黄芪甲苷对小胶质细胞NLRP3/Caspase-1通路相关蛋白表达的影响  

Effects of Astragaloside IV on NLRP3/Caspase-1 Pyroptosis Pathway in Microglia

作　　者：杨治伦 陈小江[1] 陈飞飞 原茜倩 马全瑞[1] 秦毅[1] YANG Zhilun;CHEN Xiaojiang;CHEN Feifei;YUAN Qianqian;MA Quanrui;QIN Yi(Department of Human Anatomy and Histo-Embryology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院人体解剖与组织胚胎学系,银川750004

出　　处：《宁夏医科大学学报》2023年第5期451-455,共5页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金项目(NZ2019AAC03089)。

摘　　要：目的 探讨黄芪甲苷(astragaloside Ⅳ,AS-Ⅳ)对小鼠小胶质细胞焦亡通路相关蛋白的影响及机制。方法 体外培养小鼠小胶质细胞(BV2),设置Control组(正常对照组),以20 U·mL^(-1)凝血酶激活小胶质细胞作为凝血酶组,20 U·mL^(-1)凝血酶+不同浓度的AS-Ⅳ(1、5和10μmol·L^(-1))作为实验组。采用免疫印迹及免疫荧光法检测BV2细胞中焦亡相关因子NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、含半胱氨酸的天冬氨酸蛋白酶水解1(Caspase-1)、胞膜穿孔蛋白D-N端(GSDMD-N)表达量变化,ELISA法检测细胞培养液中焦亡下游分泌物[白细胞介素(IL-18、IL-1β)]的含量。结果 免疫印迹及免疫荧光结果显示,与Control组比较,凝血酶组BV2细胞内焦亡相关因子(ASC、NLRP3、Caspase-1、GSDMD-N)蛋白表达量均升高(P均<0.05);与凝血酶组比较,不同浓度AS-Ⅳ实验组BV2细胞内焦亡相关因子(ASC、NLRP3、Caspase-1、GSDMD-N)表达量均降低(P均<0.05),并且5、10μmol·L^(-1)AS-Ⅳ组较1μmol·L^(-1)AS-Ⅳ组均降低(P均<0.05)。ELISA结果显示,凝血酶组IL-18与IL-1β含量相较于Control组增加(P均<0.05),而不同浓度AS-Ⅳ实验组IL-18与IL-1β含量相较于凝血酶组均减少(P均<0.05),并且5、10μmol·L^(-1)AS-Ⅳ组较1μmol·L^(-1)AS-Ⅳ组均降低(P均<0.05)。结论 AS-Ⅳ可能通过抑制活化的小胶质细胞内焦亡通路相关蛋白的表达来发挥其抗炎作用。Objective To investigate the effects of astragaloside IV(AS-IV)on pyroptosis pathway related proteins and its related mechanism in mouse microglia cells.Methods Mouse microglia(BV2)were cultured in vitro.Normal control group was set up,and microglia were activated by thrombin(20 U·mL-1)as the model group.20 U·mL-1 thrombin+three different concentrations of AS-IV(1μmol·L^(-1),5μmol·L^(-1),10μmol·L^(-1))were used as experimental groups.The expressions of pyroptosis related factors(NLRP3,ASC,Caspase-1,GSDMD-N)in BV2 cells were detected by Western blot and Immunofluorescence,and the content of IL-18 and IL-1βin cell culture medium was detected by ELISA.Results The results of Western blot and Immunofluorescence showed that the protein expression of pyroptosis related factors(ASC,NLRP3,Caspase-1,GSDMD-N)in BV2 cells in the thrombin group was significantly higher than that in the control group(P all<0.05).Compared with the thrombin group,the expressions of pyroptosis related factors(ASC,NLRP3,Caspase-1,GSDMD-N)in BV2 cells in different concentrations of AS-IV groups decreased in different degrees(P all<0.05).The reduction degrees of 5 and 10μmol·L^(-1) AS-IV group were more obvious than that of 1μmol·L^(-1) AS-IV group(P all<0.05).ELISA results showed that compared with the control group,the contents of IL-18 and IL-1βin the culture medium of the thrombin group was significantly increased(P all<0.05).Compared with the thrombin group,the contents of IL-18 and IL-1βin the AS-IV groups with different concentrations decreased in different degrees(P all<0.05).The reduction degree of 5 and 10μmol·L^(-1) AS-IV group was more obvious than that of 1μmol·L^(-1) AS-IV group(P all<0.05).Conclusion AS-IV may exert its anti-inflammatory effect by inhibiting the expression of pyroptosis pathway related proteins in activated microglia.

关 键 词：黄芪甲苷 小胶质细胞 细胞焦亡 神经炎症 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]