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李湖帆,钟琴,马武开,朱玲,朱丹,张潇东,王敏慧,易寒知,龚文涛,刘中静,陈昌明.苗药金乌健骨胶囊对胶原诱导关节炎模型大鼠肠道、滑膜炎症及IL-17α/TRAF6/NF-κB信号通路的影响[J].湖南中医药大学学报英文版,2023,43(8):1379-1387.[Click to copy
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| 苗药金乌健骨胶囊对胶原诱导关节炎模型大鼠肠道、滑膜炎症及IL-17α/TRAF6/NF-κB信号通路的影响 |
| 李湖帆,钟琴,马武开,朱玲,朱丹,张潇东,王敏慧,易寒知,龚文涛,刘中静,陈昌明 |
| (贵州中医药大学, 贵州 贵阳 550002;贵州中医药大学, 贵州 贵阳 550002;贵州中医药大学第二附属医院风湿免疫科, 贵州 贵阳 550001;贵州医科大学附属医院临床医学研究中心, 贵州 贵阳 550001) |
| 摘要: |
| 目的 探讨苗药金乌健骨胶囊对胶原诱导关节炎（collagen-induced arthritis，CIA）模型大鼠肠道、滑膜炎症及参与NF-κB信号通路基因IL-17受体信号分子ACT1（nuclear factor kappa B activator 1，ACT1）、泛素化TRAF6（TNF receptor associated factor 6，TRAF6）、转化生长因子β激活激酶1（transforming growth factor-β-activated kinase 1，TAK1）、核因子-κB上游激酶（IκB kinase α，IKKα）、核因子κB/p65、核因子κB/p50表达的影响。方法 将70只SPF级6~8周龄雌性Wistar大鼠随机分为7组，分别为空白对照组、模型组、金乌健骨胶囊低剂量组、金乌健骨胶囊中剂量组、金乌健骨胶囊高剂量组、甲氨蝶呤组及益生菌组，每组10只。适应性喂养后除空白对照组外，其余6组构建CIA模型，两次免疫造模成功后分别进行药物金乌健骨胶囊、甲氨蝶呤、益生菌灌胃治疗，治疗4周后静脉取血，处死大鼠，分离大鼠滑膜组织及肠道组织。利用HE染色法检测大鼠滑膜及大肠组织病理情况；ELISA法检测大鼠血清TGF-β1、IL-1α、IL-17α、IL-21、IL-23的含量；Western blot法检测大鼠肠道组织ACT1、TRAF6、TAK1、IKKα、P65、P50蛋白的表达水平；RT-qPCR法检测大鼠肠道ACT1、TRAF6、TAK1、IKKα、P65、P50基因的表达情况。结果 滑膜病理结果显示，与模型组相比，金乌健骨胶囊中、高剂量组和甲氨蝶呤组炎性浸润、增生、血管新生组织有明显改善；肠道病理结果显示，与模型组相比，金乌健骨胶囊低、中、高剂量组及甲氨蝶呤组、益生菌组肠道组织细胞可见少量炎性细胞浸润，肠道组织细胞病变情况较模型组有不同程度的改善；ELISA结果显示，与模型组相比，各给药组血清含量显著降低（P<0.05或P<0.01）；Western blot结果显示，各给药组大鼠肠道组织ACT1、TRAF6、TAK1、IKKα、P65、P50的蛋白表达显著降低（P<0.05或P<0.01）；RT-qPCR结果显示，各给药组ACT1、TRAF6、TAK1、IKKα、P65、P50基因的表达显著降低（P<0.05或P<0.01）；金乌健骨胶囊高剂量组的作用最佳。结论 苗药金乌健骨胶囊能够减轻CIA大鼠肠道、滑膜炎症，同时能够影响肠道IL-17α/TRAF6/NF-κB信号途径相关基因的表达情况。 |
| 关键词: 类风湿关节炎 胶原诱导关节炎 金乌健骨胶囊 炎症 IL-17α/TRAF6/NF-κB信号通路 |
| DOI：10.3969/j.issn.1674-070X.2023.08.006 |
| Received:March 16, 2023 |
| 基金项目:国家自然科学基金项目（82260894）；国家自然科学基金项目（82060909）；贵州省科技支撑计划项目（黔科合支撑〔2020〕4Y155号）；贵州省高层次创新型人才培养计划——“百”层次人才项目（黔科合平台人才〔2016〕5650）；贵州省中医风湿免疫病临床研究中心（黔科合平台人才〔2020〕2202号）。 |
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| Effects of Jinwu Jiangu Capsule, the Miao medicine, on intestinal and synovial inflammation and IL-17α/TRAF6/NF-κB signaling pathway in rats with collagen-induced arthritis |
| LI Hufan,ZHONG Qin,MA Wukai,ZHU Ling,ZHU Dan,ZHANG Xiaodong,WANG Minhui,YI Hanzhi,GONG Wentao,LIU Zhongjing,CHEN Changming |
| (Guizhou University of Chinese Medicine, Guiyang, Guizhou 550002, China;Guizhou University of Chinese Medicine, Guiyang, Guizhou 550002, China;Department of Rheumatology and Immunology, the Second Hospital of Guizhou University of Chinese Medicine, Guiyang, Guizhou 550001, China;Research Center for Clinical Medicine, the Hospital of Guizhou Medical University, Guiyang, Guizhou 550001, China) |
| Abstract: |
