Objective To explore the alleviatory effects of Gal-9/Tim-3 pathway on murine collagen-induced arthritis (CIA). Methods Thirty SPF female BALB/c mice (age 6-8 weeks) were used in the study. Mice model of rheumatoid arthri-tis (RA) was established by subdermal injections of collagenⅡ, on the 7th day, strengthened immunity. The day before strengthened immunity, mice were randomly divided into three groups: activation group, control group and blockage group, with 10 mice in each group, transfusion through caudal vein was performed with Gal-9, Tim-3 mAb and PBS respectively, 28 days after first immunization, the severity of CIA was evaluated with regards to arthritic index (AI), ser-a levels of IL-10, TGF-β by ELISA, expressions of biomarkers of T helper 17 cells (Th17) and regulatory T cells (Treg) by RT-PCR. Results After second immunization, the ankle joints of CIA mice began to deform with mild to se-vere arthritis. On the 28th day after first immunization, the AI of activation group was (7.12±1.53) scores, control group was (10.88±1.84) scores, and AI of blockage group was (13.82±2.12) scores, the differences were statistically significant (P<0.01). Compared with control group, the sera concentrations of IL-10 and TGF-β of activation group were signifi-cantly higher (P< 0.01) while those of blockage group were significantly lower (P< 0.01). The results of RT-PCR showed:the expression of ROR-γt in synovial tissue of activation group was lower than that of control group (P<0.01), and ROR-γt expression of blockage group was higher than that of control group (P<0.01);the expression of FoxP3 in synovial tissue of activation group was higher than control group (P< 0.01), and FoxP3 of blockage group was lower than control group (P<0.01). Conclusion Murine CIA may be markedly ameliorated by acticvation of the Gal-9/Tim-3 pathway and exacerbated by blockage of it, possibly through the tunnel of inhibiting Th17, stimulating Treg and up-regulating the expression of anti-inflammatory cytokines, which can lead to autoimmune tolerance.%目的 探讨激活或阻断Gal-9/Tim-3信号通路对小鼠胶原诱导性关节炎的影响及其机制.方法 6~8周龄雌性BALB/c小鼠30只,建立Ⅱ型胶原诱导性关节炎模型,造模后第7天加强免疫.于加强免疫前1 d随机分为激活组、阻断组、对照组,每组10只,分别经尾静脉输注半乳糖凝集素-9(Gal-9)、阻断剂Tim-3单抗、PBS.造模后28 d观察三组小鼠踝关节变形程度,记录关节炎指数,ELISA法检测血清抗炎因子白介素-10(IL-10)、转化生长因子β(TGF-β)水平,RT-PCR检测踝关节滑膜组织辅助性T细胞(Th17)、调节性T细胞(Treg)标志性蛋白ROR-γt、FoxP3表达量.结果 加强免疫后,三组小鼠出现不同程度关节变形,造模后第28天关节炎指数:激活组为(7.12±1.53)分、阻断组为(13.82±2.12)分、对照组为(10.88±1.84)分,差异有高度统计学意义(P<0.01);血清IL-10、TGF-β水平:激活组[(12.88±2.23)、(27.61±4.52)ng/L]明显高于对照组[(7.49±1.70)、(21.33±2.07)ng/L](P<0.01),阻断组[(4.26±1.79)、(12.35±2.42)ng/L]明显低于对照组(P<0.01);RT-PCR结果显示:ROR-γt mRNA在滑膜组织的表达情况激活组明显低于对照组(P<0.01),阻断组明显高于对照组(P<0.01);FoxP3 mRNA表达情况激活组明显高于对照组(P<0.01),阻断组明显低于对照组(P<0.01).结论 激活Gal-9/Tim-3通路可缓解小鼠胶原诱导性关节炎,阻断该通路则加重炎性反应,作用机制可能与该通路抑制Th17,活化Treg有关.
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