意大利牛舌草乙醇提取物抗缺氧/复氧心肌细胞氧化应激损伤的研究

The study on the protect effect of the ethanol extracts fromAnchusaitalica Retz on hypoxia reoxygenationinduced oxidative stress injury in primary cardiomyocytes

目的研究意大利牛舌草(Anchusaitalica Retz)乙醇提取物的预处理对缺氧/复氧心肌细胞氧化应激损伤的影响。方法建立缺氧/复氧体外小鼠原代心肌细胞模型以模拟缺血再灌注损伤。实验分为6组:正常对照组、缺氧/复氧组、阳性对照组(缺氧/复氧+1.25 mg/mL葛根素)、低剂量乙醇提取物处理组(缺氧/复氧+1.25mg/mL意大利牛舌草乙醇提取物)、中剂量乙醇提取物处理组(缺氧/复氧+2.50 mg/mL意大利牛舌草乙醇提取物)、高剂量乙醇提取物处理组(缺氧/复氧+5.00 mg/mL意大利牛舌草乙醇提取物)。采用MTT法检测原代心肌细胞存活率,采用酶标法检测细胞培养上清液中乳酸脱氢酶(LDH)水平、细胞内的丙二醛(MDA)含量和细胞内超氧化物歧化酶(SOD)的活力。结果与正常对照组比较,缺氧/复氧组呈现时间依赖性地降低原代心肌细胞存活率,差异有统计学意义(P<0.01);与缺氧/复氧组比较,乙醇提取物处理组、阳性对照组LDH、MDA含量降低,SOD活力提高,差异有统计学意义(P<0.01或P<0.05)。结论意大利牛舌草乙醇提取物可能通过干预平衡氧化应激产物的生成和抗氧化应激的酶系统来减少缺氧/复氧所致原代心肌细胞氧化应激损伤。

Objective To investigate the effect of the ethanol extracts from Anchusaitalica Retz on hypoxia/ reoxygenation induced oxidative stress injury in primary cardiomyocytes. Methods To simulate ischemia- reperfusion injury, an anoxia/reoxygenation model is set up by cultivating the original generation of myo-cardial cells in mice in vitro. There were six experimental groups, Normal control group, hypoxia/reoxy- genation （H/R） group, positive control group （H/R＋ 1.25 mg/mL puerarin）, low dose ethanol extracts group （H/R＋ 1.25 mg/mLethanol extracts）, middle dose ethanol extracts group （H/R＋ 2.5 mg/mL etha- nol extracts）, high dose ethanol extracts group （H/R＋5.00 mg/mL ethanol extracts）. Survival rate of primary cardiomyocytes were determined by MTT method. The levels of lactate dehydrogenase （LDH）,intracellular superoxide dismutase （SOD） and malondialdehyde （MDA） were assessed by microplate reader. Results Compared with control group, survival rate of hypoxia/reoxygenation （H/R） group cells reduced, and the decline was a time-dependence effect. Compared with H/R group, ethanol extracts and puerarin could reduce the content of LDH and MDA and improve intracellular SOD activity, and the difference was statistically significant （P〈0.01 or P〈0.05）. Conclusion Ethanol extracts from Anchusaitalica Retz could reduce oxidative stress injury induced by H/R. The mechanism might be connected with balancing the oxidativestress products and anti-oxidative stress-related enzyme systems.