丹参酮ⅡA通过自噬减轻阿霉素诱导的心肌细胞氧化应激损伤

Tanshinone Ⅱ A attenuates doxorubicin-induced cardiomyocyte oxidative stress injury through autophagy

目的探索丹参酮ⅡA减轻阿霉素诱导的心肌H9C2细胞氧化应激损伤的保护作用和机制。方法将H9C2细胞分为正常组(常规培养),A组(10μg·m L-1丹参酮ⅡA)、B组(1 mmol·L-13-甲基腺嘌呤)、C组(5μmol·L-1阿霉素)、D组(10μg·m L-1丹参酮ⅡA+5μmol·L-1阿霉素)、E组(5μmol·L-1阿霉素+1 mmol·L-13-甲基腺嘌呤)、F组(10μg·m L-1丹参酮ⅡA+5μmol·L-1阿霉素+1 mmol·L-13-甲基腺嘌呤)。7组细胞均孵育24 h。通过细胞计数检测细胞情况,用分光光度法检测细胞丙二醛(MDA)和超氧化物歧化酶(SOD)含量,用免疫荧光检测细胞自噬标志分子LC3。结果正常组和A、B、C、D、E、F组的细胞数量分别为(2.22±0.10),(2.22±0.10),(2.11±0.12),(1.53±0.11),(1.88±0.10),(1.56±0.08)和(1.67±0.08)×106·L-1,MDA分别为(11.25±2.86),(12.12±2.80),(11.33±2.73),(25.07±4.12),(15.07±2.86),(24.57±3.23)和(20.90±2.90)nmol·L-1,SOD分别为(30.95±4.46),(32.72±4.14),(31.12±3.83),(9.63±2.78),(19.62±3.32),(10.45±3.40)和(12.55±3.37)U·m L-1,LC3荧光强度分别为(730.71±139.62),(1375.07±191.05),(374.51±107.04),(1653.24±266.16),(2822.74±391.24),(859.82±113.19)和(1466.32±201.22),正常组与C组比较,D组与C、F组比较,差异均有统计学意义(均P<0.05)。结论丹参酮ⅡA可能通过自噬减轻心肌细胞氧化应激,防治阿霉素所致的心肌细胞损伤。

Objective To investigate the protective effect and mechanism of tanshinoneⅡA on doxorubicin-induced cardiomyocyte oxidative stress injury in myocardial H9C2 cells H9C2.Methods H9C2 cells were divided into normal group(conventional culture),A group(10μg·m L-1tanshinoneⅡA),B group(1 mmol·L-13-methyladenine),C group(5μmol·L-1doxorubicin),D group(10μg·m L-1tanshinoneⅡA+5μmol·L-1doxorubicin),E group(5μmol·L-1doxorubicin+1 mmol·L-13-methyladenine),and F group(10μg·m L-1tanshinoneⅡA+5μmol·L-1doxorubicin+1 mmol·L-13-methyladenine).Cell number was measured by automated cell counter.The concentrations of malondialdehyde(MDA)and superoxide dismutase(SOD)were determined using spectrophotometer.Autophagic marker LC3 wasanalyzed by immunofluorescence assay.Results The number of cells in the normal group and A,B,C,D,E,and Fgroups were(2.22±0.10),(2.22±0.10),(2.11±0.12),(1.53±0.11),(1.88±0.10),(1.56±0.08)and(1.67±0.08)×106·L-1,the contents of MDA were(11.25±2.86),(12.12±2.80),(11.33±2.73),(25.07±4.12),(15.07±2.86),(24.57±3.23)and(20.90±2.90)nmol·L-1,the contents of SOD were(30.95±4.46),(32.72±4.14),(31.12±3.83),(9.63±2.78),(19.62±3.32),(10.45±3.40)and(12.55±3.37)U·m L-1,the LC3 fluorescence intensities were(730.71±139.62),(1375.07±191.05),(374.51±107.04),(1653.24±266.16),(2822.74±391.24),(859.82±113.19)and(1466.32±201.22),the differences were statistically significant between normal group and C group,D group and C or F groups(all P<0.05).Conclusion TanshinoneⅡA could attenuate oxidative stress in H9C2 cardiomyocyte through autophagy,leading to the prevention and treatment of doxorubicin-induced cardiomyocyte injury.