脑缺血再灌注后大鼠脑组织中自噬-炎症串扰相关蛋白表达变化

Expression changes of autophagy-inflammation crosstalk related proteins in brain tissues after cerebral ischemia-reperfusion

目的观察脑缺血再灌注后大鼠脑组织中自噬-炎症串扰相关蛋白的表达变化,探讨脑梗死的发病机制。方法将15只SD雄性大鼠随机分为模型组8只与假手术组7只,模型组参照Longa线栓法制备缺血再灌注模型,假手术组不阻塞大脑中动脉;经Longa评分、伊文思蓝染色检测模型成功后,采用Western blotting法检测脑组织中的自噬相关蛋白[自噬相关蛋白16-1(ATG16L1)、Beclin-1、LC-3β]和炎症相关蛋白[Toll样受体4(TLR4)、NLRP3、IL-1]。结果模型组脑组织中ATG16L1蛋白相对表达量增高,而LC-3β、Beclin-1蛋白相对表达量降低(P均<0.05);与假手术组比较,模型组脑组织中TLR4、NLRP3和IL-1蛋白相对表达量升高(P均<0.05)。结论脑缺血再灌注损伤后大鼠脑组织中自噬-炎症串扰反应异常,即自噬相关蛋白被抑制而炎症相关蛋白被激活;这可能是脑梗死脑损伤加重、神经功能恢复受阻的原因。

Objective To observe the expression changes of autophagy-inflammation crosstalk related proteins in the brain tissues of rats after cerebral ischemia-reperfusion injury and to explore the pathogenesis of cerebral infarction.Methods Fifteen male SD rats were randomly divided into the model group(n=8)and sham operation group(n=7).The ischemia-reperfusion rat model was made by Longa thread bolt method in the model group,and the middle cerebral artery of rats in the sham operation group was not blocked.After the model was proved to be successful by Longa score and Evans blue staining,Western blotting was used to detect the expression of autophagy-related proteins[autophagy-related 16 like 1(Atg16L1),Beclin-1,light chain 3β(LC3β)]and inflammation-related proteins[Toll-like receptor 4(TLR4),NLRP3,and interleukin 1β(IL-1β)]in the brain tissues.Results In the model group,the relative expression of Atg16L1 protein increased,while the relative expression levels of LC3βand Beclin-1 protein decreased(all P<0.05);compared with the sham operation group,the relative expression levels of TLR4,NLRP3 and IL-1βprotein increased in the model group,with statistically significant difference(all P<0.05).Conclusion The autophagy-inflammation crosstalk in the brain of rats after cerebral ischemia-reperfusion injury is abnormal,and the autophagy-related proteins are inhibited and inflammation-related proteins are activated,which might be the reason for the aggravation of cerebral infarction and the obstruction of neurological function recovery.