Objective:To investigate the effect of Wu-tou decoction on the expressions of matrix metalloproteinase 13(MMP-13)and tumor necrosis factor-α(TNF-α)in osteoarthritis(OA)chondrocytes cell,and its regulatory mechanism on silent information regulator 1(SIRT1)/forkhead box transcription factor O1(FOXO1)signaling pathway.Methods:Lipopolysaccharide was used to simulate inflammatory microenvironment to construct osteoarthritis chondrocyte model.EX527,SIRT1 signal specific inhibitor,was used to perform functional rescue experiment.Cell experiments were divided into normal control group,lipopolysaccharide group,low dose group of Wu-tou decoction,high dose group of Wu-tou decoction and inhibitor group.Calcein AM/propidium iodide(PI)staining was used to detect the growth activity of cells in each group.Flow cytometry was used to detect the apoptosis rate.Immunofluorescence was used to detect the expression of TNF-α and MMP-13.Immunohistochemistry was used to detect the expression of Aggrecan and Collagen Ⅱ.Western Blot was used to detect the expression of SIRT1,acetylated FOXO1(Ac-FOXO1),B cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)in cells.Results:In osteoarthritis cells,the growth activity of cells decreased significantly,the apoptosis rate increased significantly,the expressions of TNF-α and MMP-13,Ac-FOXO1 and Bax were increased signifi-cantly,and the expressions of Aggrecan and Collagen Ⅱ,SIRT1 and Bcl-2 were decreased significantly.Low and high dose of Wu-tou decoction could significantly block the deterioration of these parameters and exert a protective effect of chondro-cytes,while EX527 could partially reverse the protective effect of Wu-tou decoction.The differences were statistically sig-nificant(P＜0.05).Conclusion:Wu-tou decoction could significantly down-regulate the expression of TNF-α and MMP-13 and inhibit the apoptosis of chondrocytes in osteoarthritis,which may be related to the activation of SIRT1 signal and inhi-bition of FOXO1 acetylation.
维普
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