摘要
目的观察干扰素γ(IFN γ)对肝星状细胞(HSC)-T6细胞Ⅰ型胶原(ColⅠ)、Ⅲ型胶原(colⅢ)、基质金属蛋白酶组织抑制因子1(TIMP1)基因表达的影响。方法浓度0.1、1、10、102、103、104、2.5×105、5×105U/ml IFN γ作用于HSC-T6细胞48 h,采用四甲基偶氮唑盐比色法观察其对HSC-T6细胞生长指数的影响;以1、102、104U/ml IFN γ作用于HSC-T6细胞48 h,采用逆转录聚合酶链反应方法测定其对HSC-T6细胞Col Ⅰ、ColⅢ、TIMP1基因表达的影响。结果空白对照组ColⅠ、ColⅢ、TIMP1基因表达水平分别为2.86±0.21、2.00±0.23、3.90±0.81;1U/m1、102U/ml、104U/ml IFNγ作用于HSC-T6细胞,ColⅠ基因表达水平依次为1.00±0.11、0.42±0.12、0.33±0.12,与空白对照组比较, 差异有统计学意义;ColⅢ基因表达水平依次为1.09±0.1 5、0.52±0.14、0.43±0.18,与空白对照组比较, 差异有统计学意义;TIMP1基因表达水平依次为3.81±0.37、3.50±0.51、3.41±0.31,与空白对照组比较,差异无统计学意义。结论IFNγ明显抑制HSC-T6细胞colⅠ、ColⅢ基因表达水平,而对TIMP1基因表达水平无明显影响,从而可抑制Ⅰ、Ⅲ型胶原的合成,这可能是其抗纤维化的作用机制之一。
Objective To observe the in vitro effects of gamma-interferon (IFN γ ) on gene expression of collagen Ⅰ (Col Ⅰ), Ⅲ (Col Ⅲ) and tissue inhibitor of metalloprotenase 1 (TIMP1) of HSC-T6 cells. Methods Cultured HSC-T6 cells were exposed to IFN γ at concentrations of 0.1, 1, 10, 102, 103, 104, 2.5×105, 5×105 U/ml for 48 hours. 4,5-simethylthiazaoly colormetric assay was used to evaluate the effect of IFN γ on HSC-T6 cell proliferation. After incubating with IFN γ (1 U/ml, 102 U/ml and 104 U/ml) for 48 hours, HSC-T6 cells were harvested to detect Col Ⅰ, Col Ⅲ and TIMP1 steady state mRNA levels by quantitative reverse-transcription polymerase chain reaction (RT-PCR). Results The Col Ⅰ, Col Ⅲ and TIMP1 mRNA levels of the control group were 2.86±0.21, 2.00±0.23 and 3.90±0.81, respectively. Col Ⅰ and Col Ⅲ mRNA levels in HSC-T6 cells treated by different concentrations of IFN γ were lower than that of the controls (P < 0.01). There was no significant difference in TIMP1 mRNA levels between IFN γ groups and controls. Conclusion IFN γ suppresses expression of Col Ⅰ and Col Ⅲ whereas it has no effect on TIMP1 mRNA expression. The antifibrotic mechanism of IFN γ may be partly due to its down-regulation of Col Ⅰ1 and Col Ⅲ mRNA levels in HSC-T6 cells.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2005年第7期528-530,共3页
Chinese Journal of Hepatology
基金
北京市科技新星计划(954810500)