论文部分内容阅读
Vascular endothelial growth factor (VEGF)-A/VEGFR-1 (Flt-1)/VEGFR-2 (KDP/Flk-1) signal system regulate physiological as well as pathological angiogenesis.Our previous study showed that Y-27632 or HA-1077,Rho-associated kinase (ROCK) specific inhibitors, inhibited VEGF-A and Kdr protein expression.In this study, it was investigated that the relationship between ROCK and VEGF-A and related receptors using a gene silencing.BMSCs derived from Sprague-Dawley rats were induced to differentiate into endothelial like cells (rBMSC-ECs)by HG-DMEM plus EGM-2.Gene silence of ROCK-1 and ROCK-2 was performed by small interference RNA (siRNA) of ROCK-1 or ROCK-2 transfected into the rBMSC-ECs with lipofectamine 2000 respectively.The mRNA expression levels were analyzed by quantitative real time PCR (qRT-PCR).The results showed that the mRNA level of ROCK-1 and ROCK-2 were decreased by 70±0.7% (P<0.01) and 80±0.6% (P<0.01) compared with the group of control respectively after 48h.Meanwhile, in the group of rBMSC-ECs after transfected siRNA ROCK-1, the mRNA level of VEGF-A was increased 80±0.2% (P<0.05), Fit-1 and Kdr decreased 40±0.1% (P<0.05)and 70±0.3% (P<0.05) respectively compared with the control group.However, in rBMSC-EC after transfected siRNA ROCK-2, the mRNA level of VEGF-A and Flt-1 were decreased 30%±0.2 (P<0.05) and 50±0.1% (P<0.05)respectively, but Kdr increased 20±0.2% compared with control group.These results suggested that the role of ROCK-1 and ROCK-2 was different in VEGF/VEGFR signal system.The gene expression of VEGF-A was upregulated, and Kdr and Fit-1 were down-regulated when the gene of ROCK-1 was silenced.The gene expression of VEGF-A and Flt-1 were down-regulated, and Kdr was up-regulated slightly when the gene of ROCK-2 was silenced.