Xiong Hui, Meng Fengping, Luo Miao, Chen Weiwu, Tian Juan, Chen Lunju, Ju Yankun, Mei Zhinan
School of Pharmaceutical Sciences, South-Central University for Nationalities, Wuhan, 430074, China.
Tibet Qizheng Tibetan Medicine Co.Ltd., Lasa, 850030, China.
J Ethnopharmacol. 2023 Jan 10;300:115693. doi: 10.1016/j.jep.2022.115693. Epub 2022 Sep 6.
Shi-Wei-Ru-Xiang pills (SW) as a tradition Tibetan medicine has been clinically proved effective in rheumatoid arthritis (RA) treatment. However, the underlying mechanism of SW remains unclear.
This study aimed to investigate the anti-arthritic effect of SW and its possible mechanisms of action.
A CIA rat model in vivo, and IL-1β-stimulated synoviocytes or chondrocytes and a co-culture system (IL-1β-stimulated synoviocytes/chondrocytes) in vitro were used to evaluate the effects of SW on the treatment of RA. Arthritic score, paw swelling rate, hematoxylin-eosin (HE) staining, and Safranin-O-Fast green (S-O) staining were used to evaluate the anti-arthritic activity of SW in CIA rats. TUNEL assay or flow cytometry were performed to measure chondrocytes apoptosis in vivo and invitro. The effects of SW on the expression and production of pro-inflammatory cytokines were assessed by qRT-PCR and Elisa. The inhibitory effects of SW on the phosphorylation of p38, Erk1/2, and STAT3 were analyzed by Western blot.
SW treatment significantly alleviated paw swelling, severity of arthritic and cartilage destruction in CIA rats. Moreover, SW decreased the expression of mRNAs of proinflammatory cytokines including TNF-α, IL-1β and IL-6 in the synovium, suppressed the production of these pro-inflammatory cytokines in serum and hind paws, downregulated the protein expression of p-p38, p-Erk1/2 and p-STAT3, and protected the chondrocytes apoptosis in CIA rats. Consistent with the results in vivo, SW also inhibited the activation of MAPK and STAT3 pathways, suppressed the expression of pro-inflammatory cytokines in IL-1β-stimulated synoviocytes, and attenuated chondrocytes apoptosis in IL-1β-stimulated chondrocytes. In the co-culture system, SW pre-treatment in IL-1β-stimulated synoviocytes exhibited inhibition of chondrocytes apoptosis, which was associated with attenuation of inflammation in synoviocytes.
These results suggested that the underlying mechanisms by which SW exerts its anti-arthritis effect may be related to the reduction of proinflammatory cytokine levels, inhibition of p38, Erk1/2 and STAT3 phosphorylation, and attenuating of chondrocyte apoptosis.
十味乳香丸（SW）作为一种传统藏药，临床已证明对类风湿性关节炎（RA）治疗有效。然而，SW的潜在机制仍不清楚。
本研究旨在探讨SW的抗关节炎作用及其可能的作用机制。
采用体内胶原诱导性关节炎（CIA）大鼠模型，以及体外白细胞介素-1β（IL-1β）刺激的滑膜细胞或软骨细胞和共培养系统（IL-1β刺激的滑膜细胞/软骨细胞）来评估SW对RA的治疗效果。关节炎评分、 paw肿胀率、苏木精-伊红（HE）染色和番红O-固绿（S-O）染色用于评估SW在CIA大鼠中的抗关节炎活性。采用TUNEL法或流式细胞术检测体内和体外软骨细胞凋亡情况。通过qRT-PCR和酶联免疫吸附测定（ELISA）评估SW对促炎细胞因子表达和产生的影响。通过蛋白质免疫印迹法分析SW对p38、细胞外信号调节激酶1/2（Erk1/2）和信号转导与转录激活因子3（STAT3）磷酸化的抑制作用。
SW治疗显著减轻了CIA大鼠的 paw肿胀、关节炎严重程度和软骨破坏。此外，SW降低了滑膜中包括肿瘤坏死因子-α（TNF-α）、IL-1β和IL-6在内的促炎细胞因子mRNA的表达，抑制了血清和后爪中这些促炎细胞因子的产生，下调了磷酸化p38、磷酸化Erk1/2和磷酸化STAT3的蛋白表达，并保护了CIA大鼠的软骨细胞凋亡。与体内结果一致，SW还抑制了丝裂原活化蛋白激酶（MAPK）和STAT3信号通路的激活，抑制了IL-1β刺激的滑膜细胞中促炎细胞因子的表达，并减轻了IL-1β刺激的软骨细胞中的软骨细胞凋亡。在共培养系统中，SW预处理IL-1β刺激的滑膜细胞表现出对软骨细胞凋亡的抑制作用，这与滑膜细胞炎症的减轻有关。
这些结果表明，SW发挥其抗关节炎作用的潜在机制可能与促炎细胞因子水平的降低、对p38、Erk1/2和STAT3磷酸化的抑制以及软骨细胞凋亡的减轻有关。
