骨形态发生蛋白-9对黄素化颗粒细胞增殖的影响  

Effects of bone morphogenetic protein 9 on the proliferation of luteinized granulosa cells cultured in vitro

作　　者：刘灿 汪辉 甄秀丽[1] 任泽亚 张坪[1] 江乾[1] 郝雅丽[1] LIU Can;WANG Hui;ZHEN Xiuli(Reproductive Medical Center,The Forth Hospital of Hebei Medical University,Hebei,Shijiazhuang 050011,China)

机构地区：[1]河北医科大学第四医院生殖医学科,石家庄市050011

出　　处：《河北医药》2018年第17期2573-2577,共5页Hebei Medical Journal

基　　金：河北省科学技术研究与发展支撑计划项目(编号:16277774D)

摘　　要：目的检测外源性添加骨形态发生蛋白-9(BMP-9)基因重组蛋白对体外培养黄素化颗粒细胞增殖的影响。方法因输卵管因素行体外受精-胚胎移植患者90例的卵泡液,分离提取黄素化颗粒细胞,体外培养添加重组BMP-9的试验组,采用CCK-8,实时定量PCR检测颗粒细胞的增殖情况。结果体外培养24、48、72 h,对照组与试验组颗粒细胞均呈稳定增殖,添加BMP9重组蛋白可显著促进人卵巢黄素化颗粒细胞的体外增殖,CCK-8试剂盒测定吸光度值试验组均高于对照组,差异有统计学意义(P<0.05),其中体外培养48 h与对照组比较颗粒细胞增殖率最高(P<0.05)。BMP-9基因重组蛋白能够显著上调PCNAmRNA的表达水平(P<0.05)。结论外源性添加BMP-9能够促进体外培养人卵巢黄素化颗粒细胞的增殖。Objective To investigate the effects of exogenous bone morphogenetic protein 9（ BMP 9）,genetic recombination protein on the proliferation of luteinized granulosa cells cultured in vitro. Methods A total of 90 cases of follicular fluid from patients who underwent in vitro fertilization（ IVF） embryo transfer because of fallopian tube obstruction in our hospital from July 2017 to December 2017 were enrolled in the research. The luteinized granulosa cells of patients were separated and extracted,which were cultured in medium added with recombinant BMP 9 in vitro and to be regarded as experimental group,however,those without treatment with BMP 9 were regarded as control group. The proliferation status of granulosa cells was measured by means of CCK 8 and real time quantitative PCR,respectively. Results There existed a stable proliferation status of luteinized granulosa cells in both group after cultured in vitro for 24 h,48 h,72 h. The recombinant BMP 9 protein could significantly promote the proliferation of the human ovarian luteinized granulose cells. The optical density（ OD） value that was detected by using CCK 8 kit in experimental group was significantly higher than that in control group（ P 〈 0. 05）. The proliferation rate of granulose cell at 48 h in experimental group was significantly higher than that in control group（ P 〈 0. 05）. Moreover the genetic recombinant BMP 9 protein could significantly upregulate the expression levels of proliferating cell nuclear antigen（ PCAN） mRNA（ P 〈 0. 05）. Conclusion The exogenous addition of BMP 9 can promote the proliferation of human ovarian luteinized granulosa cells cultured in vitro.

关 键 词：骨形态发生蛋白-9 体外受精-胚胎移植 颗粒细胞 

分 类 号：R321.2[医药卫生—人体解剖和组织胚胎学]