中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (28): 5241-5244.doi: 10.3969/j.issn.1673-8225.2011.28.028

• 组织构建与中医药 tissue construction and traditional Chinese medicine • 上一篇    下一篇

人肝癌细胞胰岛素抵抗模型建立及有效中药成分的筛选

刘志霞，韩淑英，李继安   

  1. 河北联合大学中医学院，河北省唐山市  063000
  • 收稿日期:2011-02-06 修回日期:2011-05-17 出版日期:2011-07-09 发布日期:2011-07-09
  • 通讯作者: 李继安，博士，主任医师，教授，河北联合大学中医学院，河北省唐山市 063000 lnyy@vip.sina.com
  • 作者简介:刘志霞★，女，1977年生，汉族，河北省唐山市人，2011年河北联合大学毕业，硕士，医师，主要从事中医治疗2型糖尿病的相关研究。
  • 基金资助:

    课题受中国科技部国际科技合作项目(2008DFA31050)与河北省科技厅国际科技合作项目(08396423D)资助。

Establishment of the insulin resistance model with HepG2 cells and screening of active ingredients of traditional Chinese medicine

Liu Zhi-xia, Han Shu-ying, Li Ji-an   

  1. College of Traditional Chinese Medicine of Hebei United University, Tangshan  063000, Hebei Province, China
  • Received:2011-02-06 Revised:2011-05-17 Online:2011-07-09 Published:2011-07-09
  • Contact: Li Ji-an, Doctor, Chief physician, Professor, College of Traditional Chinese Medicine of Hebei United University, Tangshan 063000, Hebei Province, China lnyy@vip.sina.com
  • About author:Liu Zhi-xia★, Master, Physician, College of Traditional Chinese Medicine of Hebei United University, Tangshan 063000, Hebei Province, China
  • Supported by:

    International Cooperation Program of Ministry of Science and Technology, No. 2008DFA3150*; International Cooperation Program of Hebei Provincial Science and Technology Department, No. 08396423D*

PDF

572

被引次数

51

摘要：

背景：目前，复方中药、单味中药在体内降糖作用及其降糖机制研究较多，但体外尤其是中药单体成分对胰岛素抵抗细胞有何影响尚不清楚。
目的：体外建立人肝癌细胞(HepG2)胰岛素抵抗模型，并初步筛选可有效改善胰岛素抵抗的中药有效成分。
方法：用不同浓度的胰岛素对HepG2细胞进行不同时间的诱导，通过MTT法对细胞活性评价及葡萄糖氧化酶法对HepG2细胞葡萄糖消耗量测定，明确建立稳定的HepG2胰岛素抵抗模型的胰岛素诱导浓度及诱导时间。模型建立后，应用不同浓度的齐墩果酸、药根碱、阿魏酸、大黄酸、马钱苷、葛根素、大豆苷分别作用于胰岛素抵抗细胞24 h，用葡萄糖氧化酶法分别观察不同浓度的上述中药成分对胰岛素抵抗模型HepG2细胞葡萄糖消耗的影响，MTT法对各组细胞活性进行评价。
结果与结论：HepG2细胞在10-6  mol/L浓度的胰岛素中作用24 h，葡萄糖消耗量明显减少(P < 0.01)，说明实验成功诱导出稳定人肝癌细胞胰岛素抵抗模型。10-5 mol/L浓度胰岛素组的胰岛素抵抗更明显(P < 0.01)。各时间点10-5 mol/L浓度胰岛素作用的细胞成活率逐渐降低，死亡细胞增多(P < 0.05)。齐墩果酸、药根碱、阿魏酸、大黄酸、马钱苷、葛根素、大豆苷均有改善细胞胰岛素抵抗的作用。其中，质量浓度2×10-1 g/L药根碱、大黄酸、葛根素和齐墩果酸，2×10-5 g/L马钱苷和阿魏酸对改善人肝癌细胞胰岛素抵抗效果较好(P < 0.01)。

关键词: 胰岛素抵抗, HepG2细胞, 葡萄糖消耗, 细胞模型, 中医药, 组织构建

Abstract:

BACKGROUND: At present, there are many studies on the hypoglycemic effect and its mechanism of Chinese medicinal compounds or single herbs in vivo, but the effect of the traditional Chinese medicinal monomers on insulin resistance cells in vitro has been rarely reported.
OBJECTIVE: To establish human hepatoma carcinoma cell (HepG2 cell) model of insulin resistance in vitro and to initially select the effective ingredients in the traditional Chinese medicine for improving insulin resistance.
METHODS: HepG2 cells were induced by insulin of different concentrations for different time. Through the estimation of the cell activity with MTT method and glucose consumption with glucose oxidase method, the concentrations and induce time of insulin that can induce the stable insulin resistance cell model were ascertained. After model establishment, the model cells were treated with oleanolic acid, jatrorrhizine, ferulic acid, rhein, loganin, puerarin and daidzin of different concentrations for 24 hours, and the influences on the model cells regarding glucose consumption were observed through the glucose oxidase method and the cytoactive were evaluated by the MTT method .
RESULTS AND CONCLUSION: In the group of 10-6 mol/L at the time of 24 hours, the glucose consumption was decreased obviously compared with the normal group (P < 0.01), showing that stable insulin resistance models were induced successfully. The insulin resistance in the group of 10-5 mol/L was more evident compared with that in the normal group (P < 0.01), but the cell survival rate was decreased gradually with time and the number of dead cells were increased gradually (P < 0.05). The oleanolic acid, jatrorrhizine, ferulic acid, rhein, loganin, puerarin and daidzin can improve the insulin resistance of cells. 2×10-1 g/L of jatrorrhizine, rhein, puerarin and oleanolic acid, as well as 2×10-5 g/L of loganin and ferulic acid have better effects on improving the insulin resistance (P < 0.01).

中图分类号: