盐酸二甲双胍通过抑制PPARγ表达拮抗成骨细胞的糖毒性损害  

Metformin Hydrochloride Antagonizes Glucotoxicity in Osteoblasts by Restraining the Expression of PPARγ

作　　者：严孙杰[1] 李毅敏[2] 颜晓芳[1] 沈喜妹[1] 

机构地区：[1]福建医科大学附属第一医院内分泌科,福州350005 [2]福建省漳州市医院,福建漳州363000

出　　处：《中国药学杂志》2014年第2期121-125,共5页Chinese Pharmaceutical Journal

基　　金：福建省自然科学基金资助项目(2013J01295);福建省卫生厅青年课题基金(2010-2-26);福建医科大学教授学术发展基金(JS11015)

摘　　要：摘要：目的探讨盐酸二甲双胍（MF）在高糖环境中对成骨细胞的保护作用与过氧化物酶增殖激活物受体（PPAR）γ表达的关系。方法体外培养小鼠颅骨原代成骨细胞，分为5．5mmol·L11正常糖组、25mmol·L-1高糖＋不同浓度盐酸二甲双胍干预组（0、25、50、100μmol·L-1盐酸二甲双胍）；干预时间1周、2周、3周，半定量RT—PCR法检测细胞PPARγ、骨形成蛋白（BMP）一2的mRNA表达，Western印迹检测PPARγ的蛋白表达情况。结果与正常糖组相比，在相同干预时间高糖组细胞的PPARγ mRNA表达量增加（均P〈0．05），高糖干预3周细胞PPARγ蛋白表达量也明显增加（P〈0．05）；高糖组BMP-2mRNA表达量降低（均P〈0．05）。同一干预时间，高糖环境随盐酸二甲双胍浓度在0～100p．mol·L。内增加，细胞PPARγ／mRNA表达递减、BMP-2mRNA表达则递增（均P〈0．05）；随着干预时间从1周～3周延长，各高糖干预组PPARγ mRNA表达量则逐渐上升，BMP-2mRNA表达量变化则相反（均P〈0．05）。干预3周后，随着盐酸二甲双胍浓度在0～100μmol·L-1内增高，高糖环境PPAR7蛋白表达量逐渐减少（P〈0．05）。结论病理浓度的葡萄糖导致成骨细胞分化不良，成骨能力下降可能与诱导PPARγ mRNA及蛋白表达增高有关：应用盐酸二甲双胍可抑制PPARγ的表达，保护成骨细胞。OBJECTIVE To investigate the protective effects of metformin hydrochloride （MF） on osteoblasts which were ex- posed to constant high glucose condition, and the relationship between MF and the expression of peroxisome proliferator-activated recep- tor （PPAR） 7. METHODS Mouse cranium osteoblasts were cultured in vitro and divided into 5.5 mmol· L-1 normal glucose ＋ 0 μmol· L-1 MF group, 25 mmol · L-1 high glucose with different concentration of MF （0, 25, 50, 100μmol· L-1 MF, respectively） and intervened for 1,2 and 3 w, then the mRNA expression levels of PPARγ and bone morphogenetic protein （BMP） -2 were detected by RT-PCR, and the protein expression levels of PPARγ was examined by Western blot. RESULTS （1）The mRNA expression levels of PPARγwas marked higher in high glucose group compared to the control group with the same intervene time （ all P 〈 0. 05 ） ; when the intervention time proloilged to 3w, the protein expression of PPARγ increased significantly （P 〈 0. 05 ） ; the mRNA expression lev- els of BMP-2 in the high glucose group was lower than in the control group （ all P 〈 0. 05）. （3）At the same intervening time, the ex- pression of PPAR7 mRNA decreased while BMP-2 mRNA increased （ all P 〈 0. 05 ） with the increasing concentration of MF （ from 0 μmol · L-1 to 100 μ mol· L-1 ） ; when the intervention time prolonged from lw to 3w, the mRNA expression levels of PPARγ in- creased in all the high glucose groups while the mRNA expression levels of BMP-2 decreased （ all P 〈 0. 05）. （3）When interviewed for 3w, the protein expression of PPARγ decreased gradually with the increasing concentration of MF in high glucose condition （P 〈 0. 05）. CONCLUSION Pathological concentration of glucose could lead to poorly differentiated of osteoblasts, which might associa- tion with the enhanced mRNA and protein expression levels of PPARγ; MF can protect osteoblasts by restraining the expression of PPARγ.

关 键 词：盐酸二甲双胍 成骨细胞 过氧化物酶增殖激活物受体γ 骨形成蛋白2 

分 类 号：R692.5[医药卫生—泌尿科学]