胰岛素样生长因子结合蛋白7在活化肝星状细胞促肝纤维化中的作用  

Role of insulin-like growth factor binding protein 7 in the process of hepatic fibrosis promoted by activated hepatic stellate cells

作　　者：张骞骞[1] 刘立新[1] 郭晓红[1] 刘毅[1] 张东梅[1] 

机构地区：[1]山西医科大学第一医院科研实验中心、肝病研究所、细胞生理学省部共建教育部重点实验室,太原030001

出　　处：《中华消化外科杂志》2009年第6期457-459,共3页Chinese Journal of Digestive Surgery

基　　金：国家自然科学基金（30740031）;教育部新世纪优秀人才支持计划（NCET-06-0264）

摘　　要：目的探讨胰岛素样生长因子结合蛋白7（IGFBP7）对肝星状细胞-T6合成分泌纤维连接蛋白的调节作用及抗IGFBP7抗体诱导肝星状细胞-T6凋亡的作用。方法体外培养活化的肝星状细胞-T6分为5组：（1）空白对照组；（2）IGFBP720μg／L组；（3）抗IGFBP7抗体0．25mg／L组；（4）抗IGFBP7抗体0．50mg／L组；（5）抗IGFBP7抗体1．00mg／L组。采用Western blot及ELISA法检测肝星状细胞-T6经IGFBP7作用24h后纤维连接蛋白的表达及合成分泌量的变化；MTT比色法检测抗IGFBP7抗体对肝星状细胞-T6作用14h后增殖抑制的影响，同时用流式细胞仪测定抗IGFBP7抗体对肝星状细胞-T6凋亡发生率的影响。采用单因素方差分析，两样本均数比较采用t检验，多重比较采用SNK-q检验。结果肝星状细胞-T6经IGFBP7作用后纤维连接蛋白的表达及合成分泌量较空白对照组显著增加（t=22．06，7．43，P〈0．05）。抗IGFBP7抗体对肝星状细胞-T6的增殖具有明显抑制作用（F=14．70，P〈0．05）。抗IGFBP7抗体能够上调肝星状细胞-T6的凋亡率（F=63．79，P〈0．05）。结论IGFBP7能够使细胞外基质的重要组成成分纤维连接蛋白的合成与分泌增加，抗IGFBP7抗体可以抑制肝星状细胞-T6的增殖并诱导其发生凋亡。Objective To investigate the regulatory effect of insulin-like growth factor binding protein 7 （ IGFBP7 ） on the synthesis and secretion of fibronection and the inhibiting effect of anti-IGFBP7 antibody on apoptosis of hepatic stellate cell-T6 （HSC-T6）. Methods HSC-T6 cultured in vitro were divided into blank control group, IGFBP7 20μg/L group, anti-IGFBP7 antibody 0.25 mg/L group, anti-IGFBP7 antibody 0.50 mg/L group and anti-IGFBP7 antibody 1.00 mg/L group. HSC-T6 were treated by IGFBP7 for 24 hours, then the expression of fibronectin was detected by Western blot and the content of fibronectin by ELISA. After incubating HSC-T6 with anti-IGFBP7 antibody for 14 hours, the inhibiting effect of anti-IGFBP7 antibody on HST-T6 was evaluated by MTT assay, and the effect of anti-IGFBP7 on the apoptosis of HST-T6 was detected by flow cytometry. All data were analyzed by one-way ANOVA, t test and SNK-q test. Results The content and expression of fibronection in IGFBP7 20 mg/L group were significatttly higher than those in the blank control group （ t = 22, 06, 7, 43, P 〈 0.05 ）. Anti-IGFBP7 antibody had an obvious inhibiting effect on the proliferation of HSC-T6 （ F = 14.70, P 〈 0.05）, and it also enhanced the ratio of apoptosis of HSC-T6 （ F = 63.79, P 〈 0.05 ）. Conclusions IGFBP7 can increase the synthesis and secretion of fibronectin which is the principal component of extracellular matrix. Anti-IGFBP7 antibody can inhibit the proliferation and promote the apoptosis of HSC-T6.

关 键 词：胰岛素样生长因子结合蛋白7 肝星状细胞 纤维连接蛋白 凋亡 

分 类 号：R575.2[医药卫生—消化系统] R720.597[医药卫生—内科学]