二甲双胍对小鼠颅骨成骨细胞糖毒性损害的保护作用  

Protective effect of metformin on glucotoxicity in mouse cranium osteoblasts

作　　者：严孙杰[1] 李毅敏[1] 沈喜妹[1] 颜晓芳[1] 

机构地区：[1]福建医科大学附属第一医院内分泌科,福建福州350005

出　　处：《中国新药与临床杂志》2011年第11期852-856,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：福建省自然科学基金资助项目(2009J01135);福建省卫生厅青年课题基金项目(2010-2-26)

摘　　要：目的探讨二甲双胍对高糖介导的成骨细胞增殖受限、凋亡及功能蛋白表达损害的干预作用。方法体外培养原代小鼠颅骨成骨细胞,分为正常糖浓度(5.5 mmol·L^(-1)葡萄糖)组,高糖(25 mmol·L^(-1)葡萄糖)组,二甲双胍低、中、高浓度(25 mmol·L^(-1)葡萄糖+25、50、100μmol·L^(-1)二甲双胍)组,分别干预48、72 h,MTT法测定细胞增殖活性,流式细胞仪检测细胞凋亡率。各实验组细胞加入成骨诱导培养基,干预时间延长至1、2、3 wk,比色法和放射免疫法检测碱性磷酸酶(ALP)、骨钙素的分泌水平。结果在相同干预时限,与正常糖浓度组比较,高糖组细胞增殖能力降低、早期凋亡率增加(均P<0.05);二甲双胍低、中、高浓度组,随二甲双胍浓度增高,细胞增殖能力呈上升趋势(P<0.05),而细胞早期凋亡率呈下降趋势(P<0.05)。干预时间由48 h延长至72 h,各组细胞增殖能力和早期凋亡率呈不同程度增加。干预时间延长至1、2、3 wk;正常糖浓度组细胞ALP和骨钙素的分泌水平呈增加趋势(P<0.05),而高糖组则呈下降趋势(P<0.05)。在相同干预时限,二甲双胍干预组细胞ALP和骨钙素分泌水平随二甲双胍浓度增高而增加(P<0.05)。结论二甲双胍能够拮抗成骨细胞的糖毒性损害,保护成骨细胞的成骨能力,且在一定的浓度范围内呈剂量依赖性。AIM To investigate the effects of metformin on the high glucose-induced impairment on proliferation, apoptosis and expression of functional protein in osteoblasts. METHODS Mouse cranium osteo- blasts were cultured in vitro and divided into normal glucose group （5.5 mmol .L^-1） , high glucose group （25 mmol-L^-1）, and low, middle, high metformin group （25 mmol .L^-1 glucose ＋ 25, 50 or 100 μmol-L^-1 metformin, respectively）. The cells were intervened for 48 and 72 hours. The cell proliferation ability was determined by MTT assay, and the cell apoptosis rate was detected by flow cytometry. Culture medium for bone formation was added into each group, and the intervention time was extended to 1, 2 and 3 weeks. Chromatometry and RIA were used to determine the secretion levels of alkaline phosphatase （ALP） and osteocalcin （OCN）, respectively. RESULTS At the same intervention time, compared with the normal glucose group, the high glucose group showed lower cell proliferation rate and higher early apoptosis rate （P 〈 0.05）. With the increasing concentration of metformin, the proliferation rate of osteoblasts was increased （P 〈 0.05）, and the apoptosis rate was decreased （P 〈 0.05）. Prolonged the intervention time from 48 to 72 hours, the proliferation and early apoptosis rate of cells were increased in all groups. When the intervention time was extended to 1,2 and 3 weeks, the secretion levels of ALP and OCN were increased in the normal glucose group （P 〈 0.05）, while decreased in the high glucose group. At the same intervention time, the secretion levels of ALP and OCN were increased with the increasing concentration of metformin （P 〈 0.05）. CONCLUSION Metformin could protect against high glucose-induced cytotoxicity and improve the bone-formation capacity of osteoblasts in a dose-dependent manner.

关 键 词：二甲双胍 成骨细胞 细胞凋亡 碱性磷酸酶 骨钙素 

分 类 号：R977.15[医药卫生—药品]