Effects of metformin on proliferation andosteogenesis of FLSs in interfacial membrane through AMPK pathway
10.3969/j.issn.1006-5725.2018.05.019
- VernacularTitle:二甲双胍通过AMPK通道对界膜中滑膜成纤维细胞的增殖和成骨作用的研究
- Author:
Kun CHEN
1
;
Qi LI
Author Information
1. 南方医科大学珠江医院 广州 510280
- Keywords:
metformin;
fibroblast-like synoviocytes;
AMPK;
proliferation;
osteogenesis
- From:
The Journal of Practical Medicine
2018;34(5):769-773,777
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate whether metformin can inhibit the growth of synovial fibroblasts (FLSs)via loosening interfacial membrane and promoting osteogenic differentiation through AMPK channels. Methods The extraction of primary and FLSs of 4-5 passages after purification used for experiment were treated with Metformin and bone morphogenic agonists(ascorbicacid,betaglycerophosphateand dexamethasone treat-ment),respectively.The electron microscopy and immunocytochemical staining identified fibroblasts;MTT detect-ed FLSs growth inhibition rate,which treated with drugs in different concentration and time;Alkaline phosphatase (ALP)staining and alizarin red staining were used to detect the changes of ALP activity and osteogenesis with time after FLSs treated with medicine;Western Blot was used to detect AMPK and phosphorylation level after drug treat-ment.Results FLSs were with spindle shape,large cell body and small amount of polygonal cells under electron microscope. The immunocytochemical staining of Keratin and Vimentin was negative and positive,respectively. The inhibition rate of metformin on FLSs proliferation was increased in a concentration dependent manner. Strong positive results were found in ALP and alizarin red staining in the metformin group,which was similar with the bone morphogenetic agonist group. Conclusion Metformin can inhibit FLSs growth and promote the synthesis of alkaline phosphatase and deposition of extracellular calcium nodules through activation of AMPK channel in a time and concentration dependent manner.