中文摘要：

目的探讨胰岛素样生长因子结合蛋白相关蛋白l（IGFBPrPl）对肝星状细胞转化生长因子β1（TGF-β1）／Smad3信号通路的影响及意义。方法体外培养大鼠肝星状细胞株HSC-T6，分别设立空白对照组（加入等量磷酸盐缓冲液）和IGFBPrPl处理组（加入终浓度为30μg/L的IGFBPrPl），作用24h后收集细胞培养上清液，提取全细胞蛋白，另外收集细胞爬片。培养上清液中TGF-β1的表达采用Western印迹法及ELISA法检测，HSC-T6细胞TGF-β1的表达采用免疫细胞化学法检测；培养上清液中Smad3、p-Smad2／3和纤维连接蛋白（Fn）的表达采用Western印迹法检测。空白对照组与IGFBPrPl处理组TGF-β1、Smad3、p-Smad2／3、Fn蛋白表达量、p-Smad2／3与Smad3蛋白相对表达量比值比较采用独立样本t检验。结果Western印迹法及ELISA检测结果均显示IGFBPrPl处理组培养上清液TGF-β1蛋白表达量均明显高于空白对照组[0．34±0．06与0．19±0．03相比，（188．34±32．24）ng／L与（136．16±19．52）ng／L相比]，免疫细胞化学法也显示HSC-T6细胞TGF-β1蛋白表达量明显高于空白对照组（11．89±0．32与7．71±0．63相比），且差异均有统计学意义（t=3．859，P=0．018；t=3．391，P〈0．05；t=14．490，P〈0．05）；IGFBPrPl处理组与空白对照组HSC-T6细胞中Smad3相对表达量差异无统计学意义（0．65±0．05与0．64±0．05相比，t=0．084，P=0．937），p-Smad2／3蛋白相对表达量、p-Smad2／3与Smad3蛋白相对表达量的比值均明显高于空白对照组，且差异均有统计学意义（0．70±0．05与0．46±0．03相比，t=8．050，P=0．001；1．10±0．12与0．72±0．05相比，t=5．181，P=0．007）；IGFBPrPl处理组培养上清液中Fn蛋白相对表达量明显高于空白对照组，且差异有统计学意义（0．52±0．04与0．35±0．02相比，t=5．976，P=0．004）。结论IGFBPrPl可促进体外培养的HSC-T6产生TGF-β1。IGFBPrPl?

英文摘要：

Objective To investigate the effect of insulin-like growth factor binding protein related protein 1 （IGFBPrP1） on transforming growth factor [31 （TGF-β1）/Smad3 signaling pathway in hepatic stellate cells. Methods HSC-T6 cells were cultured in vitro and divided into control group （ incubated with equal phosphate buffer saline） and IGFBPrP1 group （ treated with 30 μg/L of IGFBPrP1 ） , then the culture supernatant fluid was collected and whole-cell proteins were extracted and the slides of cells were attained after 24 hours. The expression of TGF-β1 in culture supernatant fluid was detected by both Western blot and ELISA and the expression of TGF-β1in HSC-T6 cells was detected by immunocytochemieal staining; the expressions of Smad3, p-Smad2/3 and fibronectin （Fn） in culture supernatant fluid were detected by Western blot. Comparison between two groups was done with independent sample t-test. Results The results of Western blot and ELISA showed that the expression of TGF-I31 in culture supernatant fluid in IGFBPrP1 group was significantly higher than that in control group [ （0.34 ± 0.06 ） vs （0.19± 0. 03 ）, t = 3. 859, P = 0.018：（188.34 ±32.24） ng/L vs （136. 16± 19.52） ng/L, t=3. 391. P〈0.057. The result of immunocytochemical staining also showed that the expression of TGF-β1 in HSC-T6 cells in IGFBPrPI group was significantly higher than that in control group [ （ 11.89 ± 0.32） vs （7.71 ± 0.63）, t = 14. 490, P 〈 0.05 ]. The difference of the expression of Smad3 in HSC-T6 cells between IGFBPrP1 group and control group was not statistically significant [ （0.65 ± 0.05 ） vs （0.64 ± 0.05 ）, t = 0. 084, P = 0. 937 ]. The expression of p-Smad2/3 and the ratio of p-Smad2/3 and Smad3 in IGFBPrP1 group were higher than those in control group, and the differences were statistically significant [ （ 0.70 ± 0. 05 ） vs （ 0. 46 ± 0. 03 ）, t =8. 050, P=0.001; （1.10＋0.12） vs （0.72＋0.05）, t=5.181, P=0.007]. The expression of Fnin culture supe