脾酪氨酸激酶对大鼠肺血管平滑肌细胞增殖的作用

Effect of spleen tyrosine kinase on the proliferation of pulmonary vascular smooth muscle cells in rats

摘要:

目的：研究脾酪氨酸激酶（spleen tyrosine kinase, Syk）在血小板源性生长因子BB（platelet-derived growth factor-BB, PDGF-BB）诱导大鼠肺血管平滑肌细胞（PVSMCs）增殖中的作用。方法：体外培养雄性Sprague-Dawley大鼠PVSMCs，经鉴定后选用3~5代细胞，经PDGF-BB诱导细胞增殖后，用3种不同剂量Syk选择性抑制剂piceatannol干预，以MTT比色法测定细胞增殖，3H-TdR掺入法检测细胞DNA合成，流式细胞仪检测细胞周期，荧光定量PCR、Western blot分别检测细胞Syk mRNA和蛋白表达水平。结果：PDGF-BB刺激后，Syk蛋白表达水平明显增高，细胞显著增殖；经Syk抑制剂piceatannol干预后，随着Syk mRNA和蛋白表达水平的降低，细胞的增殖受到明显抑制，其抑制作用与剂量大小具有相关性。结论：在PDGF-BB诱导下的大鼠肺血管平滑肌细胞增殖中，Syk可能起促进作用。 ［中国当代儿科杂志，2010，12（11）：886-890］

Abstract:

OBJECTIVE: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: Syk may promote PVSMCs proliferation induced by PDGF-BB.［Chin J Contemp Pediatr, 2010, 12 (11):886-890］