摘要
目的探讨益气活血通脉颗粒治疗调控小胶质细胞表达MMP-9与TIMP1蛋白介导脑缺血再灌注损伤的作用机制。方法采用BV2小胶质细胞进行氧糖剥夺和复氧实验,设置模型组、空白组、MMP-9 siRNA组、益气活血通脉颗粒组、MMP-9 siRNA+益气活血通脉颗粒组。除空白组外,其余4组均置于氧糖剥夺罐进行氧糖剥夺/复氧实验。CCK8检测各组细胞增殖能力。Western blot检测各组小胶质细胞中MMP-9与TIMP1蛋白表达水平。结果与空白组比较,模型组小胶质细胞存活率明显降低(P<0.01),与模型组比较,MMP-9 siRNA组、益气活血通脉颗粒组与MMP-9 siRNA+益气活血通脉颗粒组小胶质细胞存活率升高(P<0.05或P<0.01)。与空白组比较,模型组小胶质细胞MMP-9蛋白表达水平升高(P<0.01),TIMP1蛋白表达水平降低(P<0.01),MMP-9 siRNA组、益气活血通脉颗粒组与MMP-9 siRNA+益气活血通脉颗粒组MMP-9蛋白表达水平降低(P<0.05或P<0.01),TIMP1蛋白表达水平升高(P<0.05或P<0.01)。结论益气活血通脉颗粒能够调节MMP-9/TIMP1平衡,有助于抵抗缺氧复氧导致的小胶质细胞损伤。
Objective To investigate the mechanism of Yiqi Huoxue Tongmai granule in regulating microglia to mediate cerebral ischemia-reperfusion injury.Methods The BV2 microglia(BV2)model of oxygen and sugar deprivation/reoxygenation was adopted.The experiment objects were divided into five groups for experimental research,including the model group,the blank group,the MMP-9 siRNA group,the Yiqi Huoxue Tongmai granules drug-containing serum intervention group and the MMP-9 siRNA+Yiqi Huoxue Tongmai granules drug-containing serum intervention group.Except blank group,the other four groups were placed in oxygen and sugar deprivation tank for oxygen and sugar deprivation/reoxygenation experiment.Cell proliferation was detected by Cell Counting Kit-8.The expression levels of MMP-9 and TIMP1 in microglia were detected by Western blot.Results The survival rate of microglia cells in the model group was significantly reduced(P<0.01)in comparison with the blank group,and the survival rate of microglia cells in MMP-9 siRNA group,Yiqi Huoxue Tongmai granules drug-containing serum intervention group and MMP-9 siRNA+Yiqi Huoxue Tongmai granules drug-containing serum intervention group was significantly higher(P<0.05 or P<0.01)in comparison with the model group.Compared with blank group,MMP-9 protein expression level of microglia cells in model group was significantly increased(P<0.01),while TIMP1 protein expression level was significantly decreased(P<0.01).The protein expression level of MMP-9 in MMP-9 siRNA group,Yiqi Huoxue Tongmai granules drug-containing serum intervention group and MMP-9 siRNA+Yiqi Huoxue Tongmai granules drug-containing serum intervention group was significantly decreased(P<0.05 or P<0.01),while the protein expression level of TIMP1 was significantly increased(P<0.05 or P<0.01).Conclusion Yiqi Huoxue Tongmai granules can regulate the imbalance of MMP-9/TIMP1 and help to resist microglia cell damage caused by hypoxia and reoxygenation.
出处
《浙江临床医学》
2024年第6期795-798,共4页
Zhejiang Clinical Medical Journal
基金
浙江省中医药科技计划项目(2021ZB101)
浙江省卫生健康科技计划青年创新人才项目(2022RC214)。