基于内质网应激通路探讨金铁锁醇提物治疗类风湿性关节炎的机制

Exploring the mechanism of golden Psammosilene tunicoides extract in treating rheumatoid arthritis based on the endoplasmic reticulum stress pathway

关键词:  类风湿性关节炎  金铁锁醇提物  CIA模型  内质网应激  炎症

Key words:  rheumatoid arthritis  CEPT  CIA model  endoplasmic reticulum stress  inflammation

摘要:

云南民间治疗风湿痹症常用药金铁锁在现代用于研究治疗RA,基于内质网应激通路进行体内实验,探索金铁锁醇提物(CEPT)的作用机制。方法:采用胶原诱导的类风湿性关节炎(CIA)小鼠模型,按照各给药组剂量连续给药21天。进行足肿胀和关节炎评分、组织病理学染色；酶联免疫吸附法(ELISA)检测小鼠血清中肿瘤坏死因子-α(TNF-α)的含量；蛋白印迹 (Western blot) 法测定小鼠踝关节中转录激活因子6(ATF-6)、葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)蛋白的表达；实时荧光定量核酸扩增(real-time quantitative PCR,q-PCR检测细胞中ATF-6、GRP78、CHOP的mRNA表达。结果:各给药组显著降低了CIA小鼠的足肿胀、关节炎评分以及血清中促炎因子TNF-α的含量(P<0.01),组织病理学检查结果表明,各给药组改善了CIA小鼠的关节损伤,减少膝关节炎性细胞浸润,踝关节ATF-6、GRP78蛋白及mRNA的表达显著降低(P<0.01),CHOP蛋白及mRNA的表达显著升高(P<0.05)。结论:提示通过调控内质网应激通路降低RA小鼠炎症反应可能是金铁锁改善RA滑膜炎症的潜在机制。

Abstract:

Psammosilene tunicoides,a commonly used Yunnan folk medicine for the treatment of rheumatic paralysis, is used in modern research and treatment of RA. In vivo experiments were conducted based on the endoplasmic reticulum stress pathway to explore the mechanism of action of Psammosilene tunicoides extract (CEPT). Methods: Collagen-induced rheumatoid arthritis (CIA) mouse model was used, and the dosage of each administration group was continuously administered for 21 days. Foot swelling and arthritis scores, histopathological staining, and enzyme-linked immunosorbent assay (ELISA) were used to detect the tumor necrosis factor-α (TNF-α) content in mouse serum. Western blot method was used to determine the expression of activator of transcription factor 6 (ATF-6), glucose-regulated protein 78 (GRP78), and C/EBP homologous protein (CHOP) proteins in mouse ankle joints; real-time fluorescence quantitative nucleic acid amplification (real-time quantitative PCR, q-PCR detected the mRNA expression of ATF-6, GRP78, and CHOP in cells. Results: Each administration group significantly reduced foot swelling, arthritis scores, and serum pro-inflammatory factor TNF in CIA mice -α content (P<0.01), histopathological examination results showed that each administration group improved the joint damage of CIA mice, reduced knee arthritic cell infiltration, and the expression of ATF-6 and GRP78 protein and mRNA in the ankle joint Significantly reduced (P<0.01), CHOP protein and mRNA expression significantly increased (P<0.05). Conclusion: It is suggested that reducing the inflammatory response in RA mice by regulating the endoplasmic reticulum stress pathway may be the potential of Psammosilene tunicoides to improve RA synovial inflammation. mechanism.