Wang Yan, Liu Xin-min
Department of Geriatrics, Peking University First Hospital, Beijing100034, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2006 Oct 18;38(5):506-9.
To investigate the tendency and correlation between connective tissue growth factor (CTGF) and low density lipoprotein receptor-related protein (LRP) in human pulmonary fibroblasts-1 (HPF-1) induced by transforming growth factor beta1 (TGF-beta1) at different times.
After the HPF-1 cells were stimulated with TGF-beta1 (5 microg/L) at different times (0 h, 3 h, 6 h,12 h and 24 h),CTGF and LRP mRNA expressions were analyzed by RT-PCR. The same preparation steps of cell culture were repeated, then, the protein expressions were determined by Western Blot, co-immunoprecipitation Western Blot and immunocytochemistry.
The expressions of CTGF and LRP mRNA had similar tendency, and LRP (224.87 +/- 7.00) correlated with CTGF (131.53 +/- 2.86) positively (r = 0.8402, P < 0.05). Furthermore, the protein expression levels had significant difference between 0 h (190.85 +/- 2.86) and 3 h (222.45 +/- 3.66) groups (F = 18.06, P < 0.01),and by immunoprecipitation, it was discovered that the quantity of CTGF bounded to LRP changes, which was similar to the result of LRP western blot. The analysis of immunocytochemistry disclosed the like results as well (3 h: 88.66 +/- 15.72, 0 h: 27.56 +/- 6.72, F = 244.36, P < 0.01).
The expression of LRP coincides with that of CTGF at the same stimulated time point of TGF-beta1. It suggests that LRP may be involved in the biological process of CTGF. And further research about LRP may provide a new direction to pulmonary fibrosis therapy.
探讨转化生长因子β1(TGF-β1)在不同时间诱导人肺成纤维细胞-1(HPF-1)中结缔组织生长因子(CTGF)与低密度脂蛋白受体相关蛋白(LRP)之间的变化趋势及相关性。
用TGF-β1(5μg/L)在不同时间点(0 h、3 h、6 h、12 h和24 h)刺激HPF-1细胞后,采用逆转录-聚合酶链反应(RT-PCR)分析CTGF和LRP mRNA表达。重复相同的细胞培养制备步骤,然后用蛋白质印迹法、免疫共沉淀蛋白质印迹法和免疫细胞化学法检测蛋白质表达。
CTGF和LRP mRNA表达具有相似趋势,且LRP(224.87±7.00)与CTGF(131.53±2.86)呈正相关(r = 0.8402,P < 0.05)。此外,0 h组(190.85±2.86)与3 h组(222.45±3.66)蛋白质表达水平差异有统计学意义(F = 18.06,P < 0.01),通过免疫沉淀发现与LRP结合的CTGF量发生变化,这与LRP蛋白质印迹结果相似。免疫细胞化学分析也显示了类似结果(3 h:88.66±15.72,0 h:27.56±6.72,F = 244.36,P < 0.01)。
在TGF-β1相同刺激时间点,LRP与CTGF表达一致。提示LRP可能参与CTGF的生物学过程。对LRP的进一步研究可能为肺纤维化治疗提供新方向。