EGCG对血小板源性生长因子-BB诱导的大鼠血管平滑肌细胞增殖和迁移的影响

Effects of epigallocatechin-3-gallate on the proliferation and migration of vascular smooth muscle cells induced by platelet-derived growth factor BB in rats

目的研究表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对血小板源性生长因子-BB(platelet-derived growth factor BB,PDGF-BB)诱导的血管平滑肌细胞(vascular smooth muscle cell,VSMC)增殖和迁移的影响并探讨其可能的机制。方法组织块贴壁法培养SD大鼠胸主动脉VSMC,用RTCA(real time cellular analysis)仪器检测EGCG(10、50、100μmol/L)和Jak2特异性抑制剂AG490(50μmol/L)对PDGF-BB(10 ng/mL)诱导的VSMC增殖和迁移的影响;运用Western blot检测Jak2、Stat3、Cyclin D1蛋白质水平及Stat3磷酸化水平。结果 RTCA检测结果显示,10~100μmol/L的EGCG及50μmol/L的AG490均能抑制PDGF-BB诱导的VSMC的增殖和迁移(P<0.001)。Western blot检测结果显示,50μmol/L的EGCG能明显下调PDGF-BB刺激上调的VSMC的Jak2、Stat3、Cyclin D1蛋白质水平及Stat3磷酸化(p-Stat3)水平(P<0.001),且与其他两个浓度组相比较下调更为明显(P<0.001);50μmol/L的AG490也能明显下调上述蛋白质水平(P<0.001),且50μmol/L的EGCG下调Jak2、Stat3、p-Stat3蛋白质水平与AG490组比较无明显差异。结论 EGCG抑制PDGF-BB诱导的VSMC增殖和迁移可能通过抑制Jak2/Stat3信号转导通路而实现。

Objective To investigate the effect of epigallocatechin-3-gallate （EGCG） on the proliferation and migration abilities of vascular smooth muscle cell （VSMC） induced by platelet-derived growth factor BB （PDGF-BB） and its mechanism. Methods The effects of EGCG （10, 50, 100 μmol/L） and Jak2 inhibitor AG490 on the proliferation and migration abilites of VSMC induced by PDGF-BB （10 ng/mL） in SD rats were determined by real time cellular analysis （RTCA） system. The levels of Jak2, Stst3, p-Stat3 and Cyclin D1 protiens were detected by Western blot. Results RTCA assay showed that EGCG of 10, 50, 100 μmol/L and AG490 of 50 μmol/L inhibited cell proliferation and migration of VSMC induced by PDGF-BB （P〈0.001）. Western blot showed that EGCG of 50 μmol/L down-regulated the levels of Jak2, Stat3, Cyclin D1 and p-Stat3 proteins which had been up-regulated by PDGF-BB （P〈0.001）, and the effect was better than the other two concentrations （P〈0.001）. AG490 of 50 μmol/L could also down-regulated the above mentioned proteins （P〈0.001）. There was no statistical difference in down-regulation effect between EGCG （50 μmol/L） and AG490 on the protein levels of Jak2, Stat3 and p-Stat3. Conclusions EGCG may inhibit proliferation and migration of VSMC induced by PDGF-BB through inhibiting Jak2/Stat3 signaling pathway.