摘要
目的探讨黄芩苷对阿霉素(Dox)诱导的H9c2细胞毒性的影响及内在机制。方法采用50μmol/L黄芩苷预处理H9c2细胞24 h,然后1μmol/L Dox处理H9c2细胞24 h,建立体外Dox心肌毒性模型。采用CCK8法检测细胞活力;收集细胞上清检测各组心肌损伤标志物乳酸脱氢酶(LDH)、心肌肌钙蛋白I(cTnI)和肌酸激酶同工酶(CK-MB)的水平以及氧化应激相关指标超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)的水平;使用DHE试剂盒检测各组活性氧(ROS)的含量;使用TUNEL染色检测各组细胞凋亡水平,RT-qPCR和Western blot实验用于检测氧化应激和凋亡相关分子的表达水平。结果与Dox组相比,黄芩苷能提高H9c2细胞活力,降低LDH、cTnI、CK-MB水平;DHE染色显示黄芩苷能减少ROS的生成,增加SOD、GSH-Px的活性,降低MDA的含量;TUNEL染色结果显示黄芩苷能减少阳性细胞数量;RT-qPCR和Western blot检测显示黄芩苷能上调Nrf2、HO-1、SOD2、Bcl-2的表达,降低Cleaved-caspase 3和Bax的表达。然而,Nrf2的特异性抑制剂ML385可逆转黄芩苷引起的上述变化。结论黄芩苷通过上调Nrf2/HO-1信号通路减轻氧化应激和凋亡,减轻Dox诱导的H9c2细胞毒性。
Objective To investigate the effect and underlying mechanism of baicalin on doxorubicin(Dox)-induced H9c2 cell toxicity.Methods H9c2 cells were pretreated with 50μmol/L baicalin for 24 h,followed by treatment with 1μmol/L Dox for 24 h to establish an invitro model of Dox-induced myocardial toxicity.Cell viability was assessed using the CCK8 assay.The levels of lactate dehydrogenase(LDH),cardiac troponin I(cTnI),creatine kinase isoenzyme(CK-MB),as well as oxidative stress-related indicators such as superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and malondialdehyde(MDA)were measured in the cell supernatant of each group.Reactive oxygen species(ROS)content was determined using the DHE assay kit.TUNEL staining was employed to assess cell apoptosis levels in each group.Additionally,RT-qPCR and Western blot experiments were conducted to measure the expression levels of oxidative stress and apoptosis-related molecules.Results Baicalin demonstrated the ability to enhance H9c2 cell viability and decrease LDH,cTnI,and CK-MB levels compared to the Dox group.DHE staining indicated that baicalin reduced ROS generation,increased SOD and GSH-Px activity,and decreased MDA content.TUNEL staining results revealed a reduction in the number of positive cells with baicalin treatment.RT-qPCR and Western blot analysis showed that baicalin upregulated the expression of Nrf2,HO-1,SOD2,and Bcl-2,while downregulating the expression of Cleaved-caspase 3 and Bax.However,ML385,a specific inhibitor of Nrf2,reversed the above changes induced by baicalin.Conclusion Baicalin alleviates oxidative stress and apoptosis by upregulating the Nrf2/HO-1 signaling pathway,thereby mitigating Dox-induced H9c2 cell toxicity.
作者
李登科
张伟
黄从新
LI Dengke;ZHANG Wei;HUANG Congxin(Department of Cardiology,Renmin Hospital of Wuhan University,Cardiovascular Research Institute,Wuhan University,Hubei Key Laboratory of Cardiology,Wuhan 430060,Hubei,China)
出处
《心血管病学进展》
CAS
2024年第5期457-465,共9页
Advances in Cardiovascular Diseases
基金
湖北省技术创新专项(2016ACA153)。
