摘要
目的探讨大麻素受体1(cannabinoid receptor 1, CBR1)在邻苯二甲酸二丁酯(dibutyl phthalate,DBP)暴露致雄性大鼠生殖损伤中的表达情况及作用机制。方法 60只健康雄性SD大鼠按随机数字表法分为4组,每组15只,分别为溶剂对照组、低剂量组(100 mg/kg)、中剂量组(250 mg/kg)和高剂量组(500 mg/kg),每日灌胃染毒1次,持续28 d建立动物模型。采用精子分析仪检测精子活力,密度,精子畸形率;HE染色法观察睾丸组织结构变化;采用ELISA检测血清睾酮(T)、雌二醇(E_2)、黄体生成素(LH)、促卵泡激素(FSH)和性激素结合蛋白(SHBG)水平;Western blot检测睾丸组织中p38、ERK、AKT和CBR1等基因的蛋白表达;RT-PCR检测相关基因mRNA的表达。结果与对照组相比,低、中、高剂量组大鼠前向运动精子(20.1±9.2),(22.3±6.2),(21.0±14.2)和精子总活力(73.6±3.2),(40.7±24.4),(41.2±4.6)显著降低,高剂量组精子密度(152.0±67.0)显著降低(P<0.05);与对照组相比,中、高剂量组大鼠血清T水平显著降低[(19.65±2.65),(17.40±3.50),P<0.05];与对照组相比,低、中、高剂量组大鼠血清中E_2(2.09±0.08),(2.11±0.77),(2.05±0.02)和LH(7.80±1.42),(7.68±2.28),(6.43±0.90)均显著降低(P<0.05),各暴露组大鼠FSH水平则显著上升[(2.88±0.37),(2.89±0.29),(3.37±0.30),P<0.05];与对照组相比,染毒组大鼠睾丸组织中CBR1的蛋白和mRNA水平均显著增加,p-p38、p-ERK水平均显著降低,差异有统计学意义(P<0.05)。结论邻苯二甲酸二丁酯暴露导致大鼠睾丸组织结构损伤和精子活力下降的机制,可能与DBP暴露致CBR1表达上调,抑制了ERK/p38MAPK的正常磷酸化有关。
Objective To explore the expression pattern of cannabinoid receptor 1(CBR1)in reproductive toxicity induced by dibutyl phthalate(DBP)exposure in male rats,and investigate the underlying mechanism.Methods Sixty healthy male Sprague-Dawley rats were randomly divided into 4 groups(15 animals in each group),that is,control group,and low-,medium-and high-dose groups,exposure to 100,250 and 500 mg/kg DBP,respectively,by intragastric injection daily for 28 consecutive days.Computer-aided sperm analysis system(CASA)was used to detect sperm vitality,concentration,and rate of deformity.HE staining was employed to observe the changes in testicular tissue.The serum levels of testosterone(T),estradiol(E2),luteinizing hormone(LH),follicle stimulating hormone(FSH)and sex hormone binding protein(SHBG)were measured by ELISA.The expression of p38,ERK,AKT and CBR1 at mRNA and protein levels was measured by RT-PCR and Western blotting.Results Compared with the control rats,the sperm motility and total sperm motility were reduced in low-(20.1±9.2 and 73.6±3.2),middle-(22.3±6.2 and 40.7±24.4)and high-dose(21.0±14.2 and 41.2±4.6)exposure groups(P<0.05).Sperm concentration was significantly lower in the high-dose group(152.0±67.0,P<0.05).Compared with the control group,serum T levels were significantly lower in the middle-(19.65±2.65)and high-dose groups(17.40±3.50,both P<0.05).The serum level of E2 was significantly decreased in each exposure group(2.09±0.08,2.11±0.77,2.05±0.02)when compared to the control group.So were the serum LH levels(7.80±1.42,7.68±2.28,6.43±0.90)(all P<0.05).While the serum level of FSH was all significantly increased in each exposure group(2.88±0.37,2.89±0.29,3.37±0.30),when compared to the control group(P<0.05).In the testis tissue,the expression of CBR1 was increased at mRNA and protein levels than that of the control group,and the expression of p-p38 and p-ERK was significantly decreased than the control group(P<0.05).Conclusion DBP exposure results in impairment of testicular tissue structure and decrease of sperm motility in male rats,which may be due to the up-regulation of CBR1 and the subsequent inhibition of ERK/p38MAPK phosphorylation.
作者
安慧慧
周妮娅
杨望
曾颖斐
陈东姣
曹佳
AN Huihui;ZHOU Niya;YANG Wang;ZENG Yingfei;CHEN Dongjiao;CAO Jia(Department of Occupational Health and Environmental Hygiene,School of Public Health,Shanxi Medical University,Taiyuan,Shanxi Province,030001;Institute of Toxicology,Faculty of Military Preventive Medicine,Army Medical University (Third Military Medical University),Chongqing,400038;Department of Occupational Health and Environmental Hygiene,College of Public Health and Management,Ningxia Medical University,Yinchuan,Ningxia Hui Autonomous Region,750004,China)
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2019年第9期836-843,共8页
Journal of Third Military Medical University
基金
国家杰出青年科学基金(81502788)~~
