Tibetan medicine Kuan-Jin-Teng exerts anti-arthritic effects on collagen-induced arthritis rats via inhibition the production of pro-inflammatory cytokines and down-regulation of MAPK signaling pathway
Tibetan medicine Kuan-Jin-Teng exerts anti-arthritic effects on collagen-induced arthritis rats via inhibition the production of pro-inflammatory cytokines and down-regulation of MAPK signaling pathway
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藏药宽金藤通过抑制促炎细胞因子的产生、下调MAPK信号通路对胶原性关节炎大鼠发挥抗关节炎作用
DOI：
10.1016/j.phymed.2018.12.023
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发表时间：
2019-04-01
期刊：
影响因子：
7.9
通讯作者：
Mei Zhinan
中科院分区：
文献类型：
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作者：
Xiong Hui;Ding Xin;Mei Zhinan
背景：中华青牛胆（Tinospora sinensis (Lour.) Merr）的茎在中文中俗称“宽金藤”，已作为藏药用于治疗类风湿性关节炎。目的：在胶原诱导性关节炎（CIA）模型中研究中华青牛胆（Tinospora sinensis (KJT)）茎乙醇提取物的 EtOAc 部分对促炎细胞因子和 MAPK 通路的影响。研究设计：KJT 的抗关节炎活性方法：采用LC-MS和HPLC对KJT的化学成分进行分析。采用弗氏佐剂乳化牛CII注射Wistar大鼠建立CIA模型。每天通过口服强饲法给 CIA 大鼠施用几个剂量的 KJT（50.0、100.0 和 200.0 mg/kg），持续 4 周。通过临床关节炎评分、爪肿胀检查和痛觉过敏测量以及 CIA 大鼠的放射学和组织学分析来研究 KJT 的抗关节炎活性。分别通过蛋白质印迹、免疫组化染色、ELISA和实时荧光定量PCR检测KJT对CIA大鼠踝关节、血清和脾脏中MAPK通路激活和促炎细胞因子（TNF-α、IL-1β和IL-17）产生的影响。最后，在脂多糖（LPS）刺激的RAW264.7小鼠巨噬细胞中检测了康健汤对一氧化氮（NO）和促炎细胞因子的产生以及对p38和Erk磷酸化的调节的影响。结果：康健汤显着减轻了CIA大鼠的足爪肿胀、痛觉过敏和关节炎的严重程度，并减少了滑膜组织增殖和炎症细胞浸润。此外，KJT 抑制 CIA 大鼠踝关节、血清和脾脏中 TNF-α、IL-1β 和 IL-17 的产生，并逆转 p38 和 Erk 磷酸化的上调。 KJT 还被证明可以抑制 LPS 刺激的 RAW264.7 细胞中 NO 和促炎细胞因子（TNF-α、IL-1β 和 IL-6）的产生以及 p38 和 Erk 的磷酸化。结论：这些结果表明 KJT 发挥抗关节炎作用的机制可能归因于抑制促炎细胞因子的产生和下调 MAPK 信号通路。
Background: The stems of Tinospora sinensis (Lour.) Merr commonly named "Kuan-Jin-Teng" in Chinese, have been used to treat rheumatoid arthritis as a Tibetan medicine.Purpose: The effects of the EtOAc fraction of ethanolic extract from the stems of T. sinensis (KJT) on the proinflammatory cytokines and MAPK pathway were studied in collagen-induced arthritis (CIA) model.Study design: Anti-arthritic activity of KJT was investigated in CIA model.Methods: The chemical constituents of KJT were analyzed by LC-MS and HPLC. The CIA model was established with injecting the bovine CII emulsified in Freund's adjuvant in Wistar rats. Several doses of KJT (50.0, 100.0 and 200.0 mg/kg) were administrated via oral gavage to CIA rats daily for 4 weeks. The anti-arthritic activity of KJT was investigated by clinical arthritis scoring, paw swelling inspection and hyperalgesia measurement, as well as radiological and histological analysis in CIA rats. The impacts of KJT on the activation of MAPK pathway, production of pro-inflammatory cytokines (TNF-alpha, IL-1 beta and IL-17) in ankle joints, serum, and spleen in CIA rats were examined by western blot, immunohistochemical staining, ELISA, and quantitative real-time PCR respectively. Lastly, the effects of KJT on production of the nitric oxide (NO) and pro-inflammatory cytokines as well as the regulation of the phosphorylation of p38 and Erk were detected in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage cells.Results: KJT significantly alleviated the paw swelling, hyperalgesia and arthritic severity, and reduced the synovial tissue proliferation and inflammatory cell infiltration in the CIA rats. Moreover, KJT suppressed the production of TNF-alpha, IL-1 beta, and IL-17 in ankle joints, serum, and spleen and reversed the up-regulation of the phosphorylation of p38 and Erk in CIA rats. KJT was also demonstrated to inhibit the production of NO and proinflammatory cytokines (TNF-alpha, IL-1 beta and IL-6), and phosphorylation of p38 and Erk in LPS-stimulated RAW264.7 cells.Conclusion: These results suggest the mechanisms of KJT performing its anti-arthritis effect may be attributed to inhibiting the production of pro-inflammatory cytokines and down-regulating the MAPK signaling pathway.