The Reversal Effect Of Wnt4 On Myofibroblasts In Oral Mucosa Scar

Abnormal activation of Wnt4 signaling has been demonstrated in the process of wound healing and the pathogenesis of fibrotic diseases.and Wnt4 is clearly identified as having a key role in the pathogenesis of renal,lung,and liver fibrosis.TGF-β1 can directly increase the expression of Wnt4 in fibro blasts.However,the specific role of Wnt4 in TGF-β 1-induced oral myofibroblast transformation and hypertrophic scar formation remains unclear.Purpose: This experiment simulated the formation of gingival scar in vitro to explore the inhibitory effect of W nt4 on the differentiation of gingival Myofibroblasts(MGF),which may represent a new therapeutic strategy for the treatment and prevention of oral mucosa scar.Methods: RNA was extracted from normal oral gingival tissue and scar tissue of oral masticatory mucosa,and the contents of Wnt4 and α-SMA were detected by q RT-PCR Human gingival fibroblasts(GF)were obtained by tissue block culture method,and identified by immunofluorescence and scanning electron microscopy.Human Gingival myofibroblasts(MGF)overexpression model,induced by TGF-β1,was used to simulate scar tissue.It was divided into three groups,namely control group,TGF-β1 group and TGF-β1+Wnt4 inhibitor group.The expression ofα-SMA was detected by RT-PCR and WB,and the effect of a small amount of Wnt4 expression on the differentiation of MGF was explored.The Wnt4 si RNA group with the highest interference efficiency was screened out.The expression ofα-SMA was detected by RT-PCR and WB methods,and the effect of Wnt4 expression on MGF differentiation was ex plored to explore the dedifferentiation ability of myofibroblasts.The above experimental data were analyzed by Graph Pad8.0 software system.Results: The contents of Wnt4 and α-SMA in scar tissue were higher than those in normal gingival tissue.After 5-8 days of culture,the cells radiated out of the tissue block,and about 90% of the cells covered the entire bottom of the containe after 15 days.Under scanning electron microscope,the cells were large and mostly star-shaped or spindle shaped.Immunofluores cence test showed positive expression of anti-vimentin and α-SMA.After Wnt4 inhibitor and Wnt4Si-RNA were applied to MGF overexpression model,RT-PCR and WB showed:Compared with the control group,the expression level of α-SMA gene in MGF over-differentiation model induced by TGF-β1 was significantly up-regulated(P<0.05),while the expression level of α-SMA gene in MGF over-differentiation model treated with Wnt4 inhibitor and Wnt4 Si-RNA was significantly inhibited(P<0.05).Conclusion: The expression lev els of Wnt4 and α-SMA in scar tissue of oral mucosa were higher than those in normal mucosa,and sufficient cells were obtained by tissue block culture method to be verified as GF.TGF-β1 could induce MGF differentiation model can be established to simulat e scar formation,while Wnt4 could inhibit the differentiation of MGF induced by TGF-β1 and reduce the contraction activity of MGF.