Gap19介导Cx43半通道抑制JAK2/STAT3通路活化对急性脑梗死大鼠的保护作用

Protective effects of Gap19-mediated Cx43 hemichannel inhibition and activation of JAK2/STAT3 pathway on rats with acute cerebral infarction

目的探究Gap19通过介导Cx43半通道活性和JAK2/STAT3通路对急性脑梗死(ACI)大鼠的影响。方法24只SD大鼠随机分为3组:对照组、MCAO组和MCAO+Gap19组,每组8只。建立ACI大鼠模型(MCAO)和氧葡萄糖剥夺(OGD)体外模型。TTC染色检测大鼠脑梗死面积;mNSS法检测大鼠神经功能缺损;CCK-8检测星形胶质细胞活力;Western blot检测NVU模型Cx43、Cleaved caspase-3、Bcl、Bax和JAK2/STAT3通路相关基因的蛋白表达;溴化乙锭摄取实验检测星形胶质细胞中半通道活性;免疫荧光法检测星形胶质细胞中pSTAT3和Cx43的共定位。结果MCAO处理导致大鼠脑梗死面积和神经功能缺损均显著上调(P<0.05),Gap19处理显著下调大鼠脑梗死面积和神经功能缺损(P<0.05)。OGD处理导致NVU模型Cx43的蛋白表达和半通道活性显著上调(P<0.05),细胞活力显著下调(P<0.05),促进细胞凋亡;Gap19处理显著上调OGD处理后NVU模型细胞活力(P<0.05),显著下调Cx43的蛋白表达和星形胶质细胞的半通道活性(P<0.05),抑制细胞凋亡,促进JAK2/STAT3通路活化,其中Cx43与pSTAT3存在共定位关系。结论Gap19通过抑制Cx43半通道和活化JAK2/STAT3通路发挥对ACI大鼠的保护作用。

Objective To explore the effect of Gap19 on acute cerebral infarction(ACI)rats by mediating the activity of Cx43 hemichannel and JAK2/STAT3 pathway.Methods Twenty-four Sprague-Dawley(SD)rats were randomly divided into three groups with 8 rats in each group:control group,MCAO group and MCAO+Gap19 group.MCAO rat models and and oxygen glucose deprivation(OGD)in vitro models were established.2,3,5,tripheyl tetrazolium chloride(TTC)staining was used to detect cerebral infarct area in rats;mNSS method was utilized to detect neurological deficits in rats;Astrocyte viability was measured by CCK-8 assay;Western blot was used to detect the protein expression of Cx43,cleaved caspase-3,Bcl,Bax and JAK2/STAT3 pathway related molecules in NVU model;Ethidium bromide uptake assay was performed to detect hemichannel activity in astrocytes;Co-localization of pSTAT3 and Cx43 in astrocytes was detected by immunofluorescence.Results Rats in MCAO group displayed significantly increased cerebral infarct area and neurological deficits(P<0.05),and Gap19 treatment significantly reduced the cerebral infarct area and neurological deficits in rats(P<0.05).OGD treatment resulted in significant up-regulation and hemichannel activity of Cx43 in NVU model(P<0.05),significantly reduced cell viability(P<0.05),and promoted apoptosis;Gap19 treatment significantly increased the cell viability of the NVU model cells after OGD treatment(P<0.05),significantly reduced the protein level of Cx43 and the hemichannel activity of astrocytes(P<0.05),inhibited apoptosis and promoted activation of JAK2/STAT3 pathway.In addition,Cx43 and pSTAT3 have a co-localization relationship.Conclusion Gap19 exerts a protective effect on rats with ACI by inhibiting the Cx43 hemichannel and activating the JAK2/STAT3 pathway.