丹红注射液预处理对乳鼠心肌细胞缺氧/复氧损伤的保护作用及其机制

Study of preconditioning with Danhong injection protects cultured neonatal rat myocardial cells against anoxia/reoxygenation injury

目的探讨丹红注射液(DH)预处理对心肌细胞缺氧/复氧损伤的保护作用及其相关机制。方法建立乳鼠心肌细胞缺氧/复氧损伤模型,分离培养SD乳鼠心肌细胞,在培养3d后随机分为5组(每组实验重复6次):正常对照(control)组、缺氧/复氧(A/R)组,丹红注射液低剂量(终浓度为0.2μmol/ml)(DHL+A/R)组,丹红注射液中剂量(终浓度为0.6μmol/ml)(DHM+A/R)组,丹红注射液高剂量(终浓度为1.8μmol/ml)(DHH+A/R)组。测定各组心肌细胞存活率,培养液中乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)的含量,以及细胞内ATP水平。结果与对照组比较,A/R组心肌细胞存活率、细胞培养液中SOD活性和细胞内ATP水平均明显降低(均P<0.01),而培养液中LDH水平明显增加(P<0.01)。与A/R组比较,DH+A/R各剂量组心肌细胞存活率、细胞内SOD含量和细胞内ATP水平均明显增加(均P<0.01),而培养液中LDH水平均明显降低(P<0.01);其中又以DHH+A/R组细胞的存活率升高明显,与DHL+A/R组比较有显著差异(P<0.05)。结论丹红注射液预处理可减轻心肌细胞A/R损伤,其机制可能为丹红注射液增加了细胞的内源性抗氧化剂活性,改善了心肌细胞能量代谢;且在一定范围内呈剂量依赖性,随着剂量的升高,保护作用更明显。

Objective To explore the protective effects of Danhong injection （DH） on neonatal rat myocardial cells with A/R injury and the relative mechanisms. Methods established neonatal rat myocardial cells anoxia/reoxygenation （A/R） injury model The myocardial cells were isolated from SD neonatal rats and cultured,followed by random division into five groups after being cul- tured for 3d （the experiment was duplicated 6 times in each groups）：normal control group,A/R group,DH Low dose preconditioning group（DHL＋A/R group）,DH moderate dose preconditioning group（DHM＋A/R group）,DH high dose preconditioning group（ DHH＋A/ R group）.Various teclmiques were adopted to detect the cells and cellular products： the levels of myocardial cells viability rate,the levels of lactate dehydrogenase （LDH）and superoxide dismutase（SOD） of culture fluid,the levels of intraeellular adenosine triphos- phate（ATP）. Results Compared with the control group,the myocardial cells viability and the levels of SOD and ATP of MR group significantly decreased（P〈0.01,respectively）,whereas LDH significantly increased （P〈0.01, respectively）. Compared with A/R group, the myocardial cells viability and the levels of SOD and ATP of each DH＋A/R group was significantly increased （P〈0.01, respec- tively） ,while released LDH significantly decreased （P〈0.01, respectively）. Conclusion DH could protect myocardial cells against A/R injury ,the mechanisms of which may be that Danhong injection increased cell endogenous antioxidant activity and improved the myocardial cell energy metabolism.In a certain range this function was dose dependent. As the dose increases, we can see more obvious protective effect.