苓桂术甘汤含药血清对过氧化氢诱导的乳鼠原代心肌细胞氧化应激损伤及细胞凋亡的影响
Effect of Serum Containing Linggui Zhugan Decoction on Hydrogen Peroxide- induced Oxidative Stress Injury and Apoptosis of Primary Cardiomyocytes from Neonatal Mice
  
DOI:
中文关键词:  苓桂术甘汤  含药血清  过氧化氢  心肌细胞  氧化应激  细胞凋亡
英文关键词:Linggui Zhugan Decoction  Drug- containing serum  Hydrogen peroxide  Cardiomyocyte  Oxidative stress  Cell apoptosis
基金项目:国家自然科学基金项目(81202631,81373533);安徽高校自然科学研究重点项目(KJ2017A303);安徽中医药大学研究生科技创新基金一般项目(2017YB12)
作者单位
丁婉雪,葛瑞瑞,黄金玲,周 鹏,王 靓,甘贤兵,施 慧,范晓芸 1.安徽中医药大学研究生院安徽 合肥 2300122.安徽中医药大学中西医结合学院安徽 合肥 2300123.中药复方安徽省重点实验室安徽 合肥 2300124.安徽中医药大学护理学院安徽 合肥 230012 
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中文摘要:
      目的 观察苓桂术甘汤含药血清对H2O2诱导的心肌细胞氧化应激损伤及细胞凋亡的影响。方法 建立H2O2诱导的心肌细胞氧化应激损伤模型,将心肌细胞随机分为正常对照组、H2O2模型组、空白大鼠血清(20%)组、苓桂术甘汤含药血清(5%、10%、20%)组,用苓桂术甘汤含药血清(5%、10%、20%),分别预处理心肌细胞12 h后,加入100 μmol/L H2O2继续培养6 h,用MTT法测定心肌细胞存活率,比色法检测乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化酶(glutathione peroxidase ,GSH- Px)活性及丙二醛(malonic dialdehyde,MDA)水平,用荧光核染色法观察细胞凋亡形态学变化,流式细胞术(Annexin Ⅴ- FITC/PI)检测细胞凋亡率,荧光探针法测定氧自由基水平。结果 与正常组比较,模型组心肌细胞活力明显降低(P<0.05),氧自由基、LDH活性及MDA水平均显著升高(P<0.05),SOD及GSH- Px活性均显著降低(P<0.05),细胞核浓染数目增加,细胞凋亡率明显升高(P<0.05);与模型组比较,苓桂术甘汤含药血清组心肌细胞活力明显升高(P<0.05),GSH- Px、SOD活性均显著升高(P<0.05),LDH活性,氧自由基、MDA水平均显著降低(P<0.05),心肌细胞核浓染减少,细胞凋亡率明显降低(P<0.05)。结论 苓桂术甘汤能够清除细胞内氧自由基,抑制心肌细胞凋亡,增强细胞活性,对心肌细胞氧化应激损伤具有保护作用。
英文摘要:
      Objective To investigate the effect of serum containing Linggui Zhugan Decoction on hydrogen peroxide (H2O2)- induced oxidative stress injury and apoptosis of primary cardiomyocytes. Methods Cardiomyocytes were randomly divided into normal group, H2O2 model group, blank rat serum (20%) group, and serum containing Linggui Zhugan Decoction groups (5%, 10%, and 20%). All cardiomyocytes except those in the normal group were used to establish a model of H2O2- induced oxidative stress injury, and the cardiomyocytes in the serum containing Linggui Zhugan Decoction groups were pretreated with serum containing Linggui Zhugan Decoction (5%, 10%, and 20%) for 12 hours and then cultured with 100 μmol/L H2O2 for 6 hours. MTT assay was used to measure the survival rate of cardiomyocytes; colorimetry was used to measure the activities of lactate dehydrogenase (LDH), superoxide dismutase (SOD), and glutathione peroxidase (GSH- Px) and the level of malondialdehyde (MDA); fluorescence nuclear staining was used to observe the apoptosis and morphological changes of cardiomyocytes; flow cytometry (Annexin Ⅴ- FITC/PI) was used to measure the apoptosis rate of cardiomyocytes; the fluorescent probe method was used to measure the level of oxygen free radicals. Results Compared with the normal group, the model group had a significant reduction in cardiomyocyte viability (P<0.05), significant increases in oxygen free radicals, LDH activity, and MDA level (P<0.05), significant reductions in the activities of SOD and GSH- Px (P<0.05), and significant increases in the number of densely stained nuclei and the apoptosis rate of cardiomyocytes (P<0.05). Compared with the model group, the serum containing Linggui Zhugan Decoction groups had a significant increase in cardiomyocyte viability (P<0.05), significant increases in the activities of SOD and GSH- Px (P<0.05), significant reductions in LDH activity, oxygen free radicals, and MDA level (P<0.05), and significant reductions in the number of densely stained nuclei and the apoptosis rate of cardiomyocytes (P<0.05). Conclusion Linggui Zhugan Decoction can clear oxygen free radicals in cardiomyocytes, inhibit cardiomyocyte apoptosis, enhance cardiomyocyte activity, and protect cardiomyocytes against oxidative stress injury.
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