摘要
目的通过构建H_(2)O_(2)诱导的H9c2细胞氧化应激模型,观察人参皂苷Rg_(1)对氧化应激的抑制作用,并探讨mi R-499c是否参与人参皂苷Rg_(1)抑制氧化应激的作用机制。方法采用600μmol/L的H_(2)O_(2)诱导H9c2细胞氧化应激模型,给予人参皂苷Rg_(1)预处理24 h,检测乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)活性及丙二醛(malondialdehyde,MDA)、活性氧(reactive oxygen species,ROS)水平和线粒体膜电位(mitochondrial membrane potential,MMP)变化;采用Western blotting检测凋亡相关蛋白表达。采用Lipofiter 3.0将miR-499c转染至H9c2细胞再制备H_(2)O_(2)诱导的氧化应激模型,给予人参皂苷Rg_(1)(40μmol/L)预处理24 h,检测miR-499c表达、ROS水平和MMP变化。结果人参皂苷Rg_(1)显著降低H_(2)O_(2)诱导的H9c2细胞中LDH漏出率、MDA和ROS水平(P<0.05、0.01、0.001),显著升高SOD活性和MMP(P<0.05、0.01、0.001),显著上调B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白表达水平(P<0.05、0.01、0.001),显著下调Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、剪切型半胱氨酸天冬氨酸蛋白酶-3(cleaved cystein-asparate protease-3,cleaved Caspase-3)和cleaved Caspase-9蛋白表达水平(P<0.01、0.001)。人参皂苷Rg_(1)显著增加H9c2细胞中miR-499c表达(P<0.05)。转染miR-499c后,人参皂苷Rg_(1)组ROS水平进一步降低(P<0.05),MMP进一步升高(P<0.05)。结论人参皂苷Rg_(1)通过降低MMP丢失和ROS生成,抑制由H_(2)O_(2)诱导H9c2细胞的氧化应激损伤和细胞凋亡,其作用机制可能与miR-499c相关。
Objective To observe the inhibition effect of ginsenoside Rg_(1) on H_(2)O_(2)-induced oxidative stress in H9c2 cardiomyocytes,and discuss whether miR-499c involved in the mechanism of ginsenoside Rg_(1) inhibition of oxidative stress.Methods The oxidative stress model of H9c2 cardiomyocytes was induced by 600μmol/L H_(2)O_(2),ginsenoside Rg_(1) were pretreated for 24 h.Lactate dehydrogenase(LDH),superoxide dismutase(SOD)activities and malondialdehyde(MDA),reactive oxygen species(ROS)levels and mitochondrial membrane potential(MMP)changes were detected;Western blotting was used to detect the expressions of apoptosisrelated proteins.Lipofiter 3.0 was used to transfect miR-499c into H9c2 cells,and then H_(2)O_(2)-induced oxidative stress model was established,ginsenoside Rg_(1)(40μmol/L)was pretreated for 24 h,and miR-499c expression,ROS level and MMP changes were detected.Results Ginsenoside Rg_(1) significantly decreased the leakage rate of LDH,MDA and ROS levels in H9c2 cells induced by H_(2)O_(2)(P<0.05,0.01,0.001),significantly increased SOD activity and MMP(P<0.05,0.01,0.001),significantly increased B-cell lymphoma-2(Bcl-2)protein expression level(P<0.05,0.01,0.001),significantly decreased Bcl-2 associated X protein(Bax),cleaved cysteine aspartate protease-3(cleaved Caspase-3)and cleaved Caspase-9 protein expression levels(P<0.01,0.001).Ginsenoside Rg_(1) significantly increased the expression of miR-499c in H9c2 cells(P<0.05).After transfection with miR-499c,the level of ROS in ginsenoside Rg_(1) group was further decreased(P<0.05),and MMP was further increased(P<0.05).Conclusion Ginsenoside Rg_(1) can inhibit oxidative stress injury and apoptosis of H9c2 cells induced by H_(2)O_(2) by reducing MMP loss and ROS production,and its mechanism may be related to miR-499c.
作者
石洪洋
董慧
刘嘉
崔笑天
郭毅
洪兰
SHI Hong-yang;DONG Hui;LIU Jia;CUI Xiao-tian;GUO Yi;HONG Lan(Department of Food and Biological Sciences,School of Agriculture,Yanbian University,Yanji 133002,China;Department of Physiology and pathophysiology,Medical School,Yanbian University,Yanji 133002,China;Affiliated Hospital of Changchun University of Chinese Medicine,Changchun 130000,China)
出处
《中草药》
CAS
CSCD
北大核心
2023年第24期8117-8126,共10页
Chinese Traditional and Herbal Drugs
基金
吉林省教育厅科学技术研究项目(JJKH20220533KJ)。
