RGDyK环肽对肝星状细胞增殖和迁移的影响

Effect of RGDyK peptide on the proliferation and migration of hepatic stellate cells

目的探讨RGDyK环肽对活化肝星状细胞中整合素αvβ3表达和细胞迁移的影响。方法分离SD大鼠肝星状细胞,体外传代培养诱导细胞活化,用人工合成的整合素αvβ3特异配基RGDy K环肽处理肝星状细胞,MTT法计算其生长抑制率,通过免疫化学和荧光染色,RT-PCR和Western Blot分析,与对照组比较肝星状细胞中整合素αvβ3和α-SMA的表达变化,划痕实验观察RGDy K环肽对活化肝星状细胞迁移的影响。结果与对照组相比,RGDyK环肽抑制肝星状细胞增殖并有明显的量效关系,RGDyK环肽下调肝星状细胞中整合素αvβ3和α-SMA的在mRNA和蛋白质水平的表达(P<0.05),降低肝星状细胞的迁移率(P<0.01)。结论 RGDyK环肽抑制肝星状细胞增殖、收缩和迁移等生物行为与下调活化肝星状细胞整合素αvβ3表达有关。

Objective To explore the effects of RGDyK cyclic peptide on αvβ3 integrin expression and cell migration in activated hepatic stellate cells. Methods Isolation and culture of primary hepatic stellate cell from Sprague-Dawley rat were performed and activated hepatic stellate cells model in vitro were established. RGDyK cyclic peptide was used to treat the activated hepatic stellate cells and MTT was adopted to calculate the growth inhibition rate of cells. Expression of of αvβ3 integrin expression in activated hepatic stellate cells was determined by immunochemistry staining and immunofluorescence staining , RT-PCR and Western Blot analysis of and compared between the control and treatment groups. observation the effects of cell migration was observed by wound scratche assay. Results RGDyK cyclic peptide inhibited activated hepatic stellate cells proliferation compared with the control group and a concentration-response relationship could be observed. RGDyK cyclic peptide inhibited the expression of αvβ3 integrin and a-SMA in the level of mRNA and protein in activated hepatic stellate cells respectively （P〈0.05）, and weakened the mobility of activated hepatic stellate cells （P〈0.01）. Conclusion RGDyK cyclic peptide can inhibitactivated hepatic stellate cell proliferation and migration by down-regulation of expression of αvβ3 integrin.