溶血磷脂酸调控RhoA/ROCK2信号通路对大鼠脑缺血再灌注后的影响

Effect of lysophosphatidic acid-regulated RhoA/ROCK2 signaling pathway on cerebral ischemia and reperfusion in rats

目的分析溶血磷脂酸(LPA)调控RhoA/ROCK2信号通路对脑缺血再灌注后的影响。方法使用线栓法成功构建20只脑动脉栓塞大鼠,缺血2 h后再行灌注,灌注24 h后完成脑缺血再灌注模型,将其按照随机数字表法分为两组,每组各10只,分别为模型组和LPA组。LPA组每日腹腔注射50μmol/L的LPA,模型组每日腹腔注射等量0.9%氯化钠溶液,同时随机纳入10只健康大鼠作为对照组,各组均标准饲养14 d。比较各组大鼠脑神经细胞活力、迁移、凋亡情况,并比较各组大鼠14 d后脑神经细胞炎症因子[白细胞介素(IL)-6、IL-1β、肿瘤坏死因子α(TNF-α)]水平、细胞内RhoA、ROCK2蛋白和mRNA表达情况。结果标准饲养7 d和14 d后,模型组大鼠的脑神经细胞活力和迁移个数均低于对照组,LPA组大鼠的脑神经细胞活力和迁移个数均高于模型组,差异均有统计学意义(P<0.05);且标准饲养14 d后,LPA组大鼠的脑神经细胞活力和迁移个数高于7 d时,而模型组大鼠14 d的脑神经细胞活力和迁移个数低于7 d时,差异均有统计学意义(P<0.05)。标准饲养7 d和14 d后,模型组大鼠的脑神经细胞凋亡率均高于对照组,LPA组大鼠的脑神经细胞凋亡率低于模型组,差异均有统计学意义(P<0.05);标准饲养14 d后,LPA组大鼠的脑神经细胞凋亡率低于7 d,而模型组大鼠14 d的脑神经细胞凋亡率高于7 d时,差异均有统计学意义(P<0.05)。标准饲养14 d后,模型组大鼠的脑神经细胞IL-6、IL-1β、TNF-α水平均高于对照组,LPA组大鼠的脑神经细胞IL-6、IL-1β、TNF-α水平低于模型组,差异均有统计学意义(P<0.05)。标准饲养14 d后,模型组大鼠的脑神经细胞内RhoA、ROCK2蛋白和mRNA均低于对照组,LPA组大鼠的脑神经细胞内RhoA、ROCK2蛋白和mRNA均高于模型组,差异均有统计学意义(P<0.05)。结论LPA可通过调控RhoA/ROCK2信号通路提高脑神经细胞活性,促进迁移,抑制凋亡,缓解炎症反应。

Objective To analyze the effect of lysophosphatidic acid(LPA)regulating RhoA/ROCK2 signaling pathway on cerebral ischemia and reperfusion.Methods Twenty rats with cerebral artery embolization were successfully constructed using the thread embolization method.The rats were reperfused after 2 hours of ischemia,and the cerebral ischemia reperfusion model was completed after 24 hours of perfusion.They were divided into 2 groups according to the random number table method,each with 10 rats,the model group and the LPA group.The LPA group was intraperitoneally injected with 50μmol/L LPA daily,and the model group was intraperitoneally injected with the same amount of normal saline daily.At the same time,10 healthy rats were randomly included as the control group.Each group was fed for 14 days.The viability,migration,and apoptosis of brain nerve cells in each group were compared,and the levels of inflammatory factors[interleukin(IL)-6,IL-1βand tumor necrosis factorα(TNF-α)]in brain nerve cells,the expression of RhoA and ROCK2 protein and mRNA in the cells were compared after 14 days.Results The brain nerve cell activity and migration number of rats in the model group were lower than the control group after standard feeding for 7 days and 14 days,and the LPA group were higher than the model group,the differences were statistically significant(P<0.05);after 14 days of standard feeding,the viability and migration number of cerebral nerve cells in LPA group were higher than those at 7 days,while the viability and migration number of brain nerve cells in model group rats at 14 days were lower than 7 days,the differences were statistically significant(P<0.05).After standard feeding for 7 days and 14 days,the apoptosis rate of brain nerve cells in the model group was higher than that of the control group,and the LPA group was lower than the model group,the differences were statistically significant(P<0.05);after 14 days of standard feeding,the apoptosis rate of brain nerve cells in LPA group was lower than that at 7 days,while the apoptosis rate of brain nerve cells in model group rats at 14 days was higher than that at 7 days,the differences were statistically significant(P<0.05).After standard feeding for 14 days,the levels of IL-6,IL-1βand TNF-αin the brain nerve cells of the model group were higher than those of the control group,the LPA group were lower than those of the model group,the differences were statistically significant(P<0.05).After standard feeding for 14 days,the expression of RhoA,ROCK2 protein and mRNA in the brain nerve cells of the model group were lower than those in the control group,and the RhoA,ROCK2 protein and mRNA in the brain nerve cells of the LPA group were higher than those in the model group,and the differences were statistically significant(P<0.05).Conclusion LPA can increase brain nerve cell activity by regulating RhoA/ROCK2 signaling pathway,promote migration,inhibit apoptosis,and alleviate inflammatory response.