Chen Yifei, Liu Kaifei, Qin Yingyuan, Chen Suyi, Guan Guokai, Huang Yao, Chen Yu, Mo Zhixian
School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China.
School of Pharmacy, Guilin Medical University, Guilin, China.
Front Pharmacol. 2022 May 9;13:869810. doi: 10.3389/fphar.2022.869810. eCollection 2022.
To investigate the therapeutic effect of petroleum ether extract of Miller (PEEP) on rheumatoid arthritis (RA). : The Cell Counting Kit-8 (CCK-8) was used to detect cell activity and select the optimal concentration of the extract; the effective site was screened by nitric oxide (NO) colorimetric method and Q-PCR method; the expression of p38, p-p38, p-MK2, and Tristetraprolin (TTP) in RAW 264.7 cells were detected by Western blot. : The rat model was established by complete Freund's adjuvant (CFA). The different doses of PEEP on CFA rats were observed with life status, paw swelling, spleen index, X-ray, Hematoxylin eosin (HE) staining; the secretion of Tumor necrosis factor α (TNF-α), interleukin-6 (IL-6) and Prostaglandin E2 (PGE) were detected by Enzyme linked immunosorbent assay (ELISA); the expressions of p38, p-p38, p-MK2, and TTP in the ankle joints of CFA rats were detected by Western blot. : PEEP, Ethyl Acetate Extract of acul Miller (EEEP), N-butanol Extract of Miller (BEEP) have no toxic effects on RAW264.7 macrophages. PEEP, EEEP, and BEEP reduce the secretion of NO in RAW264.7 cells induced by lipopolysaccharide (LPS), only PEEP significantly inhibited the mRNA expression levels of inflammatory factors TNF-α and IL-6; PEEP-dependently reduce the secretion of TNF-α and IL-6, decrease the expression of p-p38 and p-MK2, and the level of TTP phosphorylation in LPS-induced RAW264.7 cells. : PEEP improve the living conditions of CFA rats, reduce foot swelling, spleen index, bone surface erosion and joint space narrowing; reduce the formation of synovial cells, inflammatory cells and pannus in the foot and ankle joints. PEEP reduce the secretion of TNF-α, IL-6, PGE in rat serum, downregulate the expression of p-p38 and p-MK2 in the ankle joint, and reduce the phosphorylation of TTP. PEEP improve the living conditions of CFA rats, reduce the degree of foot swelling, protect immune organs, reduce inflammatory cell infiltration, cartilage damage, pannus formation, reduce inflammation and RA damage. The mechanism through regulating the signal pathway of p38 mitogen-activated protein kinase (p38/MAPK), which reduces the release of TNF-α, IL-6, and PGE in the serum.
探讨米勒石油醚提取物（PEEP）对类风湿关节炎（RA）的治疗作用。：采用细胞计数试剂盒-8（CCK-8）检测细胞活性并筛选提取物的最佳浓度；通过一氧化氮（NO）比色法和Q-PCR法筛选有效部位；采用蛋白质免疫印迹法检测RAW264.7细胞中p38、p-p38、p-MK2和三联四肽重复蛋白（TTP）的表达。：采用完全弗氏佐剂（CFA）建立大鼠模型。观察不同剂量的PEEP对CFA大鼠的生命状态、 paw肿胀、脾脏指数、X射线、苏木精-伊红（HE）染色的影响；采用酶联免疫吸附测定（ELISA）检测肿瘤坏死因子α（TNF-α）、白细胞介素-6（IL-6）和前列腺素E2（PGE）的分泌；采用蛋白质免疫印迹法检测CFA大鼠踝关节中p38、p-p38、p-MK2和TTP的表达。：PEEP、米勒乙酸乙酯提取物（EEEP）、米勒正丁醇提取物（BEEP）对RAW264.7巨噬细胞无毒性作用。PEEP、EEEP和BEEP可降低脂多糖（LPS）诱导的RAW264.7细胞中NO的分泌，仅PEEP显著抑制炎症因子TNF-α和IL-6的mRNA表达水平；PEEP依赖性地降低TNF-α和IL-6的分泌，降低p-p38和p-MK2的表达以及LPS诱导的RAW264.7细胞中TTP的磷酸化水平。：PEEP改善CFA大鼠的生活状况，减轻足肿胀、脾脏指数、骨表面侵蚀和关节间隙变窄；减少足和踝关节滑膜细胞、炎症细胞和血管翳的形成。PEEP降低大鼠血清中TNF-α、IL-6、PGE的分泌，下调踝关节中p-p38和p-MK2的表达，降低TTP的磷酸化。PEEP改善CFA大鼠的生活状况，减轻足肿胀程度，保护免疫器官，减少炎症细胞浸润、软骨损伤、血管翳形成，减轻炎症和RA损伤。其机制是通过调节p38丝裂原活化蛋白激酶（p38/MAPK）信号通路，从而减少血清中TNF-α、IL-6和PGE的释放。
