Study on regulation of human hypertrophic scar fibroblast behavior by miR-211-5p via the TGF-β/Smad2 signaling pathway
Objective To investigate the mechanisms of microRNA-211-5p(miR-211-5p)in regulation of proliferation,apoptosis,migration,invasion,and collagen synthesis of human hyper-trophic scar fibroblasts(HSFBs)via the transforming growth factor(TGF)-β1/Smad homolog 2(Smad2)signaling pathway.Methods HSFBs were randomly divided into control,miR-NC,miR-211-5p mimic,anti-miR-21 1-5p,and miR-211-5p mimic+SB431542(TGF-β1/Smad2 signaling pathway inhibitor)groups.After 72 hours of continuous culture,quantitative real-time reverse tran-scription polymerase chain reaction(qRT-PCR)was used to detect miR-211-5p expression,western blot was employed to assess TGF-β1 and Smad2 protein levels,methyl-thiazoldiphenyl-tetrazolium(MTT)assay was performed to measure cell proliferation,flow cytometry was utilized to analyze ap-optosis rates,Transwell chambers were applied to evaluate cell invasion and migration,and western blot was again utilized to quantify type Ⅰ collagen(Col-Ⅰ)and type Ⅲ collagen(Col-Ⅲ)protein ex-pressions.An animal model of hypertrophic scars was established in rats using a constant temperature and pressure electric scalding apparatus.Following successful modeling,rats in each group received tail vein injections of miR-NC or miR-211-5p mimic.Scar healing was assessed,and histopathological changes in scar tissue were observed via hematoxylin and eosin(HE)staining.Results Compared with the control and miR-NC groups,the miR-211-5p mimic group exhibited increased miR-21 1-5p,TGF-β1,and Smad2 protein expressions,enhanced cell proliferation,reduced invasion and migra-tion capabilities,decreased apoptosis rates,and elevated Col-Ⅰ and Col-Ⅲ protein expressions(P＜0.05).Conversely,the anti-miR-211-5p group displayed opposite trends,and significant differences were observed between the anti-miR-211-5p and miR-211-5p mimic groups for the afore-mentioned indicators(P＜0.05).Compared to the miR-NC group,the miR-211-5p mimic group had higher scar healing rates and fibroblast counts(P＜0.05).Compared to the miR-21 1-5p mimic group,the miR-21 1-5p mimic+SB431542 group showed reduced TGF-β1,Smad2 protein expres-sions,and weakened cell invasion and migration capabilities(P＜0.05).Conclusion MiR-211-5p may regulate proliferation,apoptosis,migration,invasion,and collagen synthesis of HSFBs by activating the TGF-β1/Smad2 signaling pathway.
万方数据
维普
