摘要
目的目前黄体功能不全的机制尚不明确,文中观察补肾助孕方含药血清对大鼠垂体细胞促性腺激素(FSH)和促黄体生成激素(LH)分泌功能的影响,探讨补肾助孕方干预垂体分泌促性腺激素的作用机制。方法用补肾助孕方水煎剂给雌性SD大鼠灌胃,制备补肾助孕方含药血清。采用CCK-8法检测注射用醋酸西曲瑞克、含药血清和垂体促性腺激素释放激素(GnRH)对细胞活性的影响。在最大无毒浓度的条件下,采用注射用醋酸西曲瑞克阻滞垂体细胞GnRH受体,用RPC细胞建立受体拮抗模型(模型组),另建立空白组(正常细胞)、西曲瑞克组(仅单纯注射醋酸西曲瑞克作用6 h)、补肾助孕方组(仅加含补肾助孕药血清作用24 h)。使用20 nmol/L GnRH刺激细胞6 h,检测各组细胞上清液中FSH和LH含量,以及FSHβ与LHβ、GnRHR的mRNA表达。结果与空白组比较,西曲瑞克组细胞上清FSH降低[(3.91±0.36)m IU/m L vs(2.26±0.22)m IU/m L,P<0.05]、LH降低[(8.94±0.57)m IU/m L vs(3.35±0.59)m IU/m L,P<0.05],补肾助孕方组细胞上清LH显著升高[(8.94±0.57)m IU/m L vs(10.79±0.60)m IU/m L,P<0.05];与西曲瑞克组相比,补肾助孕方组与模型组细胞上清FSH、LH水平均明显升高(P<0.05)。与空白组比较,西曲瑞克组FSH mRNA显著降低[(0.95±0.23)vs(0.58±0.12),P<0.01]、LH mRNA显著降低[(0.98±0.14)vs(0.27±0.21),P<0.01],GnRHR mRNA表达升高[(0.97±0.13)vs(1.77±0.26),P<0.01];补肾助孕方组FSH mRNA、LH mRNA升高(P<0.05)。与西曲瑞克组比较,补肾助孕方组、模型组FSHβmRNA、LHβmRNA表达均升高(P<0.05),而GnRHR mRNA表达降低(P<0.01)。结论补肾助孕方能显著提高大鼠垂体细胞GnRH受体拮抗模型的FSH、LH分泌水平,降低异常升高的GnRH受体的mRNA及蛋白水平。补肾助孕方可改善GnRH受体表达功能。
Objective The mechanism of luteal phase defect remains unclear. To investigate the mechanism of Bu Shen Zhu Yun Decoction on the gonadotropin secretion in the pituitary gland,we observed the effects of medicated serum of Bu Shen Zhu Yun Decotion on the secretion of gonadotropin-follicle-stimulating hormone（FSH） and luteotropic hormone（LH） in rat pituitary cells.Methods The Bu Shen Zhu Yun Decotion was administered to the female SD rats by gavage to prepare the serum containing Bu Shen Zhu Yun Decoction. The CCK-8 method was used to detect the effect of cetrorelix acetate powder for injection,medicated serum and gonadotropin releasing hormone（GnRH） on cell activity. In the maximum non-toxic concentration,we used cetrorelix acetate powder for injection to block the GnRH receptor（GnRHR） in pituitary cells and established the GnRHR antagonistic model. Then we treat the model group with medicated serum（model group）. Moreover,we established the blank group（normal pituitary cells）,the cetrorelix group（intervented with cetrorelix for 6 hours）,and medicated serum group（intervented with medicated serum for 24 hours）. 20 nmol/L GnRH was used to stimulate cells for 6 h. The contents of FSH and LH in the supernatant of each group and the mRNA expression of FSHβ,LHβ and GnRHR were detected. Results Compared with that of the blank group,the supernatant levels of FSH and LH in the Cetrorelix group decreased significantly [（3. 91 ± 0. 36） m IU/m L vs（2. 26 ± 0. 22） m IU/m L,（8. 94 ± 0. 57） m IU/m L vs（3. 35 ±0.59） m IU/m L,P〈0.05） ]. In contrast,the levels of LH significantly increased [（8.94±0.57） m IU/m L vs（10.79±0.60） m IU/m L,P〈0.05） ]; Compared with the cetrorelix group,the levels of FSH and LH in both medicated serum group and model group increased significantly（P〈0.05）. Compared with the blank group,the mRNA level of FSH and LH in the cetrorelix group decreased significantly[（0.95±0.23） m IU/m L vs（0.58±0.12） m IU/m L,（0.98±0.14）
作者
刘贝
周惠芳
周伯如
蒋小飞
徐建亚
谢彤
单进军
LIU Bei;ZHOU Hui-fang;ZHOU Bo-ru;JIANG Xiao-fei;XU Jian-ya;XIE Tong;SHAN Jin-jun(The First Clinical Medical College,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China;Institute of Pediatrics,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China)
出处
《医学研究生学报》
CAS
北大核心
2018年第7期703-708,共6页
Journal of Medical Postgraduates
基金
国家自然基金(81473713)
中国和欧洲的医疗保健业发展合作研究计划(FP7/2007-2013/PIRSES-GA-2013-612589)
江苏省自然基金(BK20141463)
