白果内酯对Aβ25-35诱导HT22细胞损伤及NLRP-1炎症小体激活的影响  

Effects of bilobalide on Aβ-induced HT22 cell injury and the activation of NLRP-1 inflammasome

作　　者：黄茸茸[1] 陆松侠 许燕 李维祖[2] HUANG Rong-rong;LU Song-xia;XU Yan;LI Wei-zu(Department of Pharmacology,Anhui Xinhua University,Hefei 230088,China;Department of Pharmacology/Key Laboratory of Antiinflammatory and Immunopharmacology/Ministry of Education Key Laboratory for Research and Development of Chinese Medicine/Level 3 Laborotory of State Administration of Traditional Chinese Medicine,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽新华学院药学院药理教研室,安徽合肥230088 [2]安徽医科大学药理学教研室,抗炎免疫药理学教育部重点实验室,国家中医药管理局中药药理三级实验室,安徽合肥230032

出　　处：《中成药》2020年第9期2317-2323,共7页Chinese Traditional Patent Medicine

基　　金：安徽省教育厅高校自然科学研究重点项目(KJ2015A298);安徽省教育厅质量工程项目(2018mooc438)。

摘　　要：目的研究白果内酯对Aβ25-35诱导HT22细胞损伤及NLRP-1炎症小体激活的影响。方法Aβ25-35(1μmol/L)诱导HT22建立细胞损伤模型,同时白果内酯(6、12、24μmol/L)和NADPH氧化酶抑制剂(DPI,15μmol/L)处理24 h,采用CCK-8法检测细胞活力,Annexin V-FITC/PI双染法检测神经元细胞凋亡率,流式法(DCFH-DA荧光探针)测定细胞内ROS生成,ELISA法测定IL-1β、IL-6、TNF-α水平,Western blot法检测NLRP-1、ASC、caspase-1的蛋白表达。结果与对照组比较,模型组细胞活力下降(P<0.01),神经元细胞凋亡率提高(P<0.01),同时,ROS的生成、细胞因子IL-1β、IL-6、TNF-α水平及NLRP-1、ASC、caspase-1的蛋白表达均增多(P<0.01);与模型组比较,白果内酯(12、24μmol/L)组能提高细胞活力,减少ROS生成,降低IL-1β、IL-6、TNF-α水平与神经元细胞凋亡率(P<0.05,P<0.01),白果内酯(24μmol/L)组能抑制细胞NLRP-1、ASC及caspase-1的蛋白表达(P<0.01)。结论白果内酯对Aβ25-35诱导的小鼠海马神经元HT22细胞损伤具有较好的保护作用,其机制可能与抑制NADPH氧化酶-ROS介导的NLRP-1炎症小体激活有关。AIM To investigate the effects of bilobalide on Aβ25-35-induced HT22 cell injury and the activation of NLRP-1 inflammasome.METHODS Cell damage models built upon HT22 cells by Aβ25-35 induction were subsequently exposed to 24 h treatments of bilobalide(6,12,24 mol/L)and NADPH oxidase inhibitor(DPI,15 mol/L)respectively.The activity of HT22 cells and neuronal apoptosis rate were detected by CCK-8 method and Annexin V-FITC/PI double staining,the production of reactive oxygen species(ROS)was examined by DCFH-DA fluorescent probe,the levels of IL-1β,IL-6 and TNF-αwere determined by ELISA,and the expression of NLRP-1,ASC and caspase-1 protein in neurons were identified by Western blot.RESULTS Compared with the control group,the model group was observed with decreased cell activity and increase neuronal apoptosis rate(P<0.01),more ROS production,and increased levels of cytokines IL-1β,IL-6,TNF-α,and stronger expression of NLRP-1,ASC and caspase-1 protein(P<0.01).Bilobalide at concentrations of 12μmol/L and 24μmol/L improved cell viability,reduced ROS production,and lowered IL-1β,IL-6,TNF-αlevels and neuronal cell apoptosis rate(P<0.05,P<0.01).Bilobalide at concentration of 24μmol/L inhibited the protein expression of NLRP-1,ASC and caspase-1(P<0.01).CONCLUSION Bilobalide’s protective effect on Aβ25-35-induced HT22 cells damage may be associated with its inhibition to NADPH oxidase-ROS-mediated activation of NLRP-1 inflammasome.

关 键 词：白果内酯 阿尔茨海默病 AΒ25-35 ROS NLRP-1 ASC CASPASE-1 

分 类 号：R285.5[医药卫生—中药学]