1-甲基色氨酸对大鼠脑缺血再灌注模型小胶质细胞活化的干预作用

Effect of 1-Methyltryptophan on the Activation of Microglia in Rat Model of Cerebral Ischemia Reperfusion

目的研究IDO抑制剂1-甲基色氨酸(1-methyltryptophan,1-MT)对大鼠脑缺血再灌注模型小胶质细胞活化的干预作用及相关炎症因子表达的影响。方法 SD大鼠分为假手术组、模型组和1-MT组。使用大脑中动脉闭塞法(middle cerebral artery occlusion,MCAO)构建缺血再灌注模型。对大鼠进行神经功能缺损评分,TTC法测定脑梗死体积,HE染色检测梗死区域的病理形态变化,免疫组化法检测梗死区域IBA-1和GFAP的表达,免疫荧光技术检测NeuN阳性细胞数,Western blot法检测组织中IL-1β、TNF-α、NF-κB的表达水平。体外培养BV2细胞,采用氧糖剥夺法(oxygen glucose deprivation,OGD)模拟缺血缺氧条件,设置对照组、模型组和1-MT组,Western blot法检测小胶质细胞标志物IBA-1、IL-1β、NF-κB以及TNF-α的表达水平。结果 1-MT可以显著改善MCAO大鼠的神经功能,缩小梗死面积,抑制小胶质细胞的活化和相关炎症因子的表达。细胞实验中,1-MT可以显著下调OGD-BV2细胞中IBA-1的表达水平和IL-1β、NF-κB、TNF-α的水平。结论 IDO抑制剂1-MT对于大鼠脑缺血再灌注模型有着很好的保护作用,其作用机制与小胶质细胞的活化抑制以及相关炎症因子的表达有关。

OBJECTIVE To study the IDO inhibitor of 1-methyltryptophan (1-MT) intervented on microglia activation and inflammatory factor expression on cerebral ischemia reperfusion rat.METHODS SD rats were divided into sham operation group,model group,and 1-MT group.The model of ischemia reperfusion was constructed by the middle cerebral artery occlusion (MCAO) method.The nerve function defect was scored in rats.Determination of cerebral infarction volume by TTC method.The pathological changes of infarct area were detected by HE staining.Immunohistochemical method was used to detect the expression of IBA-1 and GFAP in the infarct area.The number of NeuN positive cells was detected by immunofluorescence.The expression of IL-1β,NF-κB,TNF-α in the tissue was detected by Western blot.BV2 cells were cultured in vitro,and oxygen glucose deprivation(OGD) was used to simulate the conditions of ischemic and anoxia.The control group,model group and 1-MT group were set up.The expression levels of IBA-1,IL-1β,NF-κB,TNF-α were detected by Western blot.RESULTS 1-MT could significantly improve the neural function of MCAO rats,reduce the infarct size,inhibit the activation of microglia and the expression of related inflammatory factors.1-MT could significantly reduce the IBA-1 expression of IBA-1,IL-1β,NF-κB and TNF-α in OGD-BV2 cells.CONCLUSION IDO inhibitor 1-MT has a good protective effect on the cerebral ischemia-reperfusion in rats,and its mechanism is related to the inhibition of microglia and the expression of related inflammatory factors.