PAP-M6P与RGDY对肝星状细胞活化功能的联合干预作用  

Combined Interventional Effects of PAP-M6P and RGDY on Activated Function of Hepatic Stellate Cells

作　　者：周韵斓[1] 吴建新[1] 熊敏莉[1] 付杰[2] 李定国[1] 张剑波[2] 

机构地区：[1]上海交通大学医学院附属新华医院消化内科,200092 [2]华东师范大学化学系

出　　处：《胃肠病学》2008年第11期651-654,共4页Chinese Journal of Gastroenterology

基　　金：国家自然科学基金(No.30672675)资助

摘　　要：背景:活化的肝星状细胞(HSC)表面6-磷酸甘露糖/胰岛素样生长因子Ⅱ(M6P/IGFⅡ)受体和整合素受体显著增加。外源性含有M6P或精氨酰-甘氨酰-天冬氨酸(RGD)基团的分子可干预HSC活化,但目前尚缺乏两者比较或联合应用的研究。目的:探讨人工化学合成的p-氨基苯基-6-磷酸-α-D-甘露糖(PAP-M6P)和精氨酰-甘氨酰-天冬氨酰-酪氨酸(RGDY)小肽对HSC活化功能的联合干预作用。方法:取培养10d的活化期HSC,分别设空白对照组、RGDY对照组、PAP-M6P低、中、高浓度组以及低、中、高浓度联合组进行干预。以甲基噻唑基四唑(MTT)法检测细胞抑制率,以逆转录聚合酶链反应(RT-PCR)检测转化生长因子(TGF)-β1mRNA表达,以酶联免疫吸附测定(ELISA)检测Ⅰ型胶原、透明质酸和活化的TGF-β1蛋白含量。结果:各组细胞抑制率无明显差异。中浓度联合组TGF-β1mRNA表达和中、高浓度联合组活化的TGF-β1蛋白含量显著低于RGDY对照组和相应浓度PAP-M6P组(P<0.05);高浓度联合组Ⅰ型胶原、透明质酸含量显著低于RGDY对照组和各浓度PAP-M6P组(P<0.05)。结论:PAP-M6P与RGDY联合应用能显著抑制HSC表达和激活TGF-β1,并减少细胞外基质沉积。Background：Both mannose-6-phosphate/insulin-like growth factor Ⅱ （M6P/IGF Ⅱ ） and integrin receptors are significantly increased on the surface of activated hepatic stellate cells （HSC）. Exogenous M6P and Arg-Gly-Asp（RGD） can intervene the activation of HSC. However, few studies have been reported on the comparison or combination of the effect of these two agents. Aims： To investigate the combined interventional effects of synthetic p-aminophenyl-6- phospho-α-D-mannopyranoside （PAP-M6P） and Arg-Gly-Asp-Tyr （RGDY） on the function of activated HSC. Methods： Activated cultured HSCs at day-10 were treated with RGDY and/or PAP-M6P at different concentrations. Neither RGDY nor PAP-M6P was added in blank control group. The proliferation of HSC was measured by methyl thiazolyl tetrazolium （MTT） assay. Expression of transforming growth factor （TGF）-β1 mRNA was determined by reverse transcriptase polymerase chain reaction （RT-PCR）. Collagen type Ⅰ, hyaluronic acid and activated TGF-β1 protein were determined by enzyme- linked immunosorbent assay （ELISA）. Results： No significant difference was found in proliferation of HSC among all groups. Expressions of TGF-β1 mRNA in combination group at medium concentration and activated TGF-β1 protein in combination groups at either medium or high concentrations significantly decreased than those in RGDY control group and PAP-M6P groups at the corresponding concentrations （P〈0.05）. Collagen type Ⅰ and hyaluronic acid in combination group at high concentration were significantly decreased in comparison with that in RGDY control group and PAP-M6P groups at different concentrations （P〈0.05）. Conclusions： Combined use of PAP-M6P and RGDY can obviously reduce the expression and activation of TGF-β1 in HSC, and alleviate extracellular matrix deposition.

关 键 词：肝星状细胞 6-磷酸甘露糖 精氨酰-甘氨酰-天冬氨酸 转化生长因子Β1 胶原Ⅰ型 透明质酸 

分 类 号：R575[医药卫生—消化系统]