Extract from Eucommia ulmoides Oliv. ameliorates arthritis via regulation of inflammation, synoviocyte proliferation and osteoclastogenesis in vitro and in vivo
Extract from Eucommia ulmoides Oliv. ameliorates arthritis via regulation of inflammation, synoviocyte proliferation and osteoclastogenesis in vitro and in vivo
复制标题
杜仲提取物。
DOI：
10.1016/j.jep.2016.10.038
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发表时间：
2016-12-24
影响因子：
5.4
通讯作者：
Yang, Yong-Qing
中科院分区：
文献类型：
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作者：
Wang, Jian-Ying;Yuan, Ying;Yang, Yong-Qing
民族药理学相关性：杜仲，一种珍贵的中草药，已经显示出多种健康益处。尽管中药在临床上广泛用于治疗类风湿性关节炎（RA），但很少有研究描述其在RA中的抗病理作用或机制。本研究探讨了杜仲提取物的作用机理。材料与方法：观察四种不同杜仲提取物对胶原诱导性关节炎（CIA）大鼠模型的影响。采用MTT法检测提取物对类风湿关节炎成纤维样滑膜细胞（RA FLS）增殖的影响，流式细胞仪检测提取物对RA FLS细胞凋亡的影响。其中，70%乙醇提取物（EU 70）表现出最好的抑制作用，并进一步研究其对CIA大鼠的疗效。检测足肿胀，并对关节炎指数（AI）进行评分。通过关节组织的苏木精和伊红（H&E）染色评估病理学改善。EU 70的机制作用研究如下：通过流式细胞术检测的Th 17阳性细胞中的抗炎作用;通过ELISA检测的炎性细胞因子的血清水平;通过免疫组织化学（IHC）检测的TNF α和IL-1 β表达;以及通过RANKL和OPG mRNA的QPCR检测的抗破骨细胞生成。与溶剂组相比，EU 70能显著改善CIA大鼠足肿胀，降低AI，减少炎症细胞浸润和滑膜细胞增殖。EU 70降低了脾脏中Th 17阳性细胞的数量和细胞因子（包括IL-17、IL-1 β和TNF α）的血清水平，并上调了IL-10的血清水平;这些结果表明了EU 70的抗炎作用。此外，EU 70还能有效抑制关节组织中TNF α和IL-1 β的表达，并导致RANK mRNA的下调和OPG mRNA的上调。这些结果揭示了EU 70对骨破坏的可能预防作用。结论：首次，这些机制和病理学的改善支持杜仲的临床使用。治疗RA。结果表明，杜仲70%乙醇提取物具有较好的抑菌效果。可以通过以下方式缓解RA症状：（1）抑制滑膜细胞增殖，（2）减少Th 17阳性细胞数量并下调血清IL-17表达，（3）增强IL-10的抗炎作用，（4）抑制关键促炎细胞因子TNF α和IL-1 β的血清和组织水平，以及（5）减少软骨和骨的降解。
Ethnopharmacological relevance: Eucommia ulmoides Oliv., a valuable Chinese herb, has shown a variety of health benefits. Despite the widespread clinical use of this herb to treat rheumatoid arthritis (RA) in Traditional Chinese Medicine (TCM), very few studies have described its anti-pathological effects or mechanism in RA. The present study investigated the mechanism of Eucommia ulmoides Oliv. in an experimental collagen-induced arthritis (CIA) rat model.Materials and methods: The effects of four different Eucommia ulmoides Oliv. extracts on the proliferation of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) were screened in an MTT assay, and apoptotic effects were detected by flow cytometry. Among the extracts, the 70% ethanol extract (EU70) presented the best inhibition and was further investigated for its curative effect in CIA rats. Foot swelling was detected, and the arthritis index (AI) was scored. Pathological improvement was assessed by haematoxylin and eosin (H&E) staining of joint tissues. The mechanistic effects of EU70 were investigated as follows: anti-inflammatory effects in Th17-positive cells by flow cytometry; serum levels of inflammatory cytokines by ELISA; TNF alpha and IL-1 beta expression by immunohistochemistry (IHC); and anti-osteoclastogenesis by QPCR detection of RANKL and OPG mRNA.Results: Compared with vehicle treatment in CIA model rats, EU70 significantly ameliorated foot swelling, decreased AI in vivo and reduced inflammatory cell infiltration and synoviocyte proliferation. EU70 decreased the number of Th17-positive cells in the spleen and the serum levels of cytokines, including IL-17, IL-1 beta and TNF alpha, and upregulated the serum levels of IL-10; these results indicated the anti-inflammatory effect of EU70. Moreover, EU70 effectively suppressed TNF alpha and IL-1 beta expression in the joint tissues and resulted in the downregulation of RANK', mRNA and the upregulation of OPG mRNA. These results revealed the possible preventive role of EU70 against bone destruction.Conclusion: For the first time, these mechanisms and pathological improvements support the clinical use of Eucommia ulmoides Oliv. in treating RA. The findings indicated that the 70% ethanol extract of Eucommia ulmoides Oliv. could relieve RA symptoms by (1) suppressing the proliferation of synoviocytes, (2) reducing the number of Th17-positive cells and downregulating serum IL-17 expression, (3) increasing the anti-inflammatory effects of IL-10, (4) inhibiting the serum and tissue levels of key pro-inflammatory cytokines, TNF alpha and IL-1 beta, and (5) reducing the degradation of cartilage and bone.