京尼平苷酸对佐剂性关节炎模型大鼠滑膜细胞体外培养增殖及分泌细胞因子的影响

目的:研究京尼平苷酸(GA)对佐剂性关节炎(AA)模型大鼠滑膜细胞体外培养增殖能力和分泌细胞因子的影响。方法:用弗氏完全佐剂建立AA模型大鼠,分离其滑膜细胞进行培养,以阳性细胞数和纯度鉴定其免疫细胞;将细胞分为对照组,GA低、中、高(10-7、10-6、10-5mol·L-1)剂量组,阳性对照(甲氨蝶呤,10-6mol·L-1)组并加入相应药物处理。MTT法检测吸光度(A)考察GA对滑膜细胞增殖的影响;流式细胞术测定细胞周期;酶联免疫吸附法检测各组滑膜细胞分泌肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-10的水平。结果:2～3代滑膜细胞纯度达到试验要求;与对照组比较,GA高剂量组和阳性对照组大鼠滑膜细胞A值,S和G2/M期细胞比例,细胞外液TNF-α、IL-1β水平明显降低(P<0.05或P<0.01),G1期细胞比例、IL-10水平明显升高(P<0.05或P<0.01)。结论:GA能显著抑制AA模型大鼠滑膜细胞的增殖,阻滞细胞周期于G1期,抑制滑膜细胞分泌TNF-α和IL-1β,促进IL-10分泌。

Effect of Geniposide Acid on the Proliferation and Cytokines Secretion of Synoviocytes in Rats with Adjuvant-induced Arthritis in Vitro

OBJECTIVE： To study the effect of geniposide acid （GA） on the proliferation and cytokines secretion of synoviocytes in rats with adjuvant-induced arthritis （AA） in vitro. METHODS： AA rats were prepared using FCA, and synoviocytes were isolated and cultured. The amount and purity of positive cell were used to indentify immune cell. Synoviocytes were divided into control group, GA low-dose group （10-7 mol·L^-1）, GA medium-dose group （10-6 mol·L^-1） and GA high-dose group （10-5 mol·L^-1）, positive control group （methotrexate, 10-6 mol·L^-1）. MTT assay was used to determine absorbance （A） of synoviocytes to investigate the effect of GA on the proliferation of synoviocytes. Cell cycle was analyzed by flow cytometry. The level of TNF-α and IL-1β and IL-10 were determined by enzyme linked immunosorbent assay （ELISA）. RESULTS： The purity of 2～3 generation of synoviocytes was up to the experimental demand. Compared with control group, the value of A, the proportion of S phase cell and G2/M phase cell, the levels of TNF-α and IL-1β in extracellular fluid in GA high-dose group and positive control group were decreased significantly （P〈0.05 or P〈0.01）; the proportion of G1 phase cell and IL-10 were increased significantly （P〈0.05 or P〈0.01）. CONCLUSION： GA can significantly inhibit the proliferation of synoviocytes in AA rat, block cell cycle progression in G1 phase, inhibit the secretion of TNF-α and IL-1β in synoviocytes, but promote secretion of IL-10.