraviroc might be a desirable pharmacotherapeutic strategy for TBI,and C-C chemokine receptor type 5 might be a promising pharmacotherapeutic target to improve recovery after TBI.
文摘AIM:To investigate the pathway(s)mediating rat antral circular smooth muscle contractile responses to the cholinomimetic agent,bethanechol and the subtypes of muscarinic receptors mediating the cholinergic contraction. METHODS:Circular smooth muscle strips from the antrum of Sprague-Dawley rats were mounted in muscle baths in Krebs buffer.Isometric tension was recorded.Cumulative concentration-response curves were obtained for(+)-cis- dioxolane(cD),a nonspecific muscarinic agonist,at 10^(-8)- 10^(-4)mol/L,in the presence of tetrodotoxin(TTX,10^(-7)mol/L). Results were normalized to cross sectional area.A repeat concentration-response curve was obtained after incubation of the muscle for 90 min with antagonists for M1(pirenzepine), M2(methoctramine)and M3(darifenadn)muscarinic receptor subtypes.The sensitivity to PTX was tested by the ip injection of 100 mg/kg of PTX 5 d before the experiment.The antral circular smooth muscles were removed from PTX-treated and non-treated rats as strips and dispersed smooth muscle cells to identify whether PTX-linked pathway mediated the contractility to bethanechol. RESULTS:A dose-dependent contractile response observed with bethanechol,was not affected by TTx.The pretreatment of rats with pertussis toxin decreased the contraction induced by bethanechol.Lack of calcium as well as the presence of the L-type calcium channel blocker,nifedipine,also inhibited the cholinergic contraction,with a reduction in response from 2.5±0.4 g/mm^2 to 1.2±0.4 g/mm^2(P<0.05).The dose- response curves were shifted to the right by muscarinic antagonists in the following order of affinity:darifenacin (M_3)>methocramine(M_2)>pirenzepine(M_1). CONCLUSION:The muscarinic receptors-dependent contraction of rat antral circular smooth muscles was linked to the signal transduction pathway(s)involving pertussis-toxin sensitive GTP-binding proteins and to extracellular calcium via L-type voltage gated calcium channels.The presence of the residual contractile response after the treatment with nifedipine,suggests that an additional pathway could mediate the cholinergic contraction.The involvement of more than one muscarinic receptor(functionally predominant type 3 over type 2)also suggests more than one pathway mediating the cholinergic contraction in rat antrum.
基金supported by the National Natural Science Foundation of China,No.81871841(to YLB) and No.81772453(to DSX)
文摘Modified constraint-induced movement therapy is an effective treatment for neurological and motor impairments in patients with stroke by increasing the use of their affected limb and limiting the contralateral limb.However,the molecular mechanism underlying its efficacy remains unclear.In this study,a middle cerebral artery occlusion(MCAO)rat model was produced by the suture method.Rats received modified constraint-induced movement therapy 1 hour a day for 14 consecutive days,starting from the 7^th day after middle cerebral artery occlusion.Day 1 of treatment lasted for 10 minutes at 2r/min,day 2 for 20 minutes at 2 r/min,and from day 3 onward for 20 minutes at 4 r/min.CatWalk gait analysis,adhesive removal test,and Y-maze test were used to investigate motor function,sensory function as well as cognitive function in rodent animals from the 1st day before MCAO to the 21^st day after MCAO.On the 21^st day after MCAO,the neurotransmitter receptor-related genes from both contralateral and ipsilateral hippocampi were tested by micro-array and then verified by western blot assay.The glutamate related receptor was shown by transmission electron microscopy and the glutamate content was determined by high-performance liquid chromatography.The results of behavior tests showed that modified constraint-induced movement therapy promoted motor and sensory functional recovery in the middle cerebral artery-occluded rats,but had no effect on cognitive function.The modified constraint-induced movement therapy upregulated the expression of glutamate ionotropic receptor AMPA type subunit 3(Gria3)in the hippocampus and downregulated the expression of the beta3-adrenergic receptor gene Adrb3 and arginine vasopressin receptor 1 A,Avprla in the middle cerebral artery-occluded rats.In the ipsilateral hippocampus,only Adra2 a was downregulated,and there was no significant change in Gria3.Transmission electron microscopy revealed a denser distribution the more distribution of postsynaptic glutamate receptor 2/3,which is an a-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor,within 240 nm of the postsynaptic density in the contralateral cornu ammonis 3 region.The size and distribution of the synaptic vesicles within 100 nm of the presynaptic active zone were unchanged.Western blot analysis showed that modified constraint-induced movement therapy also increased the expression of glutamate receptor 2/3 and brain-derived neurotrophic factor in the hippocampus of rats with middle cerebral artery occlusion,but had no effect on Synapsin I levels.Besides,we also found modified constraint-induced movement therapy effectively reduced glutamate content in the contralateral hippocampus.This study demonstrated that modified constraint-induced movement therapy is an effective rehabilitation therapy in middle cerebral artery-occluded rats,and suggests that these positive effects occur via the upregulation of the postsynaptic membrane a-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor expression.This study was approved by the Institutional Animal Care and Use Committee of Fudan University,China(approval No.201802173 S)on March 3,2018.
