目的 研究通痹颗粒对胶原诱导性关节炎(collagen-induced arthritis,CIA)大鼠铁调素(hepcidin,Hepc)、Janus激酶(janus kinase,JAK)2/信号转导子和转录激活子(signal transduction and activator of transcription,STAT)3 信号通路的影响.方法 选取36只雌性SD大鼠随机分成空白组、模型组、阳性对照组和通痹颗粒低、中、高剂量组,每组6只.空白组不予处理,其余组用牛Ⅱ型胶原建立CIA模型.造模完成后,空白组、模型组予生理盐水灌胃,其余各组分别以巴瑞替尼片和低、中、高剂量通痹颗粒灌胃.每天1次,连续4周.HE染色行滑膜组织病理学观察;酶联免疫吸附法测定血清Hepc、白细胞介素6(interleukin 6,IL-6)水平;逆转录-聚合酶链反应法测定滑膜中JAK2、STAT3、细胞信号因子传导抑制体(suppressor of cytokine signaling,SOCS)1、SOCS3的mR-NA相对表达量;Western blot法检测滑膜中JAK2、p-JAK2、STAT3、p-STAT3、SOCS1、SOCS3的蛋白表达量.结果 模型组见滑膜上皮结构缺损,滑膜重度增生,排列紊乱,并有大量炎症细胞浸润和多个血管翳形成;各给药组滑膜炎症均有所减轻,阳性对照组优于通痹颗粒高剂量组,通痹颗粒中、高剂量组优于低剂量组.与模型组相比,各给药组关节炎指数评分、血清Hepc和IL-6水平均显著降低(P＜0.01);与阳性对照组相比,通痹颗粒中、低剂量组关节炎指数评分、血清Hepc和IL-6水平均升高(P＜0.05).与模型组比较,阳性对照组和通痹颗粒低、中、高剂量组JAK2、STAT3 mRNA和蛋白以及p-JAK2、p-STAT3的蛋白表达量均降低(P＜0.05),而通路抑制因子SOCS1、SOCS3 mRNA和蛋白的表达均升高(P＜0.05);与阳性对照组比较,通痹颗粒各剂量组JAK2、STAT3 mRNA和蛋白以及p-JAK2、p-STAT3的蛋白表达量均升高(P＜0.05),而SOCS1、SOCS3 mRNA和蛋白的表达均降低(P＜0.05).结论 通痹颗粒能够改善CIA大鼠滑膜炎症,其机制可能与抑制JAK2/STAT3信号通路而减少Hepc的表达有关.
Effects of Tongbi Granule on collagen-induced arthritis rats based on Hepcidin and JAK2/STAT3 signaling pathway
Objective To study the effects of Tongbi Granule on hepcidin(Hepc)and janus kinase(JAK)2/signal transduction and activator of transcription(STAT)3 signaling pathway in collagen-induced arthritis(CIA)rats.Methods A total of 36 female SD rats were randomized into blank,model,positive control,low-,medium-,and high-dose of Tongbi Granule groups,with six rats in each group.CIA model was constructed with bovine type Ⅱ collagen in all groups except the blank group.After modeling,the blank and model groups were given normal saline by gavage,the other groups were given baricitinib tablets,and low-,medium-,and high-dose of Tongbi Granule by gavage respectively,once a day,for continual four weeks.HE staining was performed to observe pathological synovial tissue.Serum Hepc and interleukin 6(IL-6)levels were measured by ELISA.The mRNA relative expressions of JAK2,STAT3,suppressor of cytokine signaling(SOCS)1,and SOCS3 in synovium were determined by RT-PCR.Western blot was used to test the protein expressions of JAK2,p-JAK2,STAT3,p-STAT3,SOCS1 and SOCS3 in synovium.Results In the model group,there were defects of synovial epithelium,severe synovial hyperplasia,disordered arrangement,and a large number of inflammatory cell infiltration and pannus formation.However,all administration groups showed reduction of synovial inflammation,the positive control group was better than the high-dose Tongbi Granule group,and the medium-and high-dose groups was better than the low-dose group.Compared with model group,arthritis index scores,and levels of serum Hepc and IL-6 were significantly lower in all drug administration groups(P＜0.01).Compared with the positive control group,the arthritis index scores,serum Hepc and IL-6 levels in low-and medium-dose Tongbi Granule groups were higher(P＜0.05).Compared with model group,the mRNA and protein expressions of JAK2 and STAT3,as well as the protein expressions of p-JAK2 and p-STAT3 were lower(P＜0.05),while the mRNA and protein expressions of pathway inhibitors SOCS1 and SOCS3 were higher in positive control group and low-,medium-,and high-dose Tongbi Granule groups(P＜0.05).Compared with positive control group,the mRNA and protein expressions of JAK2 and STAT3,and the protein expressions of p-JAK2 and p-STAT3 were higher in Tongbi Granule groups of various doses(P＜0.05),while the mRNA and protein expressions of SOCS1 and SOCS3 were lower(P＜0.05).Conclusion Tongbi Granule can relieve synovial inflammation in CIA rats,and its mechanism may be related to inhibiting JAK2/STAT3 signaling pathway and reducing the expression of Hepc.
rheumatoid arthritiscollagen-induced arthritisChinese medicinesTongbi Granulehepcidinjanus kinase 2/signal transduction and activator of transcription 3 signaling pathway
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