补肾益气方激活哺乳动物雷帕霉素靶分子(mTOR)通路影响人早孕细胞滋养层细胞生长  

Regulatory Mechanism of the Kidney-replenishing Herb in the Proliferation of Human Cytotrophoblasts via the mTOR Pathway

作　　者：邢长英[1] 张德祥[2] 滕银成[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]江苏省南通大学附属医院,南通226001 [3]复旦大学附属妇产科医院,上海200011

出　　处：《生殖与避孕》2009年第9期584-588,597,共6页Reproduction and Contraception

基　　金：国家自然科学基金资助项目(30572446);上海市科委现代生物与制药基金资助项目(02D219115)

摘　　要：目的:研究补肾益气方对人早孕细胞滋养层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层细胞,并添加不同浓度的喂服3d补肾益气方中药的大鼠血清,培养48h,采用噻唑蓝(MTT)比色法测定细胞的增殖活力,流式细胞术检测增殖细胞核抗原(PCNA)表达、Annexin Ⅴ FITC/PI双染色法检测细胞早期凋亡。结果:与体积分数20%对照血清组(B2组)比较,体积分数10%(A1组)及20%(A2组)补肾益气方含药血清作用48h,上调细胞滋养层细胞增殖活力、促进PCNA表达、降低凋亡细胞百分率(P<0.05),上述指标均呈剂量-效应关系。哺乳动物雷帕霉素靶分子(mTOR)通路的特异性抑制剂雷帕霉素抑制20%含药血清诱导的细胞增殖活力及PCNA蛋白表达,增加凋亡细胞百分率。结论:补肾益气方对人早孕细胞滋养层细胞的生长具有促进作用,mTOR信号通路可能是中药促进细胞滋养层细胞增殖、减少细胞凋亡的主要信号转导通路之一。Objective： To investigate the mechanism of kidney-replenishing herb on the proliferation and apoptosis in human cytotrophoblasts. Methods： Human cytotrophoblasts of early pregnancy were cultured in vitro, and serum of rat which administered kidney-replenishing herb with different levels for 3 d was added for 48 h cuttured. Cell viability was monitored by MTT assay; the expression of proliferating cell nuclear antigen （PCNA） was examined by flow cytometry （FCM）, while the cell apoptosis was detected by Annexin-V stain- ing and FCM as well. Results： Compared with 20% control serum group （group B2）, the cell viability was significantly increased after treated with 10% （group A2） and 20% （group A2） herb containing serum 48 h respectively, and PCNA expression of cytotrophoblasts was up-regulated obviously, the percentages of the early apoptotic cells were significantly decreased, respectively, all of changes were in a dose-dependent manner. Rapamycin, the specific inhibitor of mTOR signal pathway inhibited the cell viability of cytotrophoblasts induced by 20% herb containing serum, not only inhibited PCNA expression of cytotrophoblasts but also the percentage of the early apoptotic cells. Conclusion： Kidney-replenishing herb has exhibited a promoting effect on the growth of human cytotrophoblasts in vitro during early pregnancy, mTOR signaling pathway may be one of the important signaling pathways in the regulating proliferation and apoptosis of cytotrophoblast ceils by the kidney-replenishing herb.

关 键 词：补肾益气方 细胞滋养层细胞 增殖 凋亡 哺乳动物雷帕霉素靶分子(mTOR) 

分 类 号：R285[医药卫生—中药学]