小白菊内酯对多柔比星诱导的H9c2心肌细胞损伤保护作用的相关机制研究  

Study on the Protective Effects and Mechanisms of Parthenolide Against Doxorubicin-Induced H9c2 Cardiomyocyte Injury

作　　者：郭红豆 蒋凌 罗顺祥 陈甘潇 林佳仪 林奕帆 徐尚华 GUO Hongdou;JIANG Ling;LUO Shunxiang;CHEN Ganxiao;LIN Jiayi;LIN Yifan;XU Shanghua(Department of Cardiology,Affiliated Nanping First Hospital,Fujian Medical University,Nanping 353000,Fujian,China;Department of Cardiology,Wuwei People’s Hospital,Wuwei 733000,Gansu,China)

机构地区：[1]福建医科大学附属南平第一医院心血管内科,福建南平353000 [2]武威市人民医院心血管内科,甘肃武威733000

出　　处：《心血管病学进展》2024年第7期666-672,共7页Advances in Cardiovascular Diseases

基　　金：福建省自然科学基金(2019J01610);福建省科技创新联合基金项目(2021Y9035)。

摘　　要：目的探索小白菊内酯(PTL)对多柔比星(DOX)诱导的心肌细胞损伤的保护作用机制。方法培养H9c2大鼠心肌细胞,应用CCK-8法确定DOX和PTL的最佳作用浓度。将H9c2大鼠心肌细胞分为对照组、PTL组、DOX组及DOX+PTL组进行相应处理,DCFH-DA染色流式细胞仪检测细胞内活性氧水平,Annexin V-FITC/PI染色流式细胞仪检测细胞内凋亡比例,蛋白质印迹法检测核转录因子红系2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、B细胞淋巴瘤-2相关x蛋白(Bax)和B细胞淋巴瘤-2(Bcl-2)蛋白的表达。结果检测0.5~5.0μmol/L浓度梯度的DOX处理大鼠H9c2心肌细胞24 h后,细胞活力下降,并且呈现剂量依赖性(P<0.05),其中1.0μmol/L DOX可引起H9c2心肌细胞活力降至53.0%±0.4%。5.0μmol/L PTL预处理H9c2心肌细胞3 h后可显著增加DOX引起的细胞活力(P<0.05)。与对照组相比,在DOX组的H9c2心肌细胞中Nrf2、HO-1及Bcl-2蛋白表达显著降低,Bax表达显著增加(P<0.05),氧化应激水平、细胞凋亡比例增多(P<0.05);与DOX组相比,PTL预处理显著增加了DOX处理的H9c2心肌细胞中Nrf2、HO-1、Bcl-2蛋白表达,降低Bax表达(P<0.05),并降低氧化应激水平、细胞凋亡率(P<0.05)。结论PTL缓解DOX引起的H9c2心肌细胞损伤,可能与激活Nrf2/HO-1通路、抑制Bcl-2/Bax通路有关。Objective To investigate the protective mechanism of parthenolide(PTL)against doxorubicin(DOX)-induced cardiomyocyte injury.Methods Rat H9c2 cardiomyocytes were cultured,and then the optimal concentration of DOX and PTL was determined by CCK-8 assay.Rat H9c2 cardiomyocytes were randomly divided into control group,PTL group,DOX group and DOX+PTL group.Intracellular reactive oxygen species levels were detected with DCFH-DA using a flow cytometer and the apoptotic cells were measured with Annexin V-FITC/PI staining kit by flow cytometer.Western blot was employed to detect the expressions of nuclear factor-erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),B-cell lymphoma-2-associated X protein(Bax)and B-cell lymphoma-2(Bcl-2).Results The cell viability of H9c2 cardiomyocytes exposed to 0.5~5.0μmol/L DOX for 24 h decreased in a dose-dependent effect(P<0.05),and 1.0μmol/L DOX could reduce the viability of H9c2 cardiomyocytes to 53.0%±0.4%.The cell viability of H9c2 cardiomyocytes pretreated with 5.0μmol/L PTL for 3 h was significantly increased after exposure to DOX(P<0.05).The protein expression levels of Nrf2,HO-1 and Bcl-2 in DOX group were significantly decreased comparing with the control group,while the proapoptotic protein Bax was significantly increased(P<0.05),and the level of oxidative stress and the proportion of apoptosis were increased(P<0.05).Contrasted with DOX group,pretreatment with PTL could significantly increase the expression levels of Nrf2,HO-1 and Bcl-2 protein,and reduce the expression level of Bax(P<0.05),and decrease the degree of oxidative stress and apoptosis rate of DOX-induced H9c2 cardiomyocytes(P<0.05).Conclusion PTL alleviates DOX-induced injury of H9c2 cardiomyocytes which was associated with activation of Nrf2/HO-1 pathway and inhibition of Bcl-2/Bax signaling pathway.

关 键 词：小白菊内酯 多柔比星 H9C2心肌细胞 Nrf2/HO-1信号通路 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]