基金grants from the National Natural Science Foundation of China （Nos. 30973336 and 81100762）.
文摘Botulinum toxin A （BTXA） has been used in several clinical trials to treat excessive glandular secretion; however, the precise mechanism of its action on the secretory function of salivary gland has not been fully elucidated. In this study, we aimed to investigate the effect of BTXA on secretion of submandibular gland in rabbits and to identify its mechanism of action on the secretory function of salivary gland. At 12 weeks after injection with 5 units of BTXA, we found a significant decrease in the saliva flow from submandibular glands, while the salivary amylase concentration increased. Morphological analysis revealed reduction in the size of acinar cells with intracellular accumulation of secretory granules that coalesced to form a large ovoid structure. Expression of M3-muscarinic acetylcholine receptor （M3 receptor） and aquaporin-5 （AQP5） mRNA decreased after BTXA treatment, and distribution of AQP5 in the apical membrane was reduced at 1, 2 and 4 weeks after BTXA injection. Furthermore, BTXA injection was found to induce apoptosis of acini. These results indicate that BTXA decreases the fluid secretion of submandibular glands and increases the concentration of amylase in saliva. Decreased expression of M3 receptor and AQP5, inhibition of AQP5 translocation, and cell apoptosis might involve in BTXA-reduced fluid secretion of submandibular lands.
基金Supported by Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province and the Natural Science Foundation of Gansu Province, No.ZS021-A25-059-Y
文摘AIM： To observe the effect of Fructus Psoraleae on motility of isolated gallbladder muscle strips of guinea pigs and its mechanism. METHODS： Guinea pigs were hit to lose consciousness and the whole gallbladder was removed quickly. Two or three smooth muscle strips （8 mm×3mm） were cut along a longitudinal direction. The mucosa was gently removed. Every longitudinal muscle strip was suspended in a tissue chamber which was continuously perfused with 5 mL Krebs solution （37℃）, pH 7.4, and aerated with 950 mL/L 02 and 50 mL/L CO2. The isometric response was recorded with an ink-writing recorder. After 2 h equilibration under i g-load, 50 μL Fructus Psoraleae （10, 20, 70, 200, 700, 1000 g/L） was added cumulatively into the tissue chamber in turn every 2 rain to observe their effects on gallbladder muscle strips （cumulating final concentration of Fructus Psoraleae was 0.1, 0.3, 1.0, 3.0, 10.0, 20.0 g/L）. The antagonists, including 4-DAMP, benzhydramine, hexamethonium, phentolamine, verapamil and idomethine were given 2 min before Fructus Psoraleae respectively to investigate the mechanisms involved. RESULTS： Fructus Psoraleae dose-dependently increased the resting tension （r=0.992, P〈0.001）, decreased the mean contractile amplitude （r=0.970, P〈0.001） and meanwhile increased the contractile frequency of the gallbladder muscle strip in vitro （r=0.965, P〈0.001）. The exciting action of Fructus Psoraleae on the resting tension could be partially blocked by 4-DAMP （the resting tension decreased from 1.37 ± 0.41 to 0.70 ± 0.35, P〈0.001）, benzhydramine （from 1.37 ±0.41 to 0.45±0.38, P〈0.001）, hexamethonium （from 1.37 ± 0.41 to 0.94 ± 0.23, P〈0.05）, phentolamine （ from 1.37±0.41 to 0.89±0.22, P〈0.01） and verapamil （from 1.37±0.41 to 0.94±0.26, P〈0.05）. But the above antagonists had no significant effect on the action of Fructus Psoraleae-induced mean contractile amplitude （P〉0.05）. Moreover, the increase of the contractile frequency due to Fructus Psoraleae was inhibited by 4-DAMP （decreased from 8.3 ± 1.2 to 6.8 ± 0.5, P 〈 0.01） and hexamethonium （from 8.3 ±1.2 to 7.0 ± 0.9, P 〈 0.05）. Idomethine had no significant effect on the Fructus Psoraleae- induced responses （P〉 0.05）. CONCLUSION： Fructus Psoraleae enhances the motility of isolated gallbladder muscle strips from guinea pigs, in a dose-dependent manner. The effect of Fructus Psoraleae is partly related to M3, N receptor, α receptor, H1 receptor, Ca^2＋ channel, but not related to prostaglandin.
