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目的:了解血管紧张素(Ang)Ⅱ、Ang-(1-7)对人卵巢黄素化颗粒细胞的增殖及分泌功能的影响。方法:提取行体外受精/卵胞质内单精子注射-胚胎移植(IVF/ICSI-ET)患者卵泡液中的颗粒细胞,经过提纯后随机分为实验组与对照组。实验组根据添加的药物分为AngⅡ组与Ang-(1-7)组。分别向AngⅡ组/Ang-(1-7)组的颗粒细胞培养液中加入10-10~10-5 mmol/L浓度的AngⅡ/Ang-(1-7),对照组的颗粒细胞培养液中不添加药物,各组分别作用24 h、48 h、72 h,用四甲基偶氮唑蓝盐比色法(MTT法)测定颗粒细胞的吸光度(D)值;向AngⅡ组/Ang-(1-7)组的颗粒细胞培养液中分别加入10-6 mmol/L浓度的AngⅡ/Ang-(1-7),对照组的培养液中不添加药物,收集培养0~24 h、24~48 h、48~72 h的上清液,化学发光分析法(CL)测定黄素化颗粒细胞上清液中雌二醇(E2)、孕酮(P)水平。结果:不同浓度的AngⅡ、Ang-(1-7)分别作用颗粒细胞后,其D值均较对照组低,并随着浓度增加D值逐渐降低;随着时间延长,各组D值逐渐升高。10-6 mmol/L浓度的AngⅡ、Ang-(1-7)分别作用于颗粒细胞后,其上清液中E2的浓度均较对照组高,P浓度与对照组无统计学差异,随着时间延长,各组的E2和P含量逐渐增加。结论:AngⅡ及Ang-(1-7)对人卵巢黄素化颗粒细胞生长起抑制作用,其抑制作用随浓度增加逐渐增强。AngⅡ、Ang-(1-7)能够增强人卵巢黄素化颗粒细胞分泌雌激素的作用,但对孕激素的分泌没有影响。提示AngⅡ、Ang-(1-7)在人卵巢黄素化颗粒细胞的增殖及激素分泌中起一定作用,两者可能参与了人类的卵泡闭锁、优势卵泡发育、成熟以及排卵等生理过程。
Objective: To investigate the effects of Ang Ⅱ and Ang- (1-7) on the proliferation and secretion of human luteinized granulosa cells. Methods: Granulosa cells from follicular fluid of IVF / ICSI-ET patients were extracted and purified. The cells were randomly divided into experimental group and control group. The experimental group was divided into Ang Ⅱ group and Ang- (1-7) group according to the added drugs. Ang Ⅱ / Ang- (1-7) was added to Ang Ⅱ group / Ang- (1-7) group granulosa cell culture medium at a concentration of 10-10-10-5 mmol / L, and in control group granulocyte culture medium The absorbance (D) value of granulosa cells was determined by MTT method without adding any drugs for 24 hours, 48 hours and 72 hours in each group. 1-7) group were added Ang Ⅱ / Ang- (1-7) at a concentration of 10-6 mmol / L in the granulosa cell culture medium of the control group, and no drug was added to the culture medium of the control group. 48 h, 48 ~ 72 h supernatants, chemiluminescence assay (CL) determination of luteinized granulosa cell supernatant of estradiol (E2), progesterone (P) levels. Results: After treated with different concentrations of AngⅡ and Ang- (1-7), the D value of granulosa cells was lower than that of the control group, and the D value decreased with the increase of concentration. With the prolongation of time, high. The concentration of E2 in 10-6 mmol / L Ang Ⅱ and Ang- (1-7) were higher than those in control group, while the concentration of P in the supernatant was not significantly different from the control group With prolonged time, the content of E2 and P in each group gradually increased. CONCLUSION: AngⅡ and Ang- (1-7) can inhibit the growth of human luteinized granulosa cells. The inhibitory effect of AngⅡ and Ang- (1-7) increases with the increase of the concentration of Ang Ⅱ. AngⅡand Ang- (1-7) can enhance the secretion of estrogen from human luteinized granulosa cells, but have no effect on the secretion of progesterone. These results suggest that AngⅡand Ang- (1-7) play roles in the proliferation and hormone secretion of human luteinized granulosa cells, both of which may be involved in human follicular atresia, development of dominant follicles, maturation and ovulation.