基于Tim-3/Gal-9信号通路探讨艾灸对类风湿关节炎大鼠的治疗作用OA
Therapeutic effects of moxibustion on rheumatoid arthritis rats based on Tim-3/Gal-9 signaling pathway
目的:基于 T 细胞免疫球蛋白黏蛋白域蛋白3/半乳凝素-9(Tim-3/Gal-9)信号通路探讨艾灸对类风湿关节炎(RA)大鼠的治疗作用.方法:24 只 SD 大鼠随机分为对照组、模型组、艾灸组、干扰+艾灸组,每组6 只.于大鼠两后足足跖皮下注射100 μL 0.5 mg/mL 弗氏完全佐剂建立 RA 模型,7 d 后艾灸组每日于肾俞(BL23)、足三里(ST36)穴艾灸5 次,干扰+艾灸组于两后足足垫注射 sh-Tim-3 慢病毒表达载体,1 h 后进行艾灸,每周治疗 6 d,持续3 周.干预结束后,测量大鼠足垫厚度,ELISA 法检测血清 TNF-α、IL-6、IL-12、IL-13、TGF-β 水平,HE 染色观察踝关节滑膜组织学变化并进行病理评分,免疫荧光检测滑膜巨噬细胞表型,实时荧光定量 PCR 检测脾脏、滑膜组织中Tim-3、Gal-9 mRNA 表达,Western blot 检测滑膜组织中 Tim-3、Gal-9 蛋白表达.结果:与对照组相比,模型组左、右足第7、14、21、28 天足垫厚度,血清 TNF-α、IL-6、IL-12 水平,滑膜组织中 iNOS+/CD206+比值升高;血清 IL-13、TGF-β 水平,脾脏组织中 Tim-3、Gal-9 mRNA 及滑膜组织中 Tim-3、Gal-9 mRNA 和蛋白表达水平下降(P＜0.05).与模型组相比,艾灸组上述指标变化趋势相反(P＜0.05).慢病毒干扰可削弱艾灸的作用(P＜0.05).结论:艾灸可能通过调控 Tim-3/Gal-9 信号通路调节巨噬细胞极化,增强抗炎能力,缓解 RA 大鼠临床症状.
Aim:To investigate the therapeutic effects of moxibustion on rheumatoid arthritis(RA)rats based on the T cell immunoglobulin and mucin domain-containing protein 3/galectin-9(Tim-3/Gal-9)signaling pathway.Methods:Twen-ty-four SD rats were randomly divided into control group,model group,moxibustion group,and sh-Tim-3+moxibustion group,with 6 rats in each group.RA was induced by subcutaneously injecting 100 μL of 0.5 mg/mL Freund's complete ad-juvant into the foot pads of both hind paws.Seven days later,rats in the moxibustion group received moxibustion at Shenshu(BL23)and Zusanli(ST36)acupoints 5 times daily.The sh-Tim-3+moxibustion group was injected with sh-Tim-3 lentivir-us expression vector into the bilateral hind foot pads,and moxibustion was performed 1 hour later.All treatments performed 6 days per week for 3 weeks.After the intervention,the foot pad thickness was measured.Serum levels of TNF-α,IL-6,IL-12,IL-13,and TGF-β were detected by ELISA.Histological changes of the ankle synovial tissue were observed by HE stai-ning,and pathological scoring was performed.Macrophage phenotypes in the synovial tissue were assessed by immunofluo-rescence.Tim-3 and Gal-9 mRNA expression in the spleen and synovial tissues were detected by real-time quantitative PCR.Tim-3 and Gal-9 protein expression in the synovial tissue were detected by Western blot.Results:Compared with the control group,the model group exhibited increased footpads thickness on days 7,14,21 and 28,elevated serum levels of TNF-α,IL-6,and IL-12,and a higher synovial iNOS+/CD206+ratio(P＜0.05).Conversely,serum levels of IL-13 and TGF-β,Tim-3 and Gal-9 mRNA expression in the spleen and synovial tissues,as well as Tim-3 and Gal-9 protein expression in the synovial tissue decreased(P＜0.05).Compared with the model group,the changes in the above indexes of the moxi-bustion group were opposite(P＜0.05).The lentivirus weakened the effects of moxibustion(P＜0.05).Conclusion:Moxi-bustion may regulate macrophage polarization through the Tim-3/Gal-9 signaling pathway,enhance the anti-inflammatory ca-pacity,and alleviate the clinical symptoms of RA rats.
徐溢梅;段玉松;聂敬伟;黄俊;崔露;黄志灿;周海燕
成都中医药大学针灸推拿学院 成都 610041成都中医药大学针灸推拿学院 成都 610041成都中医药大学针灸推拿学院 成都 610041成都中医药大学针灸推拿学院 成都 610041成都中医药大学针灸推拿学院 成都 610041成都中医药大学针灸推拿学院 成都 610041成都中医药大学针灸推拿学院 成都 610041
医药卫生
T细胞免疫球蛋白黏蛋白域蛋白3半乳凝素-9艾灸类风湿关节炎大鼠
T cell immunoglobulin and mucin domain-containing protein 3galectin-9moxibustionrheumatoid arthri-tisrat
《郑州大学学报（医学版）》 2026 (3)
29-33,5
国家自然科学基金项目(82374587,81973959)四川省自然科学基金项目(2024NSFSC2113)
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