奇壬醇通过Wnt5a/FZD5通路抗类风湿性关节炎的机制研究OA

Objective:To investigate the inhibitory effects of kirenol on collagen-induced arthritis(CIA)in rats and transforming growth factor-beta 1(TGF-β1)-induced proliferation of human rheumatoid arthritis synovial fibroblasts(MH7A)and explore the associated mechanisms.Methods:(1)The CIA model rats were randomly grouped as follows:model,kirenol(1.25,2.5,and 5 mg/kg),and prednisone(21 mg/kg),and a normal control group was set up.The rats were administrated with corresponding drugs by gavage at 10 mL/kg body mass,once a day for 14 days.The arthritis in-dex was adopted to evaluate the degree of joint inflammation.A toe volume measuring instrument was used to measure the toe volume and calculate the toe swelling degree.Hematoxylin-eosin staining and safranin O-fast green staining were conducted to observe the pathological changes of the synovial tissue and the bone tissue of the ankle joint.The serum lev-els of rheumatoid factor(RF),tumor necrosis factor(TNF)-α,interleukin(IL)-10,and IL-17 were determined by en-zyme-linked immunosorbent assay(ELISA).(2)MH7A cells were treated with kirenol at different concentrations.After 24 h,the cell proliferation was detected by the CCK-8 method.The cells treated with 10 ng/mL TGF-TGF-β1 for 24 h were allocated into control,kirenol(2.5,5,and 10 μmol/L),and positive control groups,and MH7A cells were selected as the normal control group.After kirenol intervention,the cell proliferation rate was measured by the CCK-8 method,and the migration of cells was examined by the scratch test.The content of TNF-α in the cell supernatant was measured by ELISA.The nucleus-specific expression of nuclear factor(NF)-κB p65 in the nucleus was observed by immunofluores-cence.The expression of wingless-type mouse mammary tumor virus integration site family,member 5a(Wnt5a)/frizzled class receptor 5(FZD5)pathway-related proteins was determined by Western blot.Results:In the animal experiment,compared with the control group,the model group showed increases in arthritis index,toe swelling,and serum levels of RF,TNF-α,and IL-17,a decrease in the serum level of IL-10,and obvious pathological changes.In the cell experiment,compared with the control group,the model group showed enhanced cell proliferation and migration,up-regulated protein levels of Wnt5a,FZD5,and PKC and phosphorylation level of IκB,and increased nuclear expression of NF-κB p65 and release of TNF-α(P<0.01).Compared with the model group,kirenol reduced the arthritis index,toe swelling,and ser-um levels of RF,TNF-α,and IL-17,and increased the serum level of IL-10(P<0.05 or 0.01).In addition,kirenol alle-viated the pathological changes such as abnormal proliferation and inflammatory cell infiltration of the synovial tissue,in-vasion of the synovial tissue to the cartilage tissue,and loss of the cartilage matrix in CIA rats.Kirenol inhibited abnormal cell proliferation and migration,down-regulated the protein levels of Wnt5a,FZD5,and PKC and the phosphorylation lev-el of IκB,and reduced the nuclear expression of NF-κB p65 and the release of TNF-α(P<0.05 or 0.01).Conclusion:Kirenol has a significant therapeutic effect on CIA in rats.It can ameliorate the abnormal proliferation and inflammatory cell infiltration of the synovial tissue and significantly inhibit the proliferation and migration of MH7A cells by down-regu-lating the activation of the Wnt5a/FZD5 pathway and inhibiting the release of pro-inflammatory cytokines.