人工合成RGD小肽对肝星状细胞分泌细胞外基质的影响  

The Effect of Synthesized RGD Peptide on the Secretion of Extracellular Matrix (ECM) by the Hepatic Stallete Cell

作　　者：杜学亮[1] 陈颖伟[1] 王连升[1] 徐芹芳[1] 李定国[1] 吴真[1] 

机构地区：[1]上海第二医科大学附属新华医院,200092

出　　处：《放射免疫学杂志》2004年第6期433-435,共3页Journal of Radioimmanology

基　　金：国家自然科学基金资助项目 (3 0 170 412 )

摘　　要：目的 :探讨精氨酰甘氨酰天冬氨酸肽 (Arg -Gly -Asp ,RGD)对肝星状细胞分泌细胞外基质 (extra cellularmatrix ,ECM)的影响。方法 :利用化学方法合成RGD ;应用胶原酶原位灌注法分离SD大鼠原代肝星状细胞 (hepaticstallatecell,HSC) ,接种培养 4 8h后 ,随机分成 4组 :①空白对照组 ;②转化生长因子 β1(transform inggrowthfactorβ1,TGFβ1)组 :只加TGFβ1(终浓度 5ng/ml) ;③RGD组 :只加RGD(终浓度 10 0 μg/ml) ;④联合加药组 :加RGD(终浓度 10 0 μg/ml)同时加TGFβ1(终浓度 5ng/ml)。连续培养 3d后 ,放射免疫分析细胞上清液各项ECM含量变化。结果 :与空白对照组相比 ,TGF β1组和联合加药组细胞上清液中透明质酸(hyaluronicacid ,HA)、层粘连蛋白 (laminin ,LN)和Ⅲ型前胶原 (procollagenⅢ ,PCⅢ )水平均显著增高 ,RGD组细胞上清中各项ECM水平则无明显改变 ;与TGFβ1组相比 ,联合用药组HSC细胞上清LN和PCⅢ水平明显降低。结论 :RGD明显降低HSC培养上清中LN和PCⅢ含量 ,表明RGD能抑制原代HSC合成及分泌ECM ,其机制可能与竞争性结合整合素有关。Objective To study the effect of RGD peptide on the secretion of extracellular matrix by primary hepatic stallete cells. Methods The RGD peptide was synthesized by chemical method. HSCs were isolated from SD rat by in situ perfusion of liver and cultrued for 48 hours. The culture was divided into 4 parts: ① Control (no agent added) ② TGF β 1 added (TGF β 1 5ng/ml), ③ RGD added (RGD 100μg/ml), ④ Combined agents added (TGF β 1 5ng/ml and RGD 100μg/ml). Afterwards, they were further cultrued for 3 days. The levels of ECM in the culture supernatant were detected with radioimmunoassay. Results The levels of HA, LN and PCⅢ in the parts with TGF β 1 and combined agent were significantly higher than those in the control, while the levels in culture with RGD remained unchanged. The levels of LN and PCⅢ in the culture with combined agents were significantly lower than those in the culture with TGF β 1. Conclusion RGD peptide could decrease the levels of LN and PCⅢ in HSCs culture, suggesting inhibition of secretion of ECM by primary HSCs. The probable mechanism involved might be a competitive combination with integrin.

关 键 词：RGD 肝星状细胞 胞外基质 TGFβ1 PCⅢ HSC 药组 分泌细胞 连续培养 人工合成 

分 类 号：R575[医药卫生—消化系统] R318.08[医药卫生—内科学]