摘要
目的:探索转化生长因子-β1(TGF-β1)对大鼠肝星状细胞端粒酶表达的影响以及肝纤维化过程中TGF-β1所起的作用。方法:梯度密度离心法分离正常大鼠肝脏中原代肝星状细胞并分两组培养。肝星状细胞的形态通过荧光倒置显微镜观察,肝星状细胞的纯度通过免疫细胞化学染色和自发荧光鉴定。分别观察用浓度为0,0.1,1,10 ng/mL的TGF-β1处理24 h和1 ng/mL TGF-β1处理并培养3,6,9 d,观察肝星状细胞的端粒酶反转录酶(telomerase reverse transcriptase,TERT)mRNA的表达变化。结果:细胞形态学观察结果表明TGF-β1能触发HSCs从静息状态向高度分化的肌成纤维细胞的转化。正常大鼠原代肝星状细胞中TERT mRNA的表达随细胞培养时间的增加而增加,但相邻时间点间差异无统计学意义(P>0.05)。与0 ng/mL组相比,0.1 ng/mL TGF-β1处理HSCs 1 d对TERT mRNA的表达无明显抑制作用,而浓度为1 ng/mL和10 ng/mL TGF-β1处理HSCs 1 d能显著抑制TERT mRNA的表达(P<0.05)。用1 ng/mL TGF-β1处理HSCs 3 d对TERT mRNA的表达无明细抑制作用,1 ng/mL TGF-β1处理HSCs 6 d能显著抑制TERT mRNA的表达(P<0.05)。TGF-β1对HSCs中TERT mRNA的抑制作用呈浓度和时间依赖性。结论:肝纤维化过程中,TGF-β1可能通过减少端粒酶反转录酶的表达来促进肝星状细胞的转化,从而促进肝纤维化的进展。
Objective: To determine the ef ect of transforming growth factor-β1(TGF-β1) on the expression of telomerase in hepatic stellate cells(HSCs) in rats and the role of TGF-β1 in the development of liver i brosis. Methods: Primary HSCs were isolated from normal rats by density gradient separation and divided into 2 groups for culturing. h e morphology of HSCs was identii ed by the inverted l uorescence microscope. h e purity of HSCs was identii ed by immunohistological expression and l uorescence analysis. One group of HSCs was treated with dif erent concentrations(0, 0.1, 1, and 10 ng/mL) of TGF-β1 for 24 h, while the other group was treated with 1 ng/mL TGF-β1 and cultured for 3, 6, and 9 days. h e mRNA expression of telomerase reverse transcriptase(TERT) was assessed and compared by polymerase chain reaction. Results: Cell morphology showed that TGF-β1 triggered the differentiation of HSCs from a quiescent phenotype into highly activated myoi broblasts. TERT mRNA expression in the primary HSCs showed slight increase with the culture time, though with no statistical dif erence between the results at various time points(P>0.05). TGF-β1 at 0.1 ng/mL did not signii cantly af ect the TERT mRNA level compared with the 0 ng/mL group, while 1 ng/mL and 10 ng/mL TGF-β1 significantly decreased the level of TERT mRNA(P<0.05). TGF-β1 at 1 ng/mL had only weak effect on TERT mRNA expression after the 3 day treatment compared with the 0 ng/mL group(P>0.05). TGF-β1 at 1 ng/mL significantly inhibited TERT mRNA expression 6 days after the treatment(P<0.05). TGF-β1 inhibited the expression of TERT mRNA level in the HSCs in both dose- and time-dependent manner. Conclusion: TGF-β1 may contribute to the transdif erentiation of HSCs by reducing TERT levels to develop hepatic i brosis.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2014年第5期442-451,共10页
Journal of Central South University ：Medical Science
关键词
转化生长因子-Β1
肝星状细胞
端粒酶反转录酶
肝纤维化
transforming growth factor-β1
hepatic stellate cell
telomerase reverse transcriptase
hepatic fibrosis
