TGF-β1介导新生大鼠心肌成纤维细胞Ⅰ型和Ⅳ型胶原合成的研究(英文)

目的:观察TGF-β1对原代培养的新生大鼠心肌成型与Ⅳ胶原表达的影响。方法:取出生3~7d的SD大鼠心脏,利用酶消化法结合差速贴壁体外培养心肌成纤维细胞;分别用不同浓度的TGF-β1(TGF-β1=10 ng/mL、1ng/mL、0 ng/mL)干预心肌成纤维细胞,分别于干预后5h、24h,采用RT-PCR检测I型胶原和Ⅳ型胶原的表达。结果:1.用0ng/mL(对照组),1ng/mL和10ng/mL的TGF-β1干预心肌成纤维细胞5h,RT-PCR检测I型胶原/β-actin光密度值分别为0.61±0.02,0.73±0.03和0.86±0.04,光密度值随着TGF-β1浓度的增加而增加,各组比较,差异有显著性(P<0.01);干预心肌成纤维细胞24h后,RT-PCR检测I型胶原/β-actin光密度值分别为0.65±.03,0.91±0.02和0.98±0.02,光密度值随着TGF-β1浓度的增加而增加,各组比较,差异有显著性(P<0.01)。2.用0 ng/mL(对照组),1ng/mL and 10ng/mL的TGF-β1干预心肌成纤维细胞5h,RT-PCR检测Ⅳ型胶原/β-actin光密度值分别为0.58±0.06,0.71±0.02,0.87±0.02,光密度值随着TGF-β1浓度的增加而增加,各组比较,差异有显著性(P<0.01);干预心肌成纤维细胞24h后,RT-PCR检测Ⅳ型胶原/β-actin光密度值分别为0.62±0.01,0.83±0.05,0.96±0.02,光密度值随着TGFβ1浓度的增加而增加,各组比较,差异有显著性(P<0.01)。结论:TGFβ1能加强I型和Ⅳ型胶原的表达,在24小时内呈时间和剂量依赖性。

TGF-1β Mediates Collagen Ⅰ and Collagen Ⅳ Synthesis in Neonate Rat Cultured Cardiac Fibroblasts

Objective To investigate the effect of TGF-β1 on synthesis of collagen I and collagen Ⅳ.Methods The cardiac fibroblasts wereisolated from SD mouse, which were treated using different concentrations of TGF-β1 ie.1ng/mL and 10ng/mL, then incubated for a specific duration of time.Cells were divided into three main groups(TGF-β1 10ng/mL, 1ng/mL and 0ng/mL).As stated above that all groups were observed for a given time(5 hrs or 24 hours).Then RT-PCR were used to detect biosynthesis of collagen type I and collagen Ⅳ respectively.Results 1.The primary cultured rat myocardial fibroblasts were treated using different concentrations of TGF-β1(0ng/mL, for control group; 1ng/mL and, and 10ng/mL for test groups respectively)after 5 hrs the OD values of collagen I/β-actin checked with RT-PCR were 0.61±0.02, 0.73±0.03 and 0.86±0.04 respectively, and after 24 h the OD values of collagen I/β-actin for control group, 1ng/ml and 10ng/mL were 0.65±0.03, 0.91±0.02 and 0.98±0.02 respectively(all P＜0.01 compared with controls, and other groups).And treated the cells with the same condition, the OD values for collagen Ⅳ/β-actin were 0.58±0.06, 0.71±0.02, 0.87±0.02 respectively, 24h the OD values of RT-PCR were 0.62±0.01, 0.83±0.05, 0.96±0.02(all P＜0.01 compared with controls, and other groups)respectively.Conclusion TGF-β1 can enhance biosynthesis of collagen I and Ⅳ in a dose and time dependent manner during 24 hours observation period.