摘要
目的 探讨黄芪甲苷对高糖诱导的心肌细胞焦亡的影响。方法 体外培养H9c2细胞,分为对照组(5.5 mmol/L葡萄糖)、高糖组(33.3 mmol/L葡萄糖)、黄芪甲苷组(33.3 mmol/L葡萄糖+100μmol/L黄芪甲苷)、NLRP3抑制剂组(33.3 mmol/L葡萄糖+1μmol/L MCC950)。细胞计数试剂盒8(CCK-8)检测H9c2细胞活性,乳酸脱氢酶(LDH)试剂盒检测细胞上清液中LDH含量,超氧化物阴离子荧光探针(DHE)检测细胞内活性氧(ROS)水平,实时荧光定量聚合酶链式反应(RT-qPCR)及免疫印迹法(Western blot)检测焦亡相关基因mRNA和蛋白表达水平,免疫荧光法检测NLRP3荧光强度,酶联免疫吸附法(ELISA)检测细胞上清液中炎症因子水平。结果 黄芪甲苷浓度为100μmol/L时可改善高糖诱导的细胞活力下降(P<0.01),减少LDH释放(P<0.01)。与对照组相比,高糖组ROS水平升高(P<0.01),细胞焦亡相关分子mRNA和蛋白表达上调(均P<0.01),NLRP3荧光强度增加(P<0.01),细胞上清液中炎症因子水平升高(P<0.01);与高糖组相比,黄芪甲苷组和抑制剂组ROS水平降低(P<0.01),细胞焦亡相关分子mRNA和蛋白表达下调(P<0.05或P<0.01),NLRP3荧光强度减少(P<0.01),细胞上清液中炎症因子水平降低(P<0.05或P<0.01)。结论 黄芪甲苷通过抑制NLRP3/Caspase-1信号通路,抑制细胞焦亡,对高糖诱导的心肌细胞损伤发挥保护作用。同时,它可以提高细胞模型的抗炎性和抗氧化性。
Objective To investigate the effect of AstragalosideⅣon high glucose-induced cardiomyocyte pyroptosis.Methods H9c2 cells were cultured in vitro and divided into control group(5.5 mmol/L glucose),high glucose group(33.3 mmol/L glucose),AstragalosideⅣgroup(33.3 mmol/L glucose+100μmol/L AstragalosideⅣ),and NLRP3 inhibitor group(33.3 mmol/L glucose+1μmol/L MCC950).Cell counting kit 8(CCK-8)was used to detect the activity of H9c2 cells.Lactate dehydrogenase(LDH)kit was used to detect the content of LDH in cell supernatant.Superoxide anion fluorescent probe(DHE)was used to detect the level of intracellular reactive oxygen species(ROS).Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression levels of pyroptosis-related genes.Immunofluorescence was used to detect the fluorescence intensity of NLRP3.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of inflammatory factors in cell supernatant.Results When the concentration of AstragalosideⅣwas 100μmol/L,it could significantly inhibit the decrease of cardiomyocyte viability induced by high glucose(P<0.01)and reduce LDH release(P<0.01).Compared with the control group,the level of ROS was increased(P<0.01),the mRNA and protein expressions of pyroptosis-related molecules were up-regulated(P<0.01 for all),the fluorescence intensity of NLRP3 was increased(P<0.01),and the levels of inflammatory factors in the cell supernatant were increased in the high glucose group(P<0.01).Compared with the high glucose group,the ROS level was decreased(P<0.01),the mRNA and protein expressions of pyroptosis-related molecules were down-regulated(P<0.05 or P<0.01),the fluorescence intensity of NLRP3 was decreased(P<0.01),and the levels of inflammatory factors in cell supernatant were decreased(P<0.05 or P<0.01)in AstragalosideⅣgroup and inhibitor group.Conclusions AstragalosideⅣplays a protective role in high glucose-induced cardiomyocyte injury by inhibiting NLRP3/Caspase-1 signali
作者
秦依然
张志强
陈学恒
尉希清
张金国
Qin Yiran;Zhang Zhiqiang;Chen Xueheng;Wei Xiqing;Zhang Jinguo(Cheeloo College of Medicine,Shandong University,Jinan 250012,China;Clinical Medical School,Jining Medical University,Jining 272067,China;Department of Cardiology,Affiliated Hospital of Jining Medical University,Jining 272029,China)
出处
《中华老年医学杂志》
CAS
CSCD
北大核心
2023年第3期341-346,共6页
Chinese Journal of Geriatrics
基金
山东省自然科学基金(ZR202112020064)
山东省中医药科技发展计划项目(2019-0481)
济宁市重点研发计划项目(2021YXNS069)。
关键词
黄芪甲苷
高糖
细胞焦亡
心肌细胞
AstragalosideⅣ
High glucose
Pyroptosis
Cardiomyocytes
