补肾益气方激活ERK通路影响人早孕细胞滋养细胞生长  

The ERK pathway regulates human cytotrophoblasts proliferation induced by kidney-replenishing herb

作　　者：邢长英[1] 张德祥[2] 滕银成[1] 陆丽华[1] 归绥琪[3] 

机构地区：[1]上海交通大学附属第六人民医院,上海200233 [2]南通大学附属医院 [3]复旦大学附属妇产科医院

出　　处：《现代妇产科进展》2009年第4期273-277,共5页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金资助项目(No:30572446);上海市科委现代生物与制药基金资助项目(No:02D219115)

摘　　要：目的:研究补肾益气方对人早孕细胞滋养细胞增殖和早期凋亡的影响及其可能的信号转导通路。方法:体外分离培养人早孕细胞滋养细胞,用四甲基偶氮唑盐(MTT)比色法测定细胞的增殖活力,流式细胞术增殖细胞核抗原(PCNA)表达检测细胞增殖、AnnexinⅤFITCPI双染色法检测细胞早期凋亡,Western blot法检测细胞外信号调节蛋白激酶1/2(ERK1/2)磷酸化。结果:与20%对照血清组比较,10%及20%补肾益气方含药血清作用48h,上调细胞滋养细胞增殖活力,促进PCNA表达、降低早期凋亡细胞百分率、诱导细胞ERK磷酸化,上述指标均呈量-效关系;ERK通路的特异性抑制剂U0126几乎完全抑制20%含药血清诱导的细胞ERK1/2活化,下调20%含药血清诱导的细胞增殖活力及PCNA蛋白表达、增加早期凋亡细胞百分率。结论:补肾益气方对人早孕细胞滋养细胞的生长具有促进作用,ERK1/2信号通路可能是补肾益气方促进细胞滋养细胞增殖、减少细胞凋亡的主要信号转导通路之一。Objective： To investigate the intracellular signaling pathway involved in the mediation of human first-trimester cytotrophoblast proliferation and apoptosis induced by kidneyreplenishing herb. Methods：The first-trimester human cytotrophoblast was cultured in vitro for 48h, with either control serum or serum made from kidney-replenishing herb. The cell viability was monitored by MTT assay. The cell proliferation, proliferating cell nuclear antigen （PCNA） was detected by flow cytometry（FCM）. The cell apoptosis was detected by Annexin-V ,FITC PI double staining and FCM. The level of phosphorylated form of extracellular signal-regulated protein kinase1/2（ERK1/2） was detected by Western blot. Results： Compared with 20% control serum group,the cell viability and PCNA expression of cytotrophoblast were significantly increased after treated with 10% and 20% herb serum for 48h respectively. The percentage of the early apoptotic cell was significantly decreased, and the ratio of phosphorylated ERK （pERK） to total ERK expression was increased induced by the herb serum. All of the regulations of these markers were in a dose-dependent manner. The specific inhibition of ERK1/2 using U0126 inhibited phosphorylated ERK1/2 induced by 20% herb serum. U0126 not only inhibited the cell viability and PCNA expression of cytotrophoblast, but also enhanced the apoptosis of cell. Conclusion： Kidney-replenishing herb improves the growth of human first-trimester cytotrophoblast in vitro maybe through activation of ERK1/2 pathway.

关 键 词：补肾益气方 植物药疗法 细胞滋养细胞 增殖 细胞凋亡 细胞外信号调节蛋白激酶 

分 类 号：R714.14[医药卫生—妇产科学]