STAT signaling pathway mediates high glucose induced cardiac fibroblasts proliferation and collagen deposition in vitro
10.3760/cma.j.issn.0253-3758.2015.05.016
- VernacularTitle:STAT信号通路介导高糖诱导的鼠心肌成纤维细胞增殖和胶原沉积
- Author:
Bin DAI
1
;
Mei ZHU
;
Wenling SU
;
Mingcai QIU
;
Hong ZHANG
Author Information
1. 300052,天津医科大学总医院内分泌科
- Keywords:
Myocardium;
Fibroblasts;
Cell proliferation;
Collagen;
Signal transducers and activator of transcriptions
- From:
Chinese Journal of Cardiology
2015;43(5):442-447
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the signal transducers and activator of transcriptions (STATs) protein expression changes and investigate the functional role of STATs pathway in case of high glucoseinduced cardiac fibroblasts (CFs) proliferation and collagen deposition in vitro.Methods Rat cardiac fibroblasts were isolated from 1-to 3-day-old SD rats,cells from the second to fourth passages were used for the experiment.CFs were cultured in Dulbecco's modified Eagle's medium,supplemented with 5.5 mmol/L glucose(NG),5.5 mmol/L glucose plus 19.4 mmol/L mannose(OC) or 25 mmol/L glucose(HG) in the presence of absence of STAT1 inhibitor (fludarabine,FLU) and STAT3 inhibitor (S3I-201).After 24 h and 48 h culture in vitro,the proliferation of CFs was measured by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl2H-tetrazolium bromide (MTT) assay.After 12 h and 24 h culture in vitro,the production of type Ⅰ and Ⅲ collagen was evaluated using real-time quantitative PCR and ELISA.After 0,30,60 and 120 min culture in vitro,the phosphorylated expression of STAT1 and STAT3 was analyzed by Western blot.Results CFs proliferation was significantly enhanced post 24 h and 48 h HG stimulation,and procollagen Ⅰ and Ⅲ mRNA expression was significantly upregulated post 12 h and 24 h HG stimulation.Deposition of collagen Ⅰ and Ⅲ was also significantly increased post 24 h and 72 h HG stimulation.STAT1 phosphorylation in CFs was increased after 120 min HG stimulation and STAT3 phosphorylation in CFs was increased post 60 min and 120 min HG stimulation.FLU and S3I-201 could inhibit HG-induced CFs proliferation and suppress of which was stimulated by FLU and S3I-201 could both suppress upregulated procollagen Ⅰ and Ⅲ mRNA expression and the deposition of collagen types Ⅰ and Ⅲ post HG stimulation.STAT1 phosphorylation inhibition resulted in less mRNA downregulation of procollagen type Ⅱ than that of procollagen type Ⅰ post 12 h HG stimulation.The STAT3 phosphorylation inhibition resulted in more significantly upregulated procollagen type Ⅲ mRNA expression than procollagen type Ⅰ mRNA expression at 12 h post HG stimulation.Conclusion HG could enhance the protein expression of phosphorylated STAT1 and STAT3 in CFs,which are responsible for HG-induced increased CFs proliferation and collagen deposition in vitro.
