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目的:研究白细胞介素-1受体拮抗剂(IL-1ra)对大鼠肝星状细胞(HSC)Ⅰ型胶原(Col-Ⅰ)、Ⅲ型胶原(Col-Ⅲ)和基质金属蛋白酶组织抑制因子1(TIMP-1)基因表达的影响,探讨IL-1ra抗肝纤维化作用的机制。方法:原位分离、培养大鼠HSC,荧光显微镜观察及免疫细胞化学染色法鉴定。用1μg/L、10μg/L、100μg/L3个浓度的IL-1ra分别作用于HSC48h,实时荧光定量RT-PCR法检测Col-Ⅰ、Col-Ⅲ和TIMP-1mRNA的表达。结果:对照组Col-Ⅰ、Col-Ⅲ和TIMP-1mRNA的表达水平分别为1.97±0.29、2.70±0.57、3.83±0.30。加入1μg/L、10μg/L、100μg/LIL-1ra后Col-Ⅰ基因表达水平依次为1.34±0.35、0.86±0.19、0.35±0.03;Col-Ⅲ基因表达水平依次为2.02±0.29、1.13±0.09、0.47±0.11;TIMP-1mRNA的表达水平依次为3.12±0.25、2.53±0.38、1.98±0.18,与对照组比较,不同浓度处理组Col-Ⅰ、Col-Ⅲ和TIMP-1mRNA的表达均明显降低(P<0.05)。结论:IL-1ra以剂量依赖方式抑制HSC中Col-Ⅰ、Col-Ⅲ及TIMP-1mRNA的表达,具有抗纤维化作用。
OBJECTIVE: To study the effects of interleukin-1 receptor antagonist (IL-1ra) on the tissue of type Ⅰ collagen, type Ⅲ collagen and matrix metalloproteinase in rat hepatic stellate cells (HSC) 1 (TIMP-1) gene expression, explore the mechanism of IL-1ra anti-liver fibrosis. Methods: HSCs were isolated and cultured in situ. The HSCs were observed by fluorescence microscopy and immunocytochemical staining. The expression of Col-Ⅰ, Col-Ⅲ and TIMP-1 mRNA was detected by real-time fluorescence quantitative RT-PCR after treated with 1μg / L, 10μg / L and 100μg / Results: The expression of Col-Ⅰ, Col-Ⅲ and TIMP-1 mRNA in the control group were 1.97 ± 0.29, 2.70 ± 0.57 and 3.83 ± 0.30, respectively. The gene expression of Col-Ⅰ was 1.34 ± 0.35,0.86 ± 0.19,0.35 ± 0.03 after added with 1μg / L, 10μg / L and 100μg / L IL-1ra respectively. The gene expression of Col-Ⅲ was 2.02 ± 0.29,1.13 ± 0.09 , 0.47 ± 0.11 respectively. The expression of TIMP-1 mRNA was 3.12 ± 0.25,2.53 ± 0.38,1.98 ± 0.18 in turn, compared with the control group, the expression of Col-Ⅰ, Col-Ⅲ and TIMP-1 mRNA were significantly decreased (P <0.05). CONCLUSION: IL-1ra inhibits the expression of Col-Ⅰ, Col-Ⅲ and TIMP-1 mRNA in HSC in a dose-dependent manner and has anti-fibrosis effects.