Inhibitory effect of doxycycline on TGF-β1-induced human corneal stromal cell fibrosis
Objective:To investigate the inhibitory effect of doxycycline(Doxy)on Transforming Growth Factor-β1(TGF-β1)-induced human corneal stromal cell fibrosis.Methods:Human corneal stromal cells were extracted from corneal stromal lens after femtosecond laser-assisted small incision lenticule extraction(SMILE),and the corneal fibrosis model was established in vitro by inducing human corneal stromal cells with TGF-β1.Control group,TGF-β1 group and Doxy group were set up.The proliferation ability of each group was detected by cell counting kit-8(CCK-8)method,the proliferation ability was detected by scratch test,and the content of hydroxy-proline(HYP),a key factor in collagen synthesis,was determined by enzyme-related immunosorbent assay(ELI-SA).Reverse transcription-quantitative PCR(RT-qPCR)was used to detect the mRNA expression of the key fac-tors of fibrosis Thy-1 and Vimentin,and western blotting was used to detect the protein expression levels of Thy-1 and Vimentin.Results:Compared with the control group,TGF-β1 group could promote the proliferation and mi-gration of human corneal stromal cells,and increase the content of HYP,the expression of Thy-1 and Vimentin(all P＜0.05).Compared with the TGF-β1 group,the abilities of proliferation,migration and collagen synthesis were significantly decreased in the Doxy group,and the expression of Thy-1 and Vimentin was decreased(all P＜0.05).Conclusion:Doxy can inhibit the proliferation,migration and collagen synthesis of human corneal stromal cells induced by TGF-β1,and down-regulate the expression levels of Thy-1 and Vinmentin,thereby inhibiting corneal fibrosis.
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