水溶性乌梢蛇总蛋白对成纤维样滑膜细胞分泌白介素-1β、α-肿瘤坏死因子和白介素-10的影响

Effect of soluble total proteins of Zaocys dumnades on interleukin-1β,tumor necrosis factor-α and interleukin-10 expressions in human fibroblast-like synoviocytes in vitro

目的观察水溶性乌梢蛇总蛋白对类风湿关节炎(RA)成纤维样滑膜细胞分泌IL-1β、TNF-α和IL-10的影响,以探讨治疗RA的可能机制。方法用常规方法提取水溶性乌梢蛇总蛋白,取RA患者滑膜组织进行成纤维样滑膜细胞培养;选择对数生长期的细胞进行实验,实验分五组,分别加入昆明山海棠(200μg/ml,阳性对照组)、水溶性乌梢蛇总蛋白(50、150、450μg/ml)DMEM溶液,及加入等体积的DMEM溶液(空白对照组)进行细胞培养,72小后采用ELISA方法测定培养液上清中IL-1β、TNF-α和IL-10的水平,RT-PCR检测滑膜细胞IL-1β、TNF-α和IL-10的mRNA表达。结果水溶性乌梢蛇总蛋白150μg/ml、450μg/ml组和阳性对照组细胞培养液上清中TNF-α、IL-1β水平明显降低,mRNA表达减少,而IL-10水平升高,mRNA表达增加,与空白对照组差异具有统计学意义(P<0.05)。结论水溶性乌梢蛇总蛋白可能通过抑制成纤维样滑膜细胞TNF-α和IL-1β及促进IL-10分泌来减轻炎症反应,到达治疗RA的目的。

Objective To evaluate the effect of soluble total proteins of Zaocys dhumnades on the expressions of interleukin-1β（IL-1β）, tumor necrosis factor-ct （TNF-α） and IL-10 in human fibroblast-like synoviocytes （FLS） cultured in vitro. Methods Primary cultured FLS isolated fi＇om the synovium of patients with rheumatoid arthritis （RA） were incubated in the presence of different concentrations （50, 150 and 450 μg/ml） of soluble total proteins of Zaocys dhumnades, with Tripterygiurn hypoglaucurn Hutch （THH） and DMEM as the positive and negative controls, respectively. Enzyme-linked immunosorbent assay （ELISA） and RT-PCR were used to detect the expressions oflL-1β, IL-10 and TNF-α in the FLS. Results The protein and mRNA levels of IL-1β and TNF-α in the supernatant of the FLS exposed to 150 and 450 μg/ml of the soluble total proteins ofZaocys dhurnnades decreased, while IL-10 protein and mRNA increased significantly as compared with those in the negative control group （P〈0.01）. Conclusion The soluble total proteins ofZaocys dhurnnades produce therapeutic effect on RA possibly by inhibiting IL- 1β and TNF-α and promoting IL- 10 expressions in the FLS.