酪氨酸蛋白激酶抑制剂对自发性高血压大鼠血管平滑肌细胞增殖肥大的影响

Effect of tyrosine protein kinase inhibitor on proliferation and hypertrophy of vascular smooth muscle cells in SHRs

为明确自发性高血压大鼠血管平滑肌细胞(SHR VSMC)增殖与血小板源生长因子 AA (PDGF AA)、PDGF α受体表达的关系及酪氨酸蛋白激酶在其中的作用 ,我们在培养的血管平滑肌细胞中 ,采用免疫印迹 (Westernblot)、3 H TdR及3 H Leu掺入等方法 ,观察在不同来源大鼠(SHR/Wistar)血管平滑肌细胞中 ,PDGF AA、PDGF α受体及PDGF β受体表达的差异性 ;在PDGF AA刺激下细胞的增殖、肥大反应及酪氨酸蛋白激酶抑制剂 (genistein)对其的影响。结果表明 :在SHR VSMC中PDGF AA、PDGF α受体蛋白表达明显高于Wistar VSMC ,而PDGF β受体蛋白表达在SHR VSMC与Wistar VSMC无明显差异 ;在不同浓度PDGF AA(2、5、10、2 0 )ng/mL刺激下 ,SHR VSMC中PCNA表达呈剂量依赖性增高P <0 0 1;加入酪氨酸激酶抑制剂 ,SHR VSMC中PCNA表达呈剂量依赖性显著减少P <0 0 1。在不同浓度PDGF AA(2、5、10、2 0 )ng/mL刺激下 ,SHR VSMC中3 H Leu、3 H TdR掺入率呈剂量依赖性增强 ;加入酪氨酸激酶抑制剂SHR VSMC中 3 H Leu、3 H TdR掺入率显著减少。PDGF与其受体结合后 ,可激活受体细胞膜内的酪氨酸蛋白激酶 ,导致蛋白分子磷酸化的级联反应 ,启动MAPK、PKC等信号转导通路 。

Objectives To explore the relationship between proliferation, hypertrophy of vascular smooth muscle cells and PDGF-AA and PDGFR-α expression in SHRs and the role of PTK in it. Methods ①Express difference of PDGF-AA, PDGFR-α, PDGFR-β in SHR/Wistar-VSMC was observed by Western blot. ②The effect of PTK inhibitor(genistein) on proliferation and hypertrophy of SHR-VSMC induced by PDGF-AA was observed by Western blot and incorporation. Results ①PDGF-AA and PDGFR-α expression was markedly increased in SHR-VSMC than in Wistar-VSMC, but PDGFR-β was not different in SHR and Wistar-VSMC. ②Dose-dependent PDGF-AA-stimulated PCNA expression and incorporation were observed in SHR-VSMC. ③Dose-dependent genistein-inhibited PCNA expression and incorporation induced by PDGF-AA also were observed in SHR-VSMC. Conclusions Spontaneous expression increase of PDGF-AA and PDGFR-α in spontaneous hypertension rats(SHRs) may be one of the important factors on vascular reactivity and vascular modeling mediated through proliferation and hypertrophy in SHR-VSMC, and Tyrosine protein kinase plays an important role in this process.