抑制miR-29a表达对体外蜕膜化人子宫内膜基质细胞蜕膜催乳素分泌的影响  

Effect of inhibiting miR-29a expression on prolactin secretion of human endometrial stromal cells during in vitro decidualization

作　　者：张鹏[1] 姜万君 

机构地区：[1]中南大学生殖与干细胞工程研究所,长沙410078 [2]南昌大学医学院

出　　处：《山东医药》2016年第7期1-3,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(31260248)

摘　　要：目的观察抑制miR-29a表达对体外蜕膜化人子宫内膜基质细胞蜕膜催乳素(d PRL)分泌水平的影响。方法收集子宫良性病变患者全子宫切除术中留取的子宫内膜组织,分离子宫内膜基质细胞。取部分细胞分为实验组与对照组,实验组用雌二醇(E2)和孕酮(P4)对细胞进行蜕膜化处理,对照组不进行蜕膜化处理;取部分细胞按2×105/m L接种到6孔板中,分为1、2、3组,1组转染anti-miR-29a,2组转染control miRNA,3组为空白对照;分别于培养0、24、48、72 h后收集各组细胞。采用real-time PCR法检测各组细胞中的miR-29a,采用化学发光法检测各组细胞培养液上清中的d PRL。结果实验组培养0、24、48、72 h细胞中miR-29a相对表达量及细胞培养液上清中d PRL水平高于对照组(P均<0.01);实验组随培养时间延长,miR-29a表达逐渐上调,培养液上清中d PRL水平增高,各培养时点两两相比,P均<0.05。培养24、48、72 h时1组细胞中miR-29a相对表达量低于2、3组,细胞培养液上清中d PRL水平低于2、3组(P均<0.01)。结论抑制miR-29a表达后,蜕膜化人子宫内膜基质细胞分泌d PRL减少;miR-29a可能参与了子宫内膜基质细胞的蜕膜化过程,并调控子宫内膜基质细胞分泌d PRL。Objective To observe the effect of inhibiting miR-29 a expression on prolactin secretion of human endometrial stromal cells（ HESCs） during in vitro decidualization. Methods We collected the endometrial tissues during panhysterectomy from patients with uterine benign lesions,and used trypsin method to isolate HESCs. The cells were then divided into the experimental group and control group,the experimental group was treated with estradiol（ E2） and progesterone（ P4）,and the control group was not treated. We took part of the cells in 2 × 10^5/ m L to inoculate into the 6-well plates and and then divided them into groups1,2 and 3. The group 1 was transfected by anti-miR-29 a. Group 2 was transfected by Control-miRNA and group 3 as taken as the blank control. After being cultured for 0 h,24 h,48 h,72 h we collected cells in each group. real-time PCR was used to detect miR-29 a in the cells of each group. d PRL in the supernatant of cell culture medium was detected by chemiluminescence method. Results The relative expression of miR-29 a in the cells after being cultured for 0 h,24 h,48 h,72 h,and the d PRL level in the cell culture supernatantthe of the experimental group were higher than those of the control group（ all P〈0. 01）. In the experimental group,with the prolonged culture time,the miR-29 a expression was gradually increased,and the level of d PRL in the culture supernatant was increased. Significant difference was found between every two time points（ all P〈0. 05）. The relative expression of miR-29 a in the group 1 at 24 h,48 h and 72 h was lower than that in the groups 2 and 3,and the level of d PRL in the supernatant of group 1 was lower than that in the groups 2 and 3（ all P〈0. 01）. Conclusions After the inhibition of miR-29 a expression,human decidual endometrial stromal cells reduce the secretion of d PRL. miR-29 a may be involved in the decidual process of HESCs,andregulates the endometrial stromal cells secreting dP RL.

关 键 词：微小RNA-29a 蜕膜催乳素 人子宫内膜基质细胞 

分 类 号：R711.51[医药卫生—妇产科学] R321.3[医药卫生—临床医学]