中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (4): 603-608.doi: 10.3969/j.issn.2095-4344.2017.04.019

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

十子代平方影响小鼠MIN6细胞凋亡及MEK1/2、ERK1/2的表达

程瑞婷,董玉山,李继安,喇孝瑾,田春雨,高秀娟,周雪梅,付茜茹,吴  博,董思圻   

  1. 华北理工大学中医学院,河北省唐山市  063000
  • 收稿日期:2016-12-12 出版日期:2017-02-08 发布日期:2017-03-13
  • 通讯作者: 通讯作者:喇孝瑾,硕士,实验师,华北理工大学中医学院,河北省唐山市 063000
  • 作者简介:程瑞婷,女,1989年,河北省赤城县人,汉族,2016年华北理工大学毕业,硕士,医师,主要从事中医糖脂代谢疾病研究。
  • 基金资助:

    河北省国际科技合作计划项目(13392502D);河北省中医药管理局科研计划项目(2013066)

Effects of Shizidaiping formula on MIN6 cell apoptosis and expressions of MEK1/2 and ERK1/2

Cheng Rui-ting, Dong Yu-shan, Li Ji-an, La Xiao-jin, Tian Chun-yu, Gao Xiu-juan, Zhou Xue-mei, Fu Qian-ru,Wu Bo, Dong Si-qi   

  1. Institute of Traditional Chinese Medicine of North China University of Science and Technology, Tangshan 063000, Hebei Province, China
  • Received:2016-12-12 Online:2017-02-08 Published:2017-03-13
  • Contact: Corresponding author: La Xiao-jin, Master, Experimentalist, Institute of Traditional Chinese Medicine of North China University of Science and Technology, Tangshan 063000, Hebei Province, China
  • About author:Cheng Rui-ting, Master, Physician, Institute of Traditional Chinese Medicine of North China University of Science and Technology, Tangshan 063000, Hebei Province, China
  • Supported by:

    the International Cooperation Science and Technology Program of Hebei Province, No.13392502D; the Scientific Research Program of Administration of Traditional Chinese Medicine of Hebei Province, No. 2013066

摘要:

文章快速阅读:

文题释义:
十子代平方:由菟丝子、女贞子、莱菔子、苏子、车前子、茺蔚子、枸杞子、决明子、白芥子、五味子10味临床常用中药组成,研究表明该方可以有效抑制3T3-L1 脂肪细胞增殖,改善其胰岛素抵抗。
MIN6细胞:MIN6小鼠胰岛β细胞株是肉瘤病毒40(SV40)通过T细胞转染非肥胖糖尿病小鼠(NOD鼠)得到的胰岛细胞瘤株,在高糖浓度时胰岛素的分泌明显增加,其胰岛素分泌量和S型反应曲线类似于培养的正常胰岛细胞 。
ERK1/2:细胞外调节蛋白激酶(extracellular regulated protein kinases, ERK)分为ERK1 和ERK2,统称为ERK1 /2,主要被各种生长因子、离子射线、过氧化氢等磷酸化而激活,进入细胞核作用于E1k-1,c-myc,c-fos,c-jun,ATF,NF-kB和AP-1等转录因子,促进某些基因的转录与表达,和细胞的增殖与分化密切相关。
摘要
背景:
胰岛细胞渐进凋亡与长期高糖高脂损伤密切相关。
目的:观察十子代平方对高糖高脂损伤的MIN6细胞凋亡和胰岛素分泌的作用,分析十子代平方对MIN6细胞的保护作用和凋亡相关机制。
方法:细胞实验以MIN6细胞株作为实验对象,设正常组、模型组、二甲双胍组、十子代平方-高、十子代平方-中、十子代平方-低。CCK-8试剂盒检测十子代平方对MIN6细胞活性的影响;ELISA法检测其对胰岛素分泌的影响;Annexin V-FITC&PI 双染法检测其对凋亡的影响,Western Blotting法检测MEK1/2、ERK1/2和p-ERK1/2的表达水平。
结果与结论:①十子代平方能显著改善MIN6细胞在高糖高脂损伤下的细胞活力(P < 0.05),减少细胞早期凋亡,增加高糖刺激后上清液中的胰岛素水平(P < 0.05),提高MEK1/2、ERK1/2和p-ERK1/2表达水平(P < 0.05);②结果说明,十子代平方能够提高高糖高脂损伤的胰岛MIN6细胞活力、减轻胰岛素分泌负荷、抑制MIN6细胞凋亡,从而保护MIN6细胞。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0002-8577-9070(程瑞婷)

关键词: 组织构建, 组织工程, 十子代平方, MIN6, 细胞活性, 凋亡, 胰岛素, MEK1/2, ERK1/2

Abstract:

Abstract
BACKGROUND
: Apoptosis of islet cells is closely related to the long-term hyperglycemia- and hyperlipemia-induced injuries.
OBJECTIVE: To observe the effect of Shizidaiping formula on the apoptosis and insulin secretion in MIN6 cells under the high glucose and lipid environment, and to explore the protective effect of Shizidaiping formula and the related apoptosis mechanism.
METHODS: MIN6 cells were divided into normal, model, melbine, low-, medium-and high-dose Shizidaiping formula groups. The cell activity was examined by cell counting kit-8, the insulin secretion was measured by ELISA, the rate of apoptosis was measured by Annexin V-FITC&PI and the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 were examined by western blot assay.
RESULTS AND CONCLUSION: Shizidaiping formula significantly improved MIN6 cell activity under high glucose and lipid condition (P < 0.05), decreased early cell apoptosis, increased the level of insulin stimulated by low glucose in cell supernatant (P < 0.05), and improved the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 (P < 0.05). These results suggest that Shizidaiping formula can protect islet cells from hyperglycemia and hyperlipemia damage by improving the activity of MIN6 cells, reducing the insulin secretion and inhibiting the apoptosis of pancreatic β cells in MIN6 cells.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Drugs, Chinese Herbal, Islets of Langerhans, Apoptosis, Tissue

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