BCAT1对HSC-LX2人肝星状细胞纤维化相关基因表达的影响

The effect of BCAT1 on the expression of fibrosis related genes in HSC-LX2 human stellate cells

目的探讨支链氨基酸转氨酶1(BCAT1)对HSC-LX2人肝星状细胞纤维化相关基因表达的影响。方法构建携带BCAT1基因的慢病毒载体,将病毒转染HSC-LX2细胞,运用q RT-PCR和Western blotting检测转染BCAT1过表达病毒后的HSC-LX2细胞中BCAT1、ACTA2、TIMP1、MMP2和COL1A1基因的表达情况。结果携带BCAT1基因的慢病毒转染HSC-LX2细胞后,实验组(转染p-BCAT1)HSC-LX2细胞和对照组(转染Vec)HSC-LX2细胞相比,促进纤维化的ACTA2、TIMP1、COL1A1基因的m RNA及蛋白表达均升高,抗纤维化的MMP2基因的m RNA及蛋白表达均下降,两者的差异均具有统计学意义(P<0.05)。结论 BCAT1增强HSC-LX2人肝星状细胞中ACTA2、TIMP1、COL1A1促纤维化相关基因的表达,抑制抗纤维化相关基因MMP2的表达。

objective To investigate the effect of branched-chain aminotransferases 1（BCAT1） on expression of fibrosis-related genes in HSC-LX2 human hepatic stellate cells. Methods The lentiviral vector expressing BCAT1 gene was constructed and transduced into HSC-LX2 cells. After the exogenous expression of BCAT1 in HSC-LX2 cells, the m RNA and protein expression levels of ACTA2, TIMP1, MMP2 and COL1 A1 were detected by q RT-PCR and Western blotting. Results After BCAT1 transduction, the HSC-LX2 cells in the p-BCAT1 group were compared with those in Vec group. We found that the m RNA and protein levels of ACTA2, TIMP1 and COL1 A1 were up-regulated while the m RNA and protein expression levels of MMP2 were down-regulated in p-BCAT1 group. The differences were both statistically significant（P〈0.05）. Conclusion BCAT1 enhances the expression levels of ACTA2, TIMP1 and COL1 A1 in HSC-LX2 cells which accelerate fibrosis, and inhibits the expression levels of MMP2 which restrain fibrosis.