Effects of the fat mass and obesity-associated gene on apoptosis and the inflammatory response of chondrocytes in osteoarthritis
Objective To explore the effects of the fat mass and obesity-associated gene(FTO)on apoptosis and the inflammatory response of chondrocytes in osteoarthritis(OA).Methods Differences in FTO expression between normal human cartilage tissue samples and OA cartilage tissue samples were examined.Primary OA chondrocytes were isolated and cultured,and a rat OA model was constructed.The expression of FTO was detected in clinical,animal and cellular samples.Cells were treated with an FTO knockdown lentivirus vector(sh-FTO)and an m6A methylation inhibitor(cycloleucine).The amount of m6 A and the expression levels of inflammatory cytokines,interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α),were detected.Flow cytometry was used to detect apoptosis in OA chondrocytes,and Western blot was used to detect the expression levels of B-cell lymphoma 2(Bcl-2)and Bcl-2-associated X protein(Bax).Results Compared with the normal control group,FTO mRNA and protein expression in human OA cartilage tissue,rat OA cartilage tissue and OA chondrocytes was significantly increased(all P＜0.05).After FTO knockdown,the level of m6A increased,the levels of IL-6 and TNF-α decreased considerably,the apoptosis rate decreased,the expression of the proapoptotic protein Bax decreased considerably,and the expression of Bcl-2 increased considerably in primary OA chondrocytes.However,cycloleucine intervention clearly reduced the level of m6A,increased the levels of IL-6 and TNF-α,promoted cell apoptosis and the expression of apoptosis-related proteins,and reversed the effect induced by the FTO knockdown lentivirus in OA chondrocytes(all P＜0.05).Conclusions FTO may be involved in mechanisms related to the action of m6 A to promote OA chondrocyte apoptosis and the inflammatory response,thus accelerating the progression of OA.
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