The effect of TGF-β1 on cell proliferation,cell cycle regulation and collagen expression in HSC-T6 cells
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国家科技期刊平台
NETL
NSTL
维普
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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% vs 26.4±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COL ⅢmRNA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.
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-myccdk-2,cyclinE,EGF, HGF, Bel-2,NF-kB, MMP1, MMP9, MMP14, TIMP-1, PAI-l,a-SMA, Collagen-I and Collagen-M genes; Collagen-I, Collagen-M ,and a-SMA secreted by HSC-T6 cells were detected by ELISA. Results In comparison to HSC-T6 cells in control group,TGF-J31 decreased the cell counts in G0-Gi 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-(31 treatment increased the mRNA levels of SMAD3, c-myc, cdk2, cyclin E, EGF, Bcl-2, NF-kB, TIMP1,PAI-1, a-SMA, and Collagn-I after treatment for 24 and 36 hours. Conversely, the mRNA level of HGF, MMPl,MMP9,MMP14,and collagen-III decreased at 24 hr,but increased at 36 hr. TGF -pi 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 a-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.
国家科技期刊平台
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维普
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