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Objective To study the expression of vesicular GABA transporter and vesicular glutamate transporter 1 in de‐pression .Methods Mice was divided into control group and defeat group stochastically .By social defeat model and social avoidance , the defeat group was divided into two groups:susceptible group and unsusceptible group .Synaptic proteins were extracted respec‐tively from the 3 groups .We detected the expression abundance of VGAT and VGLUT1 by Western blot .Results Compared with the control group ,in susceptible group ,the residence time in the contact area was significantly reduced ,and the residence time in the corner area was significantly increased ,with statistical difference(P0 .05) .In the striatum ,although the expression levels of VGAT and VGLUT1 were increased in susceptible group ,but in unsusceptible group ,the expression of these proteins also increased significantly .Conclusion The prefrontal cortex and hippo‐campus excitability and inhibitory vesicle transport were changed in depression ,which may relate to the transcription disorder .
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Objective To investigate the effect of minocycline on the cognition and expressions of brain-derived neurotrophic factor (BDNF), apoptosis related factor Bcl-2 and Bax in hippocampus of rats with Alzheimer’s disease (AD). Methods The rat model was established by microinjection of Aβ25-35 into lateral ventricle. Thirty healthy male SD rats were randomly divided into three groups:control group, model group and minocycline treatment group. Normal saline 1 mL/(kg·d) was intraperitoneally injected in control group and model group. The minocycline treatment group was intraperitoneally injected with minocycline 50 mg/(kg · d) for 14 days. Morris water maze was used to detect the behaviors of animals. The expressions of BDNF, Bcl-2 and Bax in hippocampus were measured by Western blotting and enzyme linked immunosorbent assay (ELISA). The apoptosis of neurons was detected by TdT-mediated dUTP nick-end labeling (TUNEL). Results Minocycline greatly improved the behaviors of AD rats, up-regulated the expressions of BDNF and Bcl-2, and down-regulated the expression of Bax in hippocampus, and reduced cell apoptosis. Conclusion Minocycline plays a protective role in neural function by promoting the growth of neurons and inhibiting the neuronal apoptosis.
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OBJECTIVE To explore the effect of chloride channels on the neuronal injury following cerebral ischemia. METHODS Tweleve day in vitro (12dIV) neurons in rats were randomly divided into normal control, 3-morpholinosydnonimine (SIN-1, 1.0 mmol·L~(-1) for 18 h) group, SIN-1+4-acetamido-4'-isothiocyanatostilbene-2,2'-disulphonic acid(SITS, 0.5 mmol·L~(-1)) group and SIN-1+4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS, 0.1 mmol·L~(-1)) group. Drugs were added with SIN-1 simultaneously and coincubated for 18 h. The neuronal apoptosis and morphological changes were detected with Hoechst 33258. Chloride channels(ClC)-2/ClC-3 were analyzed with immunofluorescence, the chloride channel currents were recorded with whole cell patch-clamp technique. RESULTS Hoechst 33258 staining showed that the apoptotic percentages were (18.61±0.59) %, (50.43±0.56)%, (23.37±0.52)% and (23.37±0.84)% in normal control group, SIN-1 group, SIN-1+SITS or SIN-1+DIDS groups, respectively. ClC-2/ClC-3 were positively expressed in normal neurons. The currents in neurons exposed to SIN-1 were increased about 55%-56%, SITS and DIDS, two kinds of chloride channel blockers could inhibited the currents about 50%-60% and 30%-40%, respectively. CONCLUSION Voltage-dependent chloride channel maybe participate in the neuronal apoptosis induced by NO, and the activities of chloride channels are perhaps involved in the cerebral ischemic injury.
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Aim To observe the protective effects of SITS and DIDS,two kinds of chloride channel blockers,on hippocampal neuronal damage induced by NO in culture.Methods The cultures were divided into three groups:control group,NO treatment group,NO treatment plus chloride channel blocker group. The cultures were detected with the methods of morphological stain (Hoechst 33258),and the apoptotic neurons and neuronal viabilities were observed through MTT quantitative analysis. The activated caspase-3 was analyzed with western blot.Results There were significant protective effects of SITS and DIDS on neuronal damage with dose-dependence.Conclusions Chloride channel blockers have some protective effects against neuronal injury induced by NO.
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Aim To observe the action of estrone on the hippocampal neuronal apoptosis induced by NMDA in vitro and to analyse the mechanism underlying the neuroprotection of estrone. Methods The methods of both morphological analysis and cell viabilities were used, and Western blot was also used to analyce the role of antitoxiciy of estrone. Results The cultures were conformed with fluorescent dye of Hoechst 33258 and NeuN. About 31.6% neurons were induced into apoptosis by NMDA(300 ?mol?L~ -1 +glycine 5 ?mol?L~ -1 ,compared with control group, P0.05). MTT test also showed that estrone can protect cultures from apoptosis induced by NMDA, it can raise the cell viabilities exposed to NMDA. Western blot showed that activities of caspase-3 can be inhibited by estrone, the segments of caspase-3 can diminished after application of estrone together with NMDA. Conclusion Estrone had neuronal protection, this role maybe relate to the inhibition of caspase-3, a common pathway in apoptosis. The results can provide some clues to the explovation of the clinical use of estrone and to the development of a neuronal protective drug.
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Objective To study the effect of S2-protein from SARS coronavirus on the chloride channel currents in A549 cells and its possible cellular mechanisms. Methods The chloride channel currents were recorded in cultured A549 cells by using the whole-cell mode of patch clamp techniques. The experiments were divided into four groups: Control group: chloride channel currents were recorded in untreated A549 cells; S2 protein group: currents were recorded in A549 cells treated with S2 protein (final concentration 50?g/ml); calphostin C + S2 protein group: the effect of S2 protein on the currents in A549 cells pretreated with calphostin C (0.1mmol/L) for 10 minutes; SB203580+S2 protein group: the effect of S2 protein on the currents was examined with the solution containing SB203580 (20?mol/L). Results The currents of chloride channel in normal A549 cells showed outwardly rectifying properties and were insensitive to both TEA and amiloride, but were significantly inhibited both by SITS and DIDS (P
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Objective Effects of aqueous extract from Lishi No.5 formula on concentration of intracellular free-calcium fluorescent intensity in neurons. Methods Hippocampus neurons of 1-day newborn SD rats were cultured with conventional culture technique. The cells cultured for 9-12 days were used for experiment. Intracellular free calcium fluorescent intensity of neurons cultured under different conditions was assayed with confocal microscopic calcium image technique after loading of Fluo-3/AM. Results Free-calcium concentration was enhanced by aqueous extract from Lishi No.5 formula, this concentration was up to 126.35?9.35nmol/L, but the concentration is at normal scope; L-type calcium channel blocker Nifedipine may block the effect of aqueous extract from Lishi No.5 formula enhancing intracellular free-calcium concentration in part, it make intracellular free-calcium concentration down to 90.75?10.15nmol/L, but Nifedipine itself also decreased markedly free-calcium concentration to 40.65?5.65nmol/L. NMDA increase Markedly calcium fluorescent intensity, the effects of NMDA was decreased notably after pre-treating with aqueous extract from Lishi No.5 formula. The effect of MK-801, an antagonist of NMDA receptor, on inhibition of NMDA increasing free calcium fluorescent intensity was significantly reduced after pretreatment of aqueous extract from Lishi No.5 formula. Conclusion Aqueous extract from Lishi No.5 formula maybe bidirectional adjust intracellular free calcium concentration via enhancing L-type calcium channel activity and blocking NMDA receptor in part.