刺槐素通过自噬调控ROS/NLRP3信号通路对脑缺血再灌注损伤发挥保护作用  

Acacetin protects against cerebral ischemia-reperfusion injury by regulating ROS/NLRP3 signaling pathway via autophagy

作　　者：补娟[1] 纪国庆[2] 叶勒丹·马汉 王兆霞 吴选霞 牛晓珊[2] BU Juan;JI Guoqing;YELEDAN Mahan(Center for Medical Research and Transformation,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院医学研究与转化中心,新疆乌鲁木齐830001 [2]新疆维吾尔自治区人民医院全科医疗,新疆乌鲁木齐830001

出　　处：《中风与神经疾病杂志》2023年第2期99-102,共4页Journal of Apoplexy and Nervous Diseases

基　　金：新疆维吾尔自治区自然科学基金(No.2020D01C089);新疆维吾尔自治区人民医院院内项目(No.20190307)。

摘　　要：目的基于自噬/ROS/NLRP3炎症小体信号通路,探讨刺槐素(Acacetin)对氧糖剥夺再灌注(OGD/R)损伤后的小胶质细胞保护作用机制。方法培养小鼠小胶质细胞(BV2),分为正常对照组、OGD模型组和OGD+Acacetin组(10μmol)。缺氧6 h复氧24 h后采用4-甲基偶氮唑蓝(MTT)检测BV2细胞活性;乳酸脱氢酶(LDH)法检测细胞的死亡率;活性氧(ROS)试剂盒测定细胞内ROS水平;Western blot检测LC3-Ⅱ、LC3-Ⅱ/LC3-Ⅰ、beclin-1、NLRP3、caspase-1和IL-1β等蛋白的表达。结果刺槐素能增加OGD/R损伤后小胶质细胞的存活率,减少LDH的释放,降低ROS的生成,增加LC3-Ⅱ和beclin-1蛋白的表达,进一步下调NLRP3、caspase-1和IL-1β的表达。结论Acacetin能减轻OGD/R后小胶质细胞的损伤从而对脑缺血再灌注损伤发挥保护作用,其作用机制可能与抑制ROS的产生、激活自噬、抑制NLRP3炎症小体有关。Objective To explore the protective effects of Acacetin on mouse microglial cells injured by oxygen-glucose deprivation/reperfusion(OGD/R)based on autophagy/ROS/NLRP3 inflammasomes signaling pathway.Methods Mouse microglial cells(BV2)were divided into normal control group,OGD model group and OGD+Acacetin group(10μmol).After hypoxia for 6 hours and reoxygenation for 24 hours,the viability of BV2 cells was detected by 4-methylazolazolium blue(MTT)method.The death rate of cell was measured by lactate dehydrogenase(LDH)method.Reactive oxygen species(ROS)assay kit was used to measure the intracellular ROS level.The expression of LC3-Ⅱ,LC3-Ⅱ/LC3-Ⅰ,beclin-1,NLRP3,caspase-1 and IL-1βwere analyzed by western blot.Results Acacetin increased the survival rate of microglia cells,and reduced the release of LDH and production of ROS.Acacetin also increased the expression of LC3-Ⅱ,and further to decrease the expression of NLRP3,caspase-1 and IL-1β.Conclusions Acacetin protected against cerebral ischemia-reperfusion injury by attenuating the injury of microglia after OGD/R.Its protective mechanism may be related to inhibiting the production of ROS,activating autophagy,and inhibiting the NLRP3 inflammasome.

关 键 词：刺槐素 氧糖剥夺再灌注 小胶质细胞 NLRP3炎症小体 自噬 

分 类 号：R743.3[医药卫生—神经病学与精神病学]