促性腺激素释放激素拮抗剂对顺铂化疗中小鼠卵巢的保护作用

Protective effect of GnRH antagonist on ovaries of cisplatin treated mice

目的探讨促性腺激素释放激素(GnRH)拮抗剂对化疗中小鼠卵巢的保护作用。方法2018年10月至2019年10月,选取华中科技大学同济医学院动物中心购置的BALB/c雌性小鼠30只,随机数字表法分为A组、B组和C组,每组10只,其中A组腹腔注射等量生理盐水,B组腹腔注射顺铂,C组腹腔注射顺铂和西曲瑞克,采用HE染色观察各组卵巢组织,测量各组卵巢湿重,放射免疫法检测血清促卵泡生成素(FSH)、黄体生成素(LH)、雌二醇(E2)、抗苗勒管激素(AMH)和基础抑制数B(INHB)水平,蛋白质印迹法(Western blot)检测卵巢组织凋亡基因(Bax)及凋亡调控基因(Bcl-2)蛋白表达。结果B组和C组卵巢湿重明显低于A组(P<0.05),其中C组卵巢湿重为(15.17±0.88)mg,明显高于B组(8.95±0.91)mg(P<0.05);B组和C组原始卵泡、生长卵泡和成熟卵泡明显低于A组(P<0.05),其中C组原始卵泡、生长卵泡和成熟卵泡分别为(402.00±45.50)个、(110.03±21.28)个和(40.03±7.89)分,明显高于B组(230.02±40.10)个、(67.92±19.22)个和(21.11±6.92)个(P<0.05);与A组比较,B组生长卵泡及成熟卵泡明显减少,颗粒细胞排列紊乱,卵母细胞形态畸形,黄体细胞结构模糊,细胞核固缩,而C组有所改善;B组和C组FSH和LH明显高于A组(P<0.05),而E2、AMH和INHB明显低于A组(P<0.05),其中C组FSH和LH分别为(0.96±0.20)IU/L和(3.56±0.91)IU/L,明显低于B组(1.23±0.22)IU/L和(4.80±0.87)IU/L(P<0.05),而E2、AMH和INHB分别为(18.89±1.14)ng/L、(4.10±0.92)IU/L和(15.51±2.90)mg/ml,明显高于B组(9.89±1.92)ng/L、(2.20±0.54)IU/L和(12.01±2.20)mg/ml(P<0.05);B组和C组Bcl-2蛋白相对表达量明显低于A组(P<0.05),而Bax蛋白相对表达量明显高于A组(P<0.05),其中C组Bcl-2蛋白相对表达量为(0.302±0.097),明显低于B组(0.133±0.076)(P<0.05),而Bax蛋白相对表达量为(0.682±0.125),明显低于B组(0.982±0.117)(P<0.05)。结论GnRH拮抗剂可降低顺铂对小鼠卵巢的损害,保护小鼠卵巢储备功能。

Objective To investigate the protective effects of GnRH antagonist on ovarian tissue in mice after chemotherapy.Methods Technology were selected and divided into group A,B and C according to random digital table method from October 2018 to October2019,with 10 rats in each group.Group A received intraperitoneal injection of normal saline,group B received intraperitoneal injection of cisplatin,and group C received intraperitoneal injection of cisplatin and cetrorelix.HE staining was used to observe ovarian tissue in each group,the wet weight of each group was measured,serum levels of follicle stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E2),anti-mullerian hormone(AMH)and basal inhibitory number B(INHB)were measured by reflex immunoassay,western blot was used to detect the expression of Bax and Bcl-2 protein in ovarian tissue.Results The wet weight of ovaries in group B and group C were significantly lower than that in group A(P<0.05),the wet weight of ovaries in group C was(15.17±0.88)mg,significantly higher than that in group B(8.95±0.91)mg(P<0.05);The primitive follicles,growing follicles and mature follicles in group B and group C were significantly lower than those in group A(P<0.05).The scores of primitive follicles,growing follicles and mature follicles in group C were(402.00±45.50),(110.03±21.28)and(40.03±7.89),which were significantly higher than those in group B(230.02±40.10),(67.92±19.22),(21.11±6.92)(P<0.05);Compared with group A,the growth follicles and mature follicles of group B were significantly reduced,the arrangement of granulosa cells was disordered,the morphology of oocytes was abnormal,the structure of luteal cells was fuzzy,the nucleus was pyknosis,while group C was improved;The FSH and LH in group B and group C were significantly higher than those in group A(P<0.05),while E2 was significantly lower than those in group A(P<0.05).FSH and LH in group C were(0.96±0.20)IU/L and(3.56±0.91)IU/L,significantly lower than those in group B(1.23±0.22)IU/L and(4.80±0.87)IU/L(P<0.05),while E2,AMH and INHB were(18.89±1.14)ng/L,(4.10±0.92)IU/L and(15.51±2.90)mg/ml,which were significantly higher than those in group B(9.89±1.92)ng/L,(2.20±0.54)IU/L and(12.01±2.20)mg/ml(P<0.05);The relative expression of Bcl-2 protein in group B and group C were significantly lower than that in group A(P<0.05),but the relative expression of Bax protein were significantly higher than that in group A(P<0.05),the relative expression of Bcl-2 protein in group C was(0.302±0.097),significantly lower than that in group B(0.133±0.076)(P<0.05),and the relative expression of Bax protein was(0.682±0.125),was significantly lower than that in group B(0.982±0.117)(P<0.05).Conclusion GnRH antagonist can reduce the damage of cisplatin on the ovary of mice and protect the ovarian reserve function of mice.