摘要
目的探讨瑞舒伐他汀(RST)在高糖(HG)诱导的人脐静脉内皮细胞(HUVECs)中对Nod样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体激活的调节作用及机制。方法建立HG诱导HUVECs体外模型,细胞分为正常葡萄糖+RST 0(Con)、2(RST 2)、5(RST 5)、10(RST 10)、20(RST 20)、30μmol/L(RST 30)组和HG+RST 0(HG)、2(HG+RST 2)、5(HG+RST 5)、10(HG+RST 10)、20(HG+RST 20)、30μmol/L(HG+RST 30)组。细胞转染实验增加HG+RST10+si-MP活化蛋白激酶(AMPK)组。采用四甲基偶氮唑盐微量酶反应比色法(MTT)法检测细胞活力,H2DCFDA荧光探针检测细胞内活性氧水平,Western blot分析AMPK/NLRP3途径蛋白表达,ELISA法测定上清液IL-1β和IL-18水平。结果HG+RST 5、HG+RST 10组改善HG诱导的细胞损伤,用于后续实验。与HG组比较,HG+RST 5、HG+RST 10组活性氧簇、IL-1β、IL-18水平及NADPH氧化酶4(NOX4)、硫氧还蛋白相互作用蛋白、NLRP3、半胱天冬酶1(Caspase-1)蛋白表达水平降低(P<0.05或P<0.01),p-AMPK/AMPK水平蛋白表达水平升高(P<0.05或P<0.01)。与HG+RST 10组比较,HG+RST10+si-AMPK组NLRP3、Caspase-1蛋白表达水平及IL-1β、IL-18水平升高(P<0.05或P<0.01),p-AMPK/AMPK蛋白表达水平降低(P<0.01)。结论瑞舒伐他汀通过激活AMPK磷酸化和抑制NLRP3炎症小体激活,缓解内皮炎症和氧化应激。
Objective To investigate the effects of Rosuvastatin(RST)on high glucose(HG)-induced NLRP3 inflammasome activation in Human Umbilical Vein Endothelial Cells(HUVECs)and explore the underlying mechanisms.Methods In vitro model of HUVECs induced by HG was established.Cells were divided into normal glucose+RST 0(Con),2(RST 2),5(RST 5),10(RST 10),20(RST 20),30μmol/L(RST 30)group and high glucose+RST 0(HG),2(HG+RST 2),5(HG+RST 5),10(HG+RST 10),20(HG+RST 20),30μmol/L(HG+RST 30)group.HG+RST 10+si-AMPK group was added in cell transfection experiment.Cell viability was detected by MTT assay,ROS level in cells was detected by H2 DCFDA fluorescent probe,AMPK/NLRP3 pathway protein expression was analyzed by Western blot,and IL-1βand IL-18 levels in supernatant were measured by ELISA.Results HG+RST 5,HG+RST 10 improved HG-induced cell damage,which was used in subsequent experiments.Compared with HG group,the levels of ROS,IL-1β,IL-18 and the protein expressions of NOX4,TXNIP,NLRP3 and Caspase-1 decreased(P<0.05 or P<0.01),while the protein expression level of p-AMPK/AMPK protein increased in HG+RST 5 and HG+RST 10 groups(P<0.05 or P<0.01).Compared with HG+RST 10 group,the expressions of NLRP3,Caspase-1 and IL-1β,IL-18 increased(P<0.05 or P<0.01),while the expressions of p-AMPK/AMPK protein decreased in Hg+RST 10+si-AMPK group(P<0.01).Conclusion Rosuvastatin can mitigate endothelial inflammation and oxidative stress by activating AMPK phosphorylation and inhibiting the NLRP3 inflammasome activation.
作者
张寿莉
都渝
ZHANG Shouli;DU Yu(Department of Endocrinology,Qinghai Specialized Hospital of Cardiovascular and Cerebrovascular Diseases,Xining 810012,China)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2021年第10期774-780,共7页
Chinese Journal of Diabetes
基金
青海省医药卫生科技项目(2018-wjzdz-32)。
