加味丹参饮含药血清对H9C2心肌细胞缺氧/复氧损伤的保护作用

Protective Effect of Jiawei Danshen Decoction Containing Serum on Hypoxia/reoxygenation Injury of H9C2 Cardiomyocytes

目的:从Nrf2–ARE信号通路探讨加味丹参饮对H9C2心肌细胞缺氧/复氧(H/R)损伤的保护机制。方法:制备SD大鼠含药血清及空白血清,以不同浓度加味丹参饮含药血清干预正常及H/R损伤H9C2心肌细胞,用CCK–8法筛选加味丹参饮的实验剂量。体外培养H9C2心肌细胞建立缺氧/复氧损伤模型,将H9C2心肌细胞随机分为6组:正常组、H/R组、空白血清组(BS组)、加味丹参饮含药血清组(JDD组)、JDD+Nrf2抑制剂组(JDD+ML385组)、活性氧(ROS)清除剂组(NAC组),比较各组心肌细胞间形态学变化的差别。结果:选择15%浓度的加味丹参饮含药血清为工作浓度血清。对比正常组,H/R组倒置显微镜下细胞变性坏死程度增加,JDD+ML385组、BS组与H/R组在细胞形态的表现上相似,而JDD组及NAC组干预后,H9C2心肌细胞的病理状态均有明显改善。结论:加味丹参饮抗心肌缺血再灌注损伤(MIRI)的保护作用可能通过活化Nrf2–ARE通路减轻活性氧(ROS)介导的氧化应激损伤来实现。

Objective To explore the protective effect and its mechanism of Jiawei Danshen decoction on hypoxia/reoxygenation(H/R)injured H9C2 cardiomyocytes from Nrf2–ARE signaling pathway.Methods Drug-containing serum and blank serum of SD rats were prepared,and different concentrations of drug-containing serum of Jiawei Danshen decoction were used to interfere with normal and H/R injury of H9C2 myocardial cells.The experimental dose of Jiawei Danshen decoction was screened by CCK-8 method.H9C2 cardiomyocytes were cultured in vitro to establish H/R injury model.H9C2 cardiomyocytes were randomly divided into six groups:normal group,H/R group,blank serum group(BS group),Jiawei Danshen decoction containing serum group(JDD group),JDD+Nrf2 inhibitor group(JDD+ML385 group)and reactive oxygen species(ROS)scavenger group(NAC group).The morphological changes of cardiomyocytes in each group were compared.Results The serum containing 15%concentration of Jiawei Danshen decoction was selected as the working concentration serum.Compared with the normal group,the degree of cell degeneration and necrosis in H/R group increased under the inverted microscope.The cell morphology of JDD+ML385 group,BS group and H/R group were similar,while the pathological state of H9C2 cardiomyocytes was significantly improved after intervention in JDD group and NAC group.Conclusion The protective effect of Jiawei Danshen decoction against myocardial ischemia-reperfusion injury(MIRI)may be realized by activating NrF2-ARE pathway to reduce oxidative stress injury mediated by ROS.