中文摘要：

目的观察TGF-β1对大鼠肝星状细胞系HSC-T6增殖、细胞周期以及分泌肝纤维化相关胶原的影响。方法采用MTT法检测细胞增殖;使用流式细胞仪检测细胞周期;采用荧光定量RT-PCR法检测SMAD3、c-myc、cdk-2、cyclinE、EGF、HGF、Bcl-2、NF-κB、MMP1、MMP9、MMP14、TIMP-1、PAI-1、α-SMA、COLⅠ及COLⅢ等基因mRNA水平;采用ELISA法检测细胞培养液COLⅠ、COLⅢ和α-SMA含量。结果与对照组比,TGF-β1处理24h及36h时,细胞形态及生长状况均无明显变化;在24h和36h时,TGF-β1处理组细胞G0/G1期细胞比率均减少（24h：57.3±8.5%vs 60.6±9.7%;36h：53.0±2.2%vs 56.6±5.0%）,S期细胞比例略高（24h：30.6±7.2%vs26.4±10.1%;36h：35.2±3.7%vs 30.8±2.5%）,但差异无统计学意义（P〉0.05）;TGF-β1处理组SMAD3、c-myc、cdk2、cyclin E、EGF、Bcl-2、NF-κB、TIMP1、PAI-1、α-SMA及COLⅠmRNA在24h和36h表达均上调,HGF、MMP1、MMP9、MMP14及COLⅢmRNA在24h时表达下调,36h时表达上调;TGF-β1处理组HSC-T6细胞分泌COLⅠ[24h：（63.0±7.4ng/ml vs 33.2±10.8 ng/ml,P〈0.05;36h：58.5±6.0ng/ml vs 42.2±6.3ng/ml,P〈0.05];α-SMA[24h：20.6±2.6ng/ml vs 4.2±0.7ng/ml,P〈0.05;36h：59.7±14.6ng/ml vs 36.8±5.6ng/ml,P〈0.05）均显著增加。结论 TGF-β1对大鼠肝星状细胞系HSC-T6显示了促增殖作用,并能促进肝纤维化相关胶原的分泌。

英文摘要：

Objective To explore the effect of TGF-β1 on cell proliferation,cell cycle regulation and collagen expression by HSC-T6 cell line.Methods The HSC-T6 cell proliferation was detected by MTT at 24 and 36 hours,respectively;The HSC-T6 cells were treated with TGF-β1（10 ng/ml） or not as control group.Flow cytometry was performed to measure the cell cycle;Fluorescent quantitative real time RT-PCR was used to quantify the mRNA levels of SMAD3,c-myc,cdk-2,cyclinE,EGF,HGF,Bcl-2,NF-κB,MMP1,MMP9,MMP14,TIMP-1,PAI-1,α-SMA,Collagen-I and Collagen-Ⅲ genes;Collagen-I,Collagen-Ⅲ,and α-SMA secreted by HSC-T6 cells were detected by ELISA.Results In comparison to HSC-T6 cells in control group,TGF-β1 decreased the cell counts in G0-G1 phase（24h： 57.3±8.5% vs 60.6±9.7%;36h： 53.0±2.2% vs 56.6±5.0%,both P〉0.05）,while it increased the cells in S phase（24h：30.6±7.2% vs 26.4±10.1%;36h：35.2±3.7% vs 30.8±2.5%,both P〉0.05）;We found that TGF-β1 treatment increased the mRNA levels of SMAD3,c-myc,cdk2,cyclin E,EGF,Bcl-2,NF-κB,TIMP1,PAI-1,α-SMA,and Collagn-I after treatment for 24 and 36 hours.Conversely,the mRNA level of HGF,MMP1,MMP9,MMP14,and collagen-III decreased at 24 hr,but increased at 36 hr.TGF-β1 promoted the secretion of collagen-I [24h：63.0±7.4ng/ml vs.33.2±10.8ng/ml,P〈0.05;36h：58.5±6.0ng/ml vs.42.2±6.3ng/ml,P〈0.05],and α-SMA[24h： 20.6±2.6ng/ml vs 4.2±0.7ng/ml,P〈0.05;36h：59.7±14.6ng/ml vs 36.8±5.6ng/ml,P〈0.05] by HSC-T6 cells after the treatment.Conclusion TGF-β1 can promote cell proliferation and the secretion of collagen of rat HSC-T6 cell lines.