D-半乳糖诱导迟发性性腺功能减退大鼠模型的建立与评价  

Establishment and evaluation of a rat model of late-on-set hypogonadism using D-galactose

作　　者：宁港 吴悔 李波男 石若冰 王浩宇 周兴[1] NING Gang;WU Hui;LI Bo-nan;SHI Ruo-bing;WANG Hao-yu;ZHOU Xing(Department of Andrology,The First Afilited Hospital of Hunan Universilty of Chinese Medicine,Changsha,Hunan 410007,China;Graduate School,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Hunan University of Traditional Chinese Medicine,Changsha,Hnnan 410208,China)

机构地区：[1]湖南中医药大学第一附属医院男科,湖南长沙410007 [2]湖南中医药大学研究生院,湖南长沙410208 [3]湖南中医药大学,湖南长沙410208

出　　处：《中华男科学杂志》2022年第8期675-684,共10页National Journal of Andrology

基　　金：国家自然科学基金(82074444,81673984);湖南创新型省份建设专项经费资助-“湖湘青年英才”支持计划(2019RS2041);湖南省教育厅重点项目(20A368);长沙市科技计划项目经费资助-长沙市杰出创新青年培养计划(kq1802015);湖南省高层次卫生人才225工程医学学科骨干人才培养对象(湘卫函[2019]196号);中华中医药学会“青年人才托举工程”(2019-QNRC2-C07);湖南中医药大学中西医结合一流学科开放基金(2020ZXYJH64)~~。

摘　　要：目的:探讨D-半乳糖(D-gal)复制迟发性性腺功能减退(LOH)大鼠模型的可行性及其最佳造模浓度与造模时长,为LOH基础研究提供可参考模型。方法:35只8周龄雄性SD大鼠随机分为正常对照组,6周D-gal低、中、高剂量组,8周D-gal低、中、高剂量组。D-gal高、中、低剂量组分别腹腔注射480、240、120 mg/(kg·d)D-gal,6周组连续注射42 d,8周组连续注射56 d,正常对照组大鼠每日腹腔注射相同体积生理盐水,18月龄SD大鼠5只视为阳性对照组。实验结束后,进行大鼠悬尾试验和交配实验。称重后处死,采血检测空腹血糖,离心后取上清液酶联免疫法检测睾酮(T)水平,取双侧睾丸称重计算睾丸指数,取附睾收集精液用精子质量检测系统检查精液常规,取睾丸组织进行Western印迹检测BMAL1、NR1D1蛋白表达,以光学显微镜和透射电子显微镜观察睾丸组织的病理变化与超微结构。结果:与正常对照组比较,造模后大鼠血清T水平,6周D-gal中剂量组、6周D-gal高剂量组、8周D-gal中剂量组、8周D-gal高剂量组、阳性对照组均明显降低(P<0.05);精子浓度、前向运动精子百分率,8周D-gal高剂量组、阳性对照组均显著降低(P<0.05);8周D-gal低、中、高剂量组体重均显著增加(P<0.05);扑捉次数、射精次数各剂量造模组、阳性对照组均明显降低(P<0.05);悬尾不动时间6周D-gal高剂量组和8周D-gal低、中、高剂量组及阳性对照组均显著升高(P<0.05);空腹血糖各剂量造模组、阳性对照组均明显升高(P<0.05);睾丸组织中BMAL1蛋白表达量,6周D-gal高剂量组和8周D-gal低、中、高剂量组及阳性对照组均明显降低(P<0.05);NR1D1蛋白表达量,8周D-gal中、高剂量组及阳性对照组均明显降低(P<0.05)。与阳性对照组比较,大鼠血清T水平,6周D-gal高剂量组、8周D-gal高剂量组无显著差异(P>0.05);睾丸指数8周D-gal中、高剂量组无显著差异(P>0.05);精子浓度、前向运动精子百分Objective: To explore the feasibility and duration of establishing a model of late-on-set hypogonadism(LOH) in rats using D-galactose(D-gal) and the optimal concentration of D-gal in modeling. Methods: Thirty-five 8-week-old male SD rats were randomly divided into seven groups of an equal number to receive intraperitoneal injection of normal saline(normal control) or D-gal at 480 mg/kg/d(high-dose), 240 mg/kg/d(medium-dose) and 120 mg/kg/d(low-dose) for 6 or 8 weeks respectively. Another five 18-month-old SD rats were taken as positive controls. After modeling, the animals were subjected to tail suspension test and mating test, weighed, and then sacrificed for examination of the levels of fasting blood glucose(FBG) and serum testosterone(T), testis index and semen parameters, determination of the expressions of the BMAL1 and NR1 D1 proteins in the testis tissue by Western blot, and observation of the pathological and ultrastructural changes in the testis tissue under the light microscope and transmission electron microscope. Results: Compared with the normal controls, the serum T level was significantly decreased in the 6-week medium-dose D-gal(6 WMD), 6-week high-dose D-gal(6 WHD), 8 WMD, 8 WHD and positive control groups(P < 0.05), and so were sperm concentration and the percentage of progressively motile sperm(PMS) in the 8 WHD and positive control groups(P < 0.05), and the numbers of captures and ejaculations in all the model rats and positive controls(P < 0.05), while the body weight was markedly increased in the 8-week low-dose D-gal(8 WLD), 8 WMD and 8 WHD groups(P < 0.05), and so were the tail suspension time in the 6 WHD, 8 WHD, 8 WMD, 8 WLD and positive control groups(P < 0.05) and the FBG level in all the model rats and positive controls(P < 0.05). The expression of the BMAL1 protein was significantly decreased in the 6 WHD, 8 WLD, 8 WMD, 8 WHD and positive control groups(P < 0.05), and so was that of NR1 D1 in the 8 WMD, 8 WHD and positive control groups(P < 0.05). Compared with the positive controls,

关 键 词：迟发性性腺功能减退 大鼠模型 D-半乳糖 

分 类 号：R698[医药卫生—泌尿科学]