环境激素联合冷刺激诱导建立慢性少精子症大鼠模型的实验研究
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摘要
目的研究环境激素联合冷刺激诱导少精子症大鼠模型的建立方法,探讨其性腺轴激素改变。方法将84只性成熟期雄性SD大鼠随机分为正常对照组(N组,12只)和造模组A(24只)、造模组B(24只)、造模组C(24只)。各造模组分别以A、B、C 3种不同环境雌激素样复合饲料联合冷刺激进行干预,24周后,通过精子浓度与活动率检测,筛选出少精子症模型,分别设为A_2、B_2、C_2组,未成模者为A_1、B_1、C_1组,观测各模型组睾丸和附睾质量与形态学改变,并行睾丸Jonhsen评分后,予以系统评价,确定最佳模型后,放免法检测其性腺轴激素水平。结果 (1)精子浓度与活动率A、B、C 3组较N组均显著降低(P<0.05,P<0.01),成模率分别为28.6%、38.1%、28.6%;(2)精子浓度、活率及运动能力A2、B2、C2组分别显著低于N组(P<0.01),B2组精子浓度显著低于A_2、C_2组(P<0.05);(3)睾丸及附睾质量A_2、B_2、C_2组均低于N组(P<0.05,P<0.01),B2组睾丸质量显著低于A2、C2组(P<0.01),组织形态发生病理改变,B2组最为明显;(4)Johnson评分A_2、B_2、C_2组均显著低于N组(P<0.01),B_2组显著低于A_2、C_2(P<0.01);(5)性激素水平检测B2组血清T水平显著低于N组(P<0.01),FSH、E2、PRL水平显著高于N组(P<0.05,P<0.01)。结论 (1)A、B、C 3种不同环境雌激素样饲料联合冷刺激均可诱导建立少精子症模型,其中B饲料联合冷刺激诱导效果最佳;(2)该方法诱导建立少精子症模型的同时,可出现精子运动力减弱,伴发弱精子症;(3)该模型的发生可能与性腺轴内分泌功能紊乱有关。
Objective To establish environmental estrogens combined with cold stimulation-induced oligospermia rat model, and analyze the changes of gonadal axis hormones. Methods Total of 84 normal male SD rats with sexual maturity were randomly divided into 4 groups, including the normal group(n=12) and the model group A, B, C(n=24). The rats in the model group A, B and C were consistently intervened respectively by different estrogen-like compound diets combined with cold stimulation. After 24 weeks, sperm oligospermia rat model was screened based on sperm concentration and sperm survival rate, they were set as group A_2, B_2, C_2, and non-model rats were set as group A_1, B_1 and C_1, and The weight and morphological changes of testis and epididymis, and Jonhson score of testis were used to determine the best model. The gonadal axis hormone level was detected by radioimmunoassay. Results The sperm concentration and sperm survival rate in group A、B、C were all lower than those in group N(P<0.05,P<0.01).The success rate of oligospermia model in intervention group was28.6%, 38.1% and 28.6%, respectively. The sperm concentration, sperm survival rate, sperm motility ability in group A_2, B_2, C_2 were all lower than those in group N(P<0.01).The sperm concentration in group B2 was lower than that in group A_2 and C_2(P<0.05).The weight of testes and epididymis in groups A2, B2, and C2 were lower than that in group N(P<0.01), and there existed a change in histological characteristics. The weight of testes in group B2 was significantly lower than that in groups A_2 and C_2(P<0.01), and the changes of pathological morphology were the most significant in group B2. The Jonhson score in group A_2, B_2, and C_2 were lower than that in group N(P<0.01).The level of T in group B_2 was lower than that in group N(P<0.01),and the levels of FSH, PRL, and E2 were higher than those in group N(P<0.05,P<0.01). Conclusion(1)A, B and C estrogen-like diets combined with cold stimulation could induce the oligospermia rat model, and B diet combined with cold stimulation had the best induction effect.(2)This method can induce oligozoospermia model and reduce sperm motility,accompanied by asthenospermia.(3)The occurrence of this model may be related to endocrine dysfunction of gonadal axis.
Objective To establish environmental estrogens combined with cold stimulation-induced oligospermia rat model, and analyze the changes of gonadal axis hormones. Methods Total of 84 normal male SD rats with sexual maturity were randomly divided into 4 groups, including the normal group(n=12) and the model group A, B, C(n=24). The rats in the model group A, B and C were consistently intervened respectively by different estrogen-like compound diets combined with cold stimulation. After 24 weeks, sperm oligospermia rat model was screened based on sperm concentration and sperm survival rate, they were set as group A_2, B_2, C_2, and non-model rats were set as group A_1, B_1 and C_1, and The weight and morphological changes of testis and epididymis, and Jonhson score of testis were used to determine the best model. The gonadal axis hormone level was detected by radioimmunoassay. Results The sperm concentration and sperm survival rate in group A、B、C were all lower than those in group N(P<0.05,P<0.01).The success rate of oligospermia model in intervention group was28.6%, 38.1% and 28.6%, respectively. The sperm concentration, sperm survival rate, sperm motility ability in group A_2, B_2, C_2 were all lower than those in group N(P<0.01).The sperm concentration in group B2 was lower than that in group A_2 and C_2(P<0.05).The weight of testes and epididymis in groups A2, B2, and C2 were lower than that in group N(P<0.01), and there existed a change in histological characteristics. The weight of testes in group B2 was significantly lower than that in groups A_2 and C_2(P<0.01), and the changes of pathological morphology were the most significant in group B2. The Jonhson score in group A_2, B_2, and C_2 were lower than that in group N(P<0.01).The level of T in group B_2 was lower than that in group N(P<0.01),and the levels of FSH, PRL, and E2 were higher than those in group N(P<0.05,P<0.01). Conclusion(1)A, B and C estrogen-like diets combined with cold stimulation could induce the oligospermia rat model, and B diet combined with cold stimulation had the best induction effect.(2)This method can induce oligozