Luo Qin, Gao Zheng-Xiang, Cao Li-Li, Yu Li, Wang Tao, Liu Han-Min
Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2010 Nov;12(11):886-90.
To investigate the role of spleen tyrosine kinase (Syk) in rat pulmonary vascular smooth muscle cells (PVSMCs) proliferation induced by platelet-derived growth factor-BB (PDGF-BB).
PVSMCs from male Sprague-Dawley rats were cultured in vitro and the cells of passages 3-5 were used in the experiment. PVSMCs were stimulated by PDGF-BB and were treated with three different doses of piceatannol, a Syk selective inhibitor. Cell proliferation was assessed by methyl thiazolyl tetrazolium (MTT) assay. DNA synthesis was measured by ³H-thymidine incorporation (³H-TdR). Cellular cycle was observed by flow cytometry. Syk mRNA and protein expression were detected using real-time quantitative PCR and Western blot, respectively.
The expression of Syk protein of PVSMCs was significantly up-regulated following PDGF-BB stimulation. PDGF-BB stimulation dramatically increased PVSMCs proliferation. After piceatannol treatment, both Syk mRNA and protein expression decreased and the proliferation of PVSMCs was inhibited in a dose-dependent manner.
Syk may promote PVSMCs proliferation induced by PDGF-BB.
探讨脾酪氨酸激酶(Syk)在血小板衍生生长因子-BB(PDGF-BB)诱导的大鼠肺血管平滑肌细胞(PVSMCs)增殖中的作用。
体外培养雄性Sprague-Dawley大鼠的PVSMCs,实验采用3-5代细胞。用PDGF-BB刺激PVSMCs,并分别用三种不同剂量的Syk选择性抑制剂碧萝芷处理。采用甲基噻唑基四氮唑(MTT)法评估细胞增殖。通过³H-胸腺嘧啶核苷掺入法(³H-TdR)测定DNA合成。采用流式细胞术观察细胞周期。分别使用实时定量PCR和蛋白质免疫印迹法检测Syk mRNA和蛋白表达。
PDGF-BB刺激后,PVSMCs的Syk蛋白表达显著上调。PDGF-BB刺激显著增加了PVSMCs的增殖。碧萝芷处理后,Syk mRNA和蛋白表达均降低,PVSMCs的增殖受到剂量依赖性抑制。
Syk可能促进PDGF-BB诱导的PVSMCs增殖。