Effects of CD300c-Ig on joint injury in mice with collagen-induced arthritis
Objective To investigate the effects and mechanism of CD300c-Ig on joint injury in collagen-induced arthri-tis(CIA)mice.Methods To construct the CIA model,DBA/1J mice were immunized on day 1 and day 21,and then randomly divided into the treatment group and control group,with 10 mice in either group.Mice in the treatment group were intraperitoneally injected with 100 μL of 20 μg CD300c-Ig once every 3 days for 6 times,with a total dose of 120 μg;mice in the control group were intraperitoneally injected with control Ig of the same volume once every 3 days for 6 times.The degree of swelling in the joints of the extremities was observed and the arthritis index score was re-corded.Forty days after the initial immunization,the spleens of the mice were taken to make single-cell suspension,and the expressions of T regulatory cells(Treg),T helper cells(Th17),tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ)were detected with flow cytometry.The lower limbs of the mice were taken,decalcified,and sliced for he-matoxylin-eosin staining and safranin O staining to visualize the pathological features of the joints and to score them.The mRNA expressions of forkhead box protein P3(Foxp3),interleukin-17(IL-17)and interleukin-23(IL-23)were detected with fluorescence quantitative PCR.Results The degree of joint swelling and AI in the treatment group were significantly reduced compared with those in the control group.Hematoxylin-eosin staining and saffron O staining showed that the proliferation of synoviocytes in the synovial joints of the treatment group was reduced,and the forma-tion of vascular cataracts and the erosion of bone were significantly improved(P<0.05).Flow cytometry showed that the proportion of Th17 cells in the spleens of the treatment group was significantly lower(P<0.05)while the proportion of Treg cells was significantly higher(P<0.05).The expressions of TNF-α and IFN-γ in the spleen of the treatment group were significantly lower(P<0.05).Fluorescence quantitative PCR showed that the mRNA expressions of IL-17 and IL-23 in the treatment group were significantly lower.Conclusion CD300c-Ig can reduce joint injury in CIA mice,and the mechanism is related to its regulation of Th17/Treg cell balance and inhibition of inflammatory factor release.
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