吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (06): 1248-1255.doi: 10.13481/j.1671-587x.20190610

• 基础研究 • 上一篇    下一篇

人参提取物对棕榈酸诱导心肌细胞损伤的保护作用及其机制

娄婷婷1, 黄清霞2, 李香艳1, 赵大庆1   

  1. 1. 长春中医药大学吉林省人参科学研究院, 吉林 长春 130117;
    2. 长春中医药大学附属医院中医药研究中心, 吉林 长春 130021
  • 收稿日期:2019-02-14 出版日期:2019-12-05 发布日期:2019-12-05
  • 通讯作者: 赵大庆,研究员,博士研究生导师(Tel:0431-86177401,E-mail:zhaodaqing1963@163.com) E-mail:zhaodaqing1963@163.com
  • 作者简介:娄婷婷(1990-),女,吉林省长春市人,在读医学硕士,主要从事人参功效生物分子机制方面的研究。
  • 基金资助:
    国家科技部"十三五"重点研发计划项目资助课题(2017YFC1702103);吉林省科技厅中青年科技创新领军人才及团队项目资助课题(20190101010JH);吉林省长春市科技局医药健康产业科技创新重大专项资助课题(18YJ013)

Protective effect of ginseng extract on cardiomyocyte injury induced by palmitic acidand its mechanism

LOU Tingting1, HUANG Qingxia2, LI Xiangyan1, ZHAO Daqing1   

  1. 1. Jilin Provincial Ginseng Academy, Changchun University of Chinese Medicine, Changchun 130117, China;
    2. Research Center of Traditional Chinese Medicine, Affiliated Hospital, Changchun University of Traditional Chinese Medicine, Changchun 130021, China
  • Received:2019-02-14 Online:2019-12-05 Published:2019-12-05

摘要: 目的:观察人参提取物对棕榈酸(PA)引起的脂毒性心肌细胞损伤的保护作用,并探讨其相关作用机制。方法:体外培养H9c2细胞。油红O染色检测不同浓度(100、200和1000 μmol·L-1) PA作用后H9c2细胞中脂滴水平,MTT法检测不同浓度(200、400、600和800μmol·L-1) PA作用后H9c2细胞存活率,流式细胞术检测不同浓度(200、400、600和800μmol·L-1) PA作用后H9c2细胞凋亡率,筛选PA作用浓度和时间。依据筛选结果选择PA作用浓度为100和200 μmol·L-1,作用时间为24 h。将H9c2细胞随机分为对照组、PA组(100和200μmol·L-1)和不同浓度(0.2、2.0和20.0 mg·L-1)人参提取物组。流式细胞术和FITC-Annexin Ⅴ/PI双染法检测各组H9c2细胞凋亡率,JC-1染色法检测各组H9c2细胞中线粒体膜电位(MMP)水平,DCFH-DA染色法检测各组H9c2细胞中活性氧(ROS)水平。结果:与对照组比较,100、200和1 000 μmol·L-1PA组H9c2细胞中脂滴水平明显升高(P<0.01)。与对照组比较,200、400、600和800 mmol·L-1 PA组H9c2细胞存活率逐渐降低(P<0.01),细胞凋亡率逐渐升高(P<0.01)。不同浓度人参提取物作用24h后,与100和200μmol·L-1PA组比较,2.0和20.0 mg·L-1人参提取物组H9c2细胞中脂滴水平降低(P<0.01),细胞凋亡率逐渐降低(P<0.01),ROS水平逐渐降低(P<0.01);各浓度人参提取物组H9c2细胞中MMP水平逐渐降低(P<0.01)。结论:人参提取物可以抑制PA诱导的心肌细胞凋亡,其机制可能与受损H9c2细胞中ROS和MMP水平下调有关。

关键词: 人参提取物, 脂毒性, 细胞凋亡, 活性氧, 线粒体膜电位, 棕榈酸

Abstract: Objective: To investigate the protective effect of ginseng extract on the lipotoxic cardiomyocyte injury induced by palmitic acid (PA),and to clarify its possible mechanism. Methods: The H9c2 cells were cultured in vitro.The levels of lipid droplets in H9c2 cells after treated with different concentrations(100,200,and 1 000 μmol·L-1) of PA were detected by oil red O staining,the survival rates of H9c2 cells after treated with different concentrations(200,400,600,and 800 μmol·L-1) of PA were detected by MTT assay,and the apoptotic rates of H9c2 cells after treated with different concentrations(200,400,600,and 800 μmol·L-1) of PA were detected by flow cytometry.The concentration of PA and the action time were screened;according to the screening results,the PA concentrations were selected as 100 and 200 μmol·L-1,and the action time was 24 h.The H9c2 cells were randomly divided into control group,PA group (100 or 200 μmol·L-1) and and different concentrations (0.2,2.0,and 20.0 mg·L-1)of ginseng extract groups.Flow cytometry and FITC-Annexin Ⅴ/PI double staining were used to detect the apoptotic rates of the H9c2 cells in various groups,JC-1 staining was used to detect the levels of mitochondrial membrane potential (MMP) in the H9c2 cells in various groups,and DCFH-DA staining was used to detect the levels of intracellular reactive oxygen species (ROS) in the H9c2 cells in various groups. Results: Compared with control group,the levels of lipid droplets in the H9c2 cells in 100,200, and 1 000 μmol·L-1 PA groups were increased significantly (P<0.01).Compared with control group,the survival rates of the H9c2 cells in 200,400,600,and 800 mmol·L-1 PA groups were decreased (P<0.01),and the apoptotic rates of the H9c2 cells in 200,400,600,and 800 mmol·L-1 PA groups were increased(P<0.01).After treated with different concentrations of ginseng extract for 24 h,compared with 100 or 200 μmol·L-1 PA groups,the lipid levels of the H9c2 cells in 2.0 and 20.0 mg·L-1 ginseng extract groups were decreased(P<0.01),the apoptotic rates were decreased(P<0.01),and the levels of ROS were decreased (P<0.01); the levels of MMP in the H9c2 cells in different concentrations of ginseng extract groups were decreased (P<0.01). Conclusion: Ginseng extract can inhibit the apoptosis of the cardiomyocytes induced by PA,and its mechanism may be related to the down-regulation of ROS and MMP levels in the H9c2 cells.

Key words: ginseng extract, lipidtoxicity, apoptosis, reactive oxygen species, mitochondrial membrane potential, palmitic acid

中图分类号: 

  • Q27