胡黄连苷Ⅱ减轻阿霉素诱发H9C2心肌细胞损伤的潜在机制研究  

Study on the potential mechanism of picrosideⅡin reducing adriamycin-induced H9C2 cell injury

作　　者：陈泺帆 赵金丹 饶红[1] CHEN Luofan;ZHAO Jindan;RAO Hong(Center for Cardiovascular Clinical Medicine,Liyuan Hospital Affiliated to Tongji Medical College of Huazhong University Science and Technology,Wuhan,Hubei 430077,China)

机构地区：[1]华中科技大学同济医学院附属梨园医院心血管临床医学中心,武汉430077

出　　处：《重庆医学》2023年第19期2899-2904,2911,共7页Chongqing medicine

摘　　要：目的研究胡黄连苷Ⅱ(Pic)减轻阿霉素(Dox)诱发H9C2心肌细胞损伤的潜在机制。方法采用Dox(1μmol/L)处理H9C2心肌细胞24 h以构建Dox诱发心肌损伤的细胞模型。采用CCK-8法检测不同浓度Pic对细胞活性的影响,以确定Pic最佳的干预浓度。将细胞汇合度达到60%~70%的H9C2心肌细胞随机分为对照组(Con组)、阿霉素组(Dox组)和治疗组(Dox+Pic组)。采用乳酸脱氢酶(LDH)水平评价Dox对细胞的损伤;采用膜联蛋白V(Annexin V)-异硫氰酸荧光素(FITC)/碘化丙啶(PI)染色和TUNEL染色评估细胞凋亡率;使用JC-1染色判定Dox和Pic对H9C2心肌细胞线粒体膜电位(MMP)的影响;通过Western blot检测各组凋亡相关蛋白(BAX、BCL2、C-caspase-3)的表达情况;使用去乙酰化酶沉默信息调节因子1(SIRT1)选择性抑制剂(SIRT1-IN-1)研究Pic是否通过调控SIRT1来发挥对Dox处理细胞的保护作用。结果Dox能明显降低H9C2心肌细胞活性,而Pic则可抑制这一过程,与Con组比较,Dox+Pic组细胞培养基中的LDH水平明显降低。Annexin V-FITC/PI染色和TUNEL染色均表明Pic可以抑制Dox导致的细胞凋亡。JC-1染色结果显示,与Dox组比较,Dox+Pic组细胞的MMP降低。Western blot检测表明,Pic能够抑制促凋亡蛋白BAX、C-caspase-3表达,促进抗凋亡蛋白BCL2表达。Dox组细胞SIRT1蛋白表达降低,但Pic能够上调SIRT1表达,而SIRT1-IN-I(2μmol/L)逆转了Pic的抗凋亡作用。结论Pic可以通过激活SIRT1蛋白表达抑制Dox引起的细胞损伤。Objective To investigate whether PicrosideⅡ(Pic)could inhibit the damage of H9C2 cell caused by doxorubicin(Dox)and its potential mechanisms.Methods H9C2 cardiomyocytes were treated with Dox(1μmol/L)for 24 hours to construct a cellular model of Dox-induced myocardial injury.Firstly,the effect of different concentrations of Pic on cell activity was detected using the Cell Counting Kit-8(CCK-8)method to determine the optimal concentration of Pic for intervention.Subsequently,H9C2 cells with 60%-70%confluence were then randomly divided into three groups:the control group(Con),the Dox group and the treatment group(Dox+Pic).Cell damage by Dox was evaluated using lactate dehydrogenase(LDH)levels;cellular apoptotic rate was assessed using Annexin V-FITC/PI staining and TUNEL staining.The effects of Dox and Pic on mitochondrial membrane potential(MMP)of H9C2 cells were determined using JC-1 dye;the expression of apoptosis-related proteins(BAX,BCL2,C-caspase-3)in each group was detected by Western blotting assay.A selective inhibitor of deacetylase silent information reglator 1(SIRT1-IN-1)was employed to explore whether Pic could exert a protective effect against Dox-treated cells by regulating SIRT1.Results Dox significantly reduced H9C2 cell activity,while Pic inhibited this process.Compared with the Con group and LDH levels in cell culture media in the Dox+Pic group were significantly reduced.Annexin V-FITC/PI staining and TUNEL staining both showed that Pic couldreduce apoptosis caused by Dox,and JC-1 staining showed that the mitochondrial membrane potential(MMP)of cells in the Dox+Pic group was significantly reduced compared with the Dox group.Western blotting assay showed that Pic could inhibit the expression of pro-apoptotic proteins(BAX,Cleaved caspase-3)and promote the expression of anti-apoptotic protein BCL2.Mechanistically,SIRT1 protein expression was reduced in the Dox group cells,yet Pic could upregulate SIRT1 expression,and SIRT1-IN-I(2μmol/L)reversed the anti-apoptotic effect of Pic.Conclusion Pic can

关 键 词：胡黄连苷Ⅱ 阿霉素 心肌细胞凋亡 H9C2心肌细胞损伤 

分 类 号：R542.2[医药卫生—心血管疾病]