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目的:研究促性腺激素释放激素拮抗剂(GnRH-ant)对卵巢颗粒细胞分泌抗苗勒管激素(AMH)的影响。方法:从人卵泡液中分离、培养颗粒细胞。分别用生理盐水(NS)、顺铂(cisplatin,DDP,5μg/ml)、cetrorelix(GnRH-ant,10-9mol/L)或cetrorelix(10-9mol/L)+DDP(5μg/ml)处理颗粒细胞24 h后,ELISA方法检测颗粒细胞分泌AMH的水平。裸鼠动物随机分为4组:NS组:皮下注射NS 0.1 ml/天;1周后腹腔注射NS 0.2 ml/周。DDP组:皮下注射NS 0.1 ml/天;1周后腹腔注射5 mg/kg DDP/周。cetrorelix组:皮下注射100μg cetrorelix/天;1周后腹腔注射NS 0.2 ml/周。cetrorelix+DDP组:皮下注射100μg cetrorelix/天;1周后腹腔注射5 mg/kg DDP/周。用药第29天,经眼球取血后用ELISA方法检测各组裸鼠血清AMH的水平。结果:成功分离、培养人卵巢颗粒细胞。与DDP组及NS组比较,cetrorelix组及cetrorelix+DDP组颗粒细胞AMH分泌明显增加(P<0.05)。Cetrorelix组及cetrorelix+DDP组裸鼠血清AMH水平明显高于DDP组(P<0.05)。结论:GnRHant能够上调颗粒细胞分泌AMH,对抗DDP对AMH的降调节作用。
Objective: To study the effect of gonadotropin-releasing hormone antagonist (GnRH-ant) on the secretion of anti-Mullerian hormone (AMH) in ovarian granulosa cells. Methods: Isolated from human follicular fluid, cultured granulosa cells. The particles were treated with NS, cisplatin (DDP, 5μg / ml), cetrorelix (GnRH-ant, 10-9mol / L) or cetrorelix (10-9mol / L) After 24 hours, the level of AMH secreted by granulosa cells was detected by ELISA. Nude mice were randomly divided into 4 groups: NS group: NS 0.1 ml / day subcutaneously; NS 0.2 ml / week after 1 week. DDP group: subcutaneous NS 0.1 ml / day; 1 week after intraperitoneal injection of 5 mg / kg DDP / week. cetrorelix group: 100 μg cetrorelix injected subcutaneously per day; NS 0.2 ml / week given ip one week later. cetrorelix + DDP group: 100 μg cetrorelix per day subcutaneously; 5 mg / kg DDP per week after 1 week. On the 29th day of medication, the serum levels of AMH in nude mice of each group were detected by ELISA after blood was collected from the eyeballs. Results: Human ovarian granulosa cells were successfully isolated and cultured. Compared with DDP group and NS group, the secretion of AMH in cetrorelix group and cetrorelix + DDP group was significantly increased (P <0.05). The levels of serum AMH in Cetrorelix group and cetrorelix + DDP group were significantly higher than those in DDP group (P <0.05). CONCLUSION: GnRHant can upregulate the secretion of AMH by granulosa cells and antagonize the down-regulation of AMH by DDP.