RGDS四肽对肝星状细胞增殖、胶原合成及FAKmRNA表达的影响

Inhibitory effects of Arg-Gly-Asp-Ser(RGDS) tetrapeptide on proliferation and collagen synthesis via downregulating focal adhesion kinase in hepatic stellate cell

目的研究精-甘-天冬-丝氨酸(RGDS)四肽对大鼠肝星状细胞(HSC)增殖和胶原合成的抑制作用以及对黏着斑激酶(FAK)的影响。方法以不同剂量的RGDS干预纤维黏连蛋白(FN)刺激的HSC,通过甲苯胺蓝染色测定细胞黏附率,3H-胸腺嘧啶(3H-TdR)、3H-脯氨酸(3H-Pro)掺入法测定HSC增殖及胶原合成能力,RT-PCR检测FAKmRNA的变化。结果与单纯FN组相比较,不同剂量(25mg/L、50mg/L和100mg/L)的RGDS作用于HSC48h及100mg/L的RGDS作用于HSC不同时间(12h、24h和48h)后,HSC的增殖明显受抑,胶原合成能力降低,同时FAKmRNA表达水平也下降。结论RGDS能够抑制FN与HSC黏附,并抑制HSC增殖及胶原合成。FAK的下调可能是RGDS抑制HSC增殖及胶原合成的分子机制之一。

Objective To investigate the inhibitory effect of Arg-Gly-Asp-Ser （RGDS） on proliferation and collagen synthesis stimulated by fibronectin （FN） and the influence on the expression of focal adhesion kinase of hepatic stellate cell （HSC）, in order to provide a theoretical ground for its use in prevention and treatment of hepatic fibrosis in clinic. Methods The adhesion rates were determined by toluidine blue colorimetric assay. The effects of RGDS on the proliferation and collagen synthesis of HSC were assessed by ^3H-TdR and ^3H-proline incorporation assays, respectively. The FAK mRNA of HSCs was determined by RT-PCR Results ①RGDS was found to inhibit the rates of adhesion, while RGES has no such ability; ②The proliferation and collagen synthesis of HSC were inhibited by RGDS in dose and time-dependent manners; ③The expression of FAK in the （25, 50, 100mg/L） RGDS groups was significantly lower than that in the control group at mRNA level. Conclusions RGDS can inhibit proliferation and collagen synthesis of HSC, and these effects correlate with its anti-adhesion ability. Meanwhile the expression of FAK is also down-regulated significantly. As a signaling molecule, FAK may be involved in the effect of RGDS. This may be one of the underlying mechanisms of anti-fibrosis effect of RGDS.