海南医学院学报2024，Vol.30Issue(7)：481-488,8.DOI:10.13210/j.cnki.jhmu.20240030.001
黄芩苷通过ROS依赖性调节RhoA/ROCK通路保护氯化钴诱导心肌细胞损伤的实验研究
Experimental study of baicalin alleviating hypoxia-induced H9c2 cell injury through ROS-dependent RhoA/ROCK pathway
摘要
Abstract
Objective:To investigate the mechanism of Baicalin on the hypoxic injury of H9c2 cardiomyocytes induced by co-balt chloride(CoCl2).Methods:Cobalt chloride(CoCl2)was used to establish a hypoxic injury model in cardiomyocytes,and dif-ferent concentrations of Baicalin were added for culturing,respectively.The normoxic culture was set as the control group,and the cobalt chloride cultured H9c2 cardiomyocytes were set as the CoCl2 group.The hypoxic H9c2 cardiomyocytes pretreated with Ba-icalin and/or Y27632(Rho kinase inhibitor)were set as the CoCl2+Baicalin group,CoCl2+Y27632 group and CoCl2+Baicalin+ Y27632 group.Cell proliferation was assessed using the cytotoxicity assay kit(CCK8)assay,and reactive oxygen species(ROS)levels were detected using a fluorescence method.The activity of superoxide dismutase(SOD)in cardiomyocytes was detected by WST-8.The concentration of propylene glycol(MDA)was determined by TBA method.Meanwhile,the expression of RhoA,ROCK1,ROCK2,TNF-α and IL-1β were analyzed by Western blot as well.Results:After treatment of H9c2 cells for 24 h,1 000 μmol/L CoCl2 and 75 μmol/L Baicalin showed better cell viability in the treatment of hypoxic injury of cardiomyocytes.The cell activity of the CoCl2 group was significantly lower than that of the control group,and the pre-protective effect of Baicalin on hypoxia-induced cells could significantly increase the cell activity.Compared with the control group,ROS expression level and MDA content in cardiomyocytes of the CoCl2 group increased,protein expression levels of RhoA,ROCK1,ROCK2,TNF-α and IL-1β up-regulated,and SOD activity decreased.Compared with the CoCl2 group,CoCl2+Baicalin could inhibit the expression of ROS and MDA content,up-regulate SOD activity,and down-regulate the expression levels of RhoA,ROCK1,ROCK2,TNF-α and IL-1β,while adding Y27632(Rho kinase inhibitor)significantly enhanced the protective effect of Baicalin.Conclusion:Ba-icalin could alleviate inflammatory and oxidative stress responses to CoCl2-induced injury in H9c2 cardiomyocytes,and its mecha-nism might be related to the inhibition of ROS-dependent RhoA/ROCK pathway.关键词
黄芩苷/RhoA/Rho相关激酶(ROCK)信号通路/缺氧损伤模型Key words
Baicalin/RhoA/Rho-related kinase(ROCK)signaling pathway/Hypoxia injury model分类
医药卫生引用本文复制引用
沈艳玲,刘承红,王世魁,徐尧,张云波,顾申红..黄芩苷通过ROS依赖性调节RhoA/ROCK通路保护氯化钴诱导心肌细胞损伤的实验研究[J].海南医学院学报,2024,30(7):481-488,8.基金项目
海南省重点研发项目(EPC‑MVs富集miR‑126作用于靶基因抑制RhoA/ROCK通路对高血压肾病作用及机制、ZDYF2022SHFZ101)This study was supported by Key R&D Projects in Hainan Province(EPC-MVs enrichment of miR-126 acting on target genes,inhibition of RhoA/ROCK pathway on hypertensive nephropathy and its mechanism,ZDYF2022SHFZ101) （EPC‑MVs富集miR‑126作用于靶基因抑制RhoA/ROCK通路对高血压肾病作用及机制、ZDYF2022SHFZ101）
