二甲双胍对糖基化终末产物诱导的成骨细胞氧化应激和凋亡的影响

中国临床药理学与治疗学 ›› 2013, Vol. 18 ›› Issue (7): 743-748.

二甲双胍对糖基化终末产物诱导的成骨细胞氧化应激和凋亡的影响

甄东户, 刘丽娟, 成建国, 汤旭磊

兰州大学第一医院内分泌科,兰州 730000,甘肃

收稿日期:2012-10-10 修回日期:2013-04-08 出版日期:2013-07-26 发布日期:2013-06-20
通讯作者: 汤旭磊,男,博士,教授,硕士生导师,主任医师,主要从事骨质疏松以及抗氧化机制研究。Tel: 0931-8356860 E-mail: xulei_tang@126.com
作者简介:甄东户,女,博士,副主任医师,主要从事糖尿病性骨质疏松研究。Tel: 0931-8356894 E-mail: zhdh8279@163.com
基金资助:
兰州大学中央高校基本科研业务费专项资金资助自由探索项目(lzujbky-2010-199);兰州大学第一医院院级科研业务费(2010)

Effects of metformin on intracellular reactive oxygen species and apoptosis induced by advanced glycation end products on rat cranioaural osteoblasts

ZHEN Dong-hu, LIU Li-juan, CHEN Jian-guo, TANG Xu-lei

Department of Endocrinology, the First Hospital of Lanzhou University, Lanzhou 730000, Gansu, China

Received:2012-10-10 Revised:2013-04-08 Online:2013-07-26 Published:2013-06-20

摘要/Abstract

摘要： 目的: 观察二甲双胍对糖基化终末产物(AGEs)诱导的大鼠颅骨成骨细胞内氧化应激产物活性氧簇(ROS)、细胞早期凋亡的影响。方法: 分离培养大鼠颅骨成骨细胞,2′7′-二乙酰二氯荧光素(DCFH-DA)作为荧光探针,流式细胞检测技术检测细胞内ROS水平,膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/ PI)双染色分析细胞早期凋亡率。结果: 与未经葡萄糖处理的牛血清白蛋白(BSA)组对比,500 μg/mL AGEs显著促进成骨细胞内ROS形成和细胞凋亡(均P＜0.01);给予二甲双胍 (100~500 μmol/L)呈浓度依赖性抑制BSA组及AGEs组成骨细胞内ROS形成和细胞凋亡,浓度分别为500、400 μmol/L 时,对细胞内ROS形成和细胞凋亡的抑制作用达到最强。结论: AGEs显著诱导原代成骨细胞内ROS的形成和细胞凋亡,而二甲双胍能够呈浓度依赖性抑制AGEs诱导的成骨细胞内ROS的形成和细胞凋亡,减轻AGEs对成骨细胞的损害。

关键词: 二甲双胍, 糖基化终末产物, 活性氧簇, 凋亡

Abstract: AIM: To explore the effects of metformin (MF) on intracellular reactive oxygen species (ROS) and apoptosis in advanced glycation end products (AGEs)-induced rat cranioaural osteoblasts.METHODS: Rat cranioaural osteoblasts were isolated and cultured. The fluorescence of 2′, 7′-dichlorofluorescin (DCF) was measured by flow cytometry as a mean of estimating the formation of ROS, the cell apoptosis was detected by double-staining (Annexin V-FITC/ PI).RESULTS: With 500 μg/mL AGEs, intracellular ROS and the cell apoptosis were markedly increased(both P＜0.01), as compared with that observed with bovine serum albumin(BSA) group; MF(100-500 μmol/L) decreased intracellular ROS in both BSA group and AGEs group in a dose-dependent manner, with maximal inhibition attained at a concentration of 500 μmol/L. Introducing different concentrations of metformin (100-500 μmol/L) into the medium significantly lowered the number of apoptotic cells in a dose-dependent manner in both BSA group and AGEs group and maximal inhibition was reached at a concentration of 400 μmol/L.CONCLUSION: AGEs markedly induced intracellular ROS and the cell apoptosis. However, metformin significantly decreased intracellular ROS and apoptosis, which could ameliorate deleterious effects of AGEs on osteoblasts.

Key words: Metformin, Glycation end products, Reactive oxygen species, Apoptosis

中图分类号:

R965.2

甄东户, 刘丽娟, 成建国, 汤旭磊. 二甲双胍对糖基化终末产物诱导的成骨细胞氧化应激和凋亡的影响[J]. 中国临床药理学与治疗学, 2013, 18(7): 743-748.

ZHEN Dong-hu, LIU Li-juan, CHEN Jian-guo, TANG Xu-lei. Effects of metformin on intracellular reactive oxygen species and apoptosis induced by advanced glycation end products on rat cranioaural osteoblasts[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(7): 743-748.

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