| Objective To investigate the effects of Jinwu Jiangu Capsule (JWJGC), the medicine from the Chinese Miao nationality, on intestinal and synovial inflammation in rat models with collagen-induced arthritis (CIA), and on the expressions of genes involved in NF-κB signaling pathway, including IL-17 receptor signaling molecule nuclear factor kappa B activator 1 (ACT1), TNF receptor associated factor-6 (TRAF6), transforming growth factor-β-activated kinase 1 (TAK1), and the nuclear factors of IκB kinase α (IKKα), κB/p65, and κB/p50. Methods Seventy female Wistar rats aged 6-8 weeks of SPF grade were randomly divided into seven groups:blank control group, model group, low-, medium- and high-dose JWJGC groups, methotrexate group, and probiotic group, with 10 rats in each group. After adaptive feeding, except for the blank control group, CIA models were established in the other six groups. After two successful immune modeling attempts, rats in JWJGC groups, methotrexate group, and probiotic group were treated with JWJGC, methotrexate, and probiotics by gavage, respectively. After four weeks of treatment, blood of rats was collected via vein. Then the rats were sacrificed and the synovial tissues and intestinal tissues were separated from the bodies. The histopathological condition of rat synovium and intestine was checked by HE staining, the content of serum TGF-β1, IL-1α, IL-17α, IL-21, and IL-23 in rats was tested by ELISA, the expression levels of ACT1, TRAF6, TAK1, IKKα, P65, and P50 proteins in rat intestinal tissues were determined by Western blot, and the expression levels of ACT1, TRAF6, TAK1, IKKα, P65, and P50 genes in rat intestinal tissues were checked by RT-qPCR. Results The synovial pathological results showed that the medium- and high-dose JWJGC groups and methotrexate group had a significant reduction in inflammatory infiltration and proliferation, and a significant improvement in angiogenesis compared with the model group; the intestinal pathological results showed that a small amount of inflammatory cell infiltration could be seen in intestinal tissue cells of low-, medium- and high-dose JWJGC groups, methotrexate group, and probiotic group, and the pathological changes of intestinal tissue cells were improved to different degrees compared with the model group; the ELISA results showed that the content of serum TGF-β1, IL-1α, IL-17α, IL-21, and IL-23 in each medication group was significantly reduced compared with the model group (P<0.05 or P<0.01); the Western blot results showed that the expression levels of ACT1, TRAF6, TAK1, IKKα, P65, and P50 proteins in rat intestinal tissues in each medication group were significantly lower compared with the model group (P<0.05 or P<0.01); the RT-qPCR results showed that the expression levels of ACT1, TRAF6, TAK1, IKKα, P65, and P50 genes in each medication group significantly decreased compared with the model group (P<0.05 or P<0.01); the high-dose JWJGC group had the best effects (P<0.05 or P<0.01). Conclusion JWJGC, the Miao medicine, can reduce the intestinal and synovial inflammation in CIA rats, and it can affect the expressions of genes related to intestinal IL-17α/TRAF6/NF-κB signaling pathway at the same time. |
| Key words: rheumatoid arthritis collagen-induced arthritis Jinwu Jiangu Capsule inflammation IL-17α/TRAF6/NF-κB signaling pathway |
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