The Effect Of TGF-β1 Inhibitor Oxymatrine On Biological Characteristics Of Pterygium Fibroblasts And Its Mechanism

Objective:To investigate the effect of oxymatrine(OM),an inhibitor of transforming growth factor betal(TGF-β1),on the proliferation,migration and invasion of human pterygium fibroblasts(HPFs),and to explore its mechanism,so as to provide a new direction for prevention,treatment and prevention of postoperative pterygium recurrence.Methods:1.Normal fibroblasts(NFs)and HPFs were obtained by primary culture.The morphology of the cells was observed and the cells were identified by ICC method.2.CCK-8 assay,wound healing assay and Transwell cell invasive assay were used to detect the proliferation,migration and invasion ability of NFs and HPFs.3.RT-qPCR and WB were used to detect the expression of TGF-β1 and EMT related indexes E-cadherin and vimentin mRNA and protein in NFs and HPFs.4.CCK-8 assay was used to detect the effects of different concentrations and the same concentration with different actuation durationof OM on the proliferation of HPFs.5.HPFs were treated with OM of suitable concentration and divided into untreated group(HPFs group)and treated group(OM group).Cell migration and invasion ability,mRNA and protein changes of TGF-β1 and EMT related indexes were detected between the two groups.Result:1.NFs and HPFs were cultured,purified and identified successfully.Vimentin was positive and cytokeratin was negative in both of them.Combined with their cell morphology,they could be identified as fibroblasts.2.CCK-8,wound healing assay and Transwell cell invasive assayshowed that the proliferation,migration and invasion ability of HPFs were stronger than that of NFs,and the difference was statistically significant(P<0.05).3.RT-qPCR and WB results showed that the mRNA and protein expressions of TGF-β1 and vimentin in HPFs were higher than those in NFs,while E-cadherin was lower than that in NFs(P<0.05).4.OM could inhibit the proliferation of HPFs,and the optimal concentration and time were 4mg/ml and 48h.5.The mRNA and protein expressions of TGF-β1 and vimentin in OM group were lower than those in HPFs group,while E-cadherin was higher than those in HPFs group,and the differences were statistically significant(P<0.05).In addition,the migration and invasion ability of OM group was weaker than that of HPFs group,and the difference was statistically significant(P<0.05).Conclusion:1.There are significant differences in morphology,proliferation,migration and invasion between NFs and HPFs.It is speculated that the biological characteristics of fibroblasts in pterygium have changed,which may be related to TGF-β1 and EMT.2.OM inhibited the proliferation,migration and invasion of HPFs in a time and concentration dependent manner.Its effect is closely related to the partial down-regulation of TGF-β1 and the further reversal of EMT related indicators.It may become a neoadjuvant drug for the prevention,treatment and prevention of postoperative recurrence of pterygium,but its specific mechanism needs to be further studied.