Gui Ming, Li Yang, Wu Ting, Li Ying, Huang Ying, Zhang Hao
Department of Nephrology, Third Xiangya Hospital, Central South University, Changsha, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 Sep;28(9):915-9.
To determine the effects of glycyrrhetinic acid (GA), methotrexate(MTX) and GA+MTX on the expressions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in fibroblast-like synovial (FLS) cells from collagen induced arthritis (CIA) rats.
CIA-FLS cells were isolated and purified from the synovium of CIA rats by tissue block culture method, and then treated with MTX, GA, GA+MTX or nothing (control group), respectively. Each group was cultured for 3 different time periods (1, 3, 5 d). Real-time qPCR was used to detect the expressions of TNF-α and IL-1β mRNA in CIA-FLS cells and ELISA to determine the levels of TNF-α and IL-1β in the supernatant.
Day 1: No significant down-regulation of TNF-α and IL-1β mRNA was identified in intervention groups compared with the control group (P>0.05); Day 3: Intervention groups significantly down-regulated TNF-α and IL-1β mRNA (P<0.05 vs control), but without intergroup difference(P>0.05); Day 5: the three intervention groups all suppressed the expressions of TNF-α and IL-1β mRNA(P<0.05 vs control), with GA+MTX>MTX>GA in inhibiting effect (P<0.05). All intervention groups were found to inhibit the expressions of TNF-α and IL-1β in a time-dependent manner (P<0.05). Similar outcomes were observed in the change of the levels of TNF-α and IL-1β in cell culture supernatant by ELISA.
GA, MTX, GA+MTX suppress the expression of TNF-α and IL-1β in CIA-FLS cells in a time-dependent manner, and the suppressing effect is GA+MTX>MTX>GA.
确定甘草次酸（GA）、甲氨蝶呤（MTX）及GA+MTX对胶原诱导性关节炎（CIA）大鼠成纤维样滑膜（FLS）细胞中肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）表达的影响。
采用组织块培养法从CIA大鼠滑膜中分离并纯化CIA-FLS细胞，然后分别用MTX、GA、GA+MTX处理或不处理（对照组）。每组培养3个不同时间段（1、3、5天）。采用实时定量PCR检测CIA-FLS细胞中TNF-α和IL-1β mRNA的表达，并用ELISA法测定上清液中TNF-α和IL-1β的水平。
第1天：与对照组相比，各干预组TNF-α和IL-1β mRNA均无明显下调（P>0.05）；第3天：各干预组TNF-α和IL-1β mRNA明显下调（与对照组相比P<0.05），但组间无差异（P>0.05）；第5天：三个干预组均抑制TNF-α和IL-1β mRNA的表达（与对照组相比P<0.05），抑制作用为GA+MTX>MTX>GA（P<0.05）。所有干预组均呈时间依赖性抑制TNF-α和IL-1β的表达（P<0.05）。ELISA检测细胞培养上清液中TNF-α和IL-1β水平的变化也观察到类似结果。
GA、MTX、GA+MTX均呈时间依赖性抑制CIA-FLS细胞中TNF-α和IL-1β的表达，抑制作用为GA+MTX>MTX>GA。
