银杏内酯诱导嗜铬细胞瘤细胞表达低氧诱导因子-1α  

Expression of Hypoxic Inducible Factor-1α Induced by Ginkgolides in PC12 Cells

作　　者：吴小梅[1] 季秋虹[2] 顾永健[2] 朱俐[1] 

机构地区：[1]南通大学航海医学研究所,江苏南通226001 [2]南通大学附属医院神经内科,江苏南通226001

出　　处：《中国药学杂志》2006年第14期1072-1076,共5页Chinese Pharmaceutical Journal

基　　金：江苏省高校自然科学研究计划(04KJB310113);南通大学创新基金(CX0305);南通市社会发展科技计划(S5041)

摘　　要：目的研究银杏内酯(ginkgolides,Gin)对神经生长因子(nerve growth factor,NGF)诱导分化的嗜铬细胞瘤(PC12)细胞的影响以及与其相关的信号通路。方法通过MTT比色法观察不同浓度Gin对PC12细胞活性的影响,RT-PCR和Western Blot法分析Gin对PC12细胞低氧诱导因子-1(αhypoxia inducible factor-1α,HIF-1α)表达以及MAPK信号通路的影响。结果一定浓度范围的Gin可促进PC12细胞的活性,在37.5 mg.L-1作用24 h效果最明显。37.5 mg.L-1Gin单独处理PC12细胞24 h可诱导HIF-1αmRNA表达增强和蛋白水平上调。Gin还引起p-ERK水平的明显提高。Gin增加PC12细胞HIF-1α蛋白的稳定性存在一定的时间和剂量依赖关系,PD98059可部分抑制Gin的增强作用,金雀异黄素可完全阻断之;与此相对应的是,Gin以时间和剂量依赖方式诱导PC12细胞p-ERK水平的增高,PD98059和金雀异黄素均能完全阻断Gin的诱导作用。结论Gin可诱导PC12细胞表达HIF-1α,主要与MEK-ERK信号通路激活有关,可能是其促进分化的PC12生长的原因。OBJECTIVE To study the effects of ginkgolides （Gin） on the expression of hypoxia inducible factor-1α（HIF-1α） in differentiated PC12 cells induced by NGF and the involved signal pathways.METHODS The viability of PC12 cells treated with different concentration of Gin was determined by MTT assay. The expressions of HIF-1α and ERK induced by Gin were detected by Western Blot and the expression of HIF-1α mRNA was checked by RT-PCR. RESULTS Gin enhanced the viability of PC12 cells in some concentration ranges and reached the peak effect at 37.5 mg·L^-1. Both of the mRNA and the protein level of HIF- 1α were induced by the treatment with 37.5 mg·L^-1 for 24 h. Gin induced the expression of phosphory1αtion of ERK （p-ERK）. Further, Gin enhanced the stability of HIF-1α expression in a timeand dose-dependent manner. This enhancement was partially inhibited by the pretreatment with PD98059, a mitogen-activated protein kinase-1 （MEK- 1） inhibitor and completely blocked by genistein, a tyrosine kinase inhibitor. Meanwhile, the activation of p-ERK was increased in a time and dose- dependent manner by Gin and completely blocked by the pretreatment of PD98059 and genistein in PC12 cells. CONCLUSION Gin can induce the expression of HIF- 1α in PC12 cells, which may be mediated through the activation of MAPK signal pathway.

关 键 词：银杏内酯 低氧诱导因子-1Α 嗜铬细胞瘤细胞 磷酸化细胞外信号调节激酶 

分 类 号：R965[医药卫生—药理学]