黄芪多糖保护心肌细胞对抗H_(2)O_(2)诱导的细胞凋亡相关机制  

The Mechanisms of Astragalus Polysaccharide for Protecting Cardiomyocytes against H_(2)O_(2)-induced Apoptosis

作　　者：吴晟洁 万四园 王哲文 杨诗雨 王易慧 国海东[1] 李涵 WU Shengjie;WAN Siyuan;WANG Zhewen;YANG Shiyu;WANG Yihui;GUO Haidong;LI Han(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学,上海201203

出　　处：《中西医结合心脑血管病杂志》2025年第6期841-847,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金项目(No.82174120);上海市自然科学基金项目(No.21ZR1463100);上海市优秀学术带头人计划项目(No.22XD1423400)。

摘　　要：目的:探讨黄芪多糖保护大鼠心肌细胞(H9c2)对抗过氧化氢(H_(2)O_(2))诱导的细胞凋亡及机制。方法:体外培养H9c2细胞,经黄芪多糖预处理后,通过H_(2)O_(2)模拟体外氧化应激模型,采用细胞计数试剂盒(CCK-8)、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)分别检测黄芪多糖对H9c2细胞增殖活性的影响和对抗H_(2)O_(2)诱导产生的氧化应激损伤的影响。通过蛋白免疫印迹法(Western Blot)检测磷酸化蛋白激酶B(p-AKT)、磷酸化细胞外调节蛋白激酶1/2(p-ERK1/2)水平,探讨黄芪多糖在体外对H9c2细胞的保护作用机制。结果:100μg/mL黄芪多糖可显著促进H9c2细胞的增殖活性。350μmol/L H_(2)O_(2)导致H9c2细胞凋亡率约为50%。采用350μmol/L的H_(2)O_(2)建立体外氧化应激模型,CCK-8活性检测显示,100μg/mL的黄芪多糖保护H9c2细胞对抗H_(2)O_(2)诱导产生的氧化应激损伤的作用显著。Western Blot检测结果显示,与Control组、H_(2)O_(2)组比较,黄芪多糖+H_(2)O_(2)组激活了p-AKT及p-ERK1/2蛋白表达。磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)及丝裂原活化蛋白激酶(MAPK)/细胞外调节蛋白激酶(ERK)信号通路抑制剂LY294002及U0126抑制了黄芪多糖对信号通路的激活作用。利用黄芪多糖和H_(2)O_(2)处理H9c2细胞后,TUNEL检测显示,黄芪多糖+H_(2)O_(2)组细胞凋亡数目减少(P<0.05)。结论:黄芪多糖可保护H9c2细胞对抗H_(2)O_(2)诱导产生的氧化应激损伤,其机制可能与激活PI3K/AKT、MAPK/ERK信号通路有关。Objective:To investigate the protective effect of Astragalus polysaccharide on rat cardiomyocytes(H9c2)against hydrogen peroxide(H_(2)O_(2))-induced apoptosis.Methods:H9c2 cells were cultured in vitro.After pretreatment with Astragalus polysaccharide,the oxidative stress model was simulated by H_(2)O_(2).The effects of Astragalus polysaccharides on the proliferation of H9c2 cells and their resistance to oxidative stress induced by H_(2)O_(2)were detected by cell counting kit(CCK-8)and terminal deoxynucleotide transferase mediated dUTP notch end labeling(TUNEL).The levels of phosphorylated protein kinase B(p-AKT)and phosphorylated extracellular regulatory protein kinase 1/2(p-ERK1/2)were detected by Western Blot to explore the protective effect of Astragalus polysaccharide on H9c2 cells in vitro.Results:The 100μg/mL Astragalus polysaccharide could significantly promote the proliferation activity of H9c2 cells.The apoptosis rate of H9c2 cells was about 50%due to 350μmol/L H_(2)O_(2).In vitro oxidative stress model was established using 350μmol/L H_(2)O_(2).CCK-8 activity detection showed that 100μg/mL of Astragalus polysaccharide showed a significant protective effect on H9c2 cells against oxidative stress damage induced by H_(2)O_(2).The results of Western Blot analysis showed that compared with Control group and H_(2)O_(2)group,Astragalus polysaccharide+H_(2)O_(2)group p-AKT and p-ERK1/2 protein expression activated.Phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)and mitogen-activated protein kinase(MAPK)/extracellular regulatory protein kinase(ERK)signaling pathway inhibitors LY294002 and U0126 inhibited the activation of the signaling pathway by Astragalus polysaccharides.After the treatment of H9c2 cells with Astragalus polysaccharide and H_(2)O_(2),TUNEL detection showed that the number of apoptosis of H9c2 cells in the Astragalus polysaccharide+H_(2)O_(2)group decreased(P<0.05).Conclusion:Astragalus polysaccharide could protect H9c2 cells against oxidative stress induced by H_(2)O_(2),and its me

关 键 词：H9C2细胞 黄芪多糖 氧化应激 细胞凋亡 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 丝裂原活化蛋白激酶/细胞外调节蛋白激酶信号通路 实验研究 

分 类 号：R285[医药卫生—中药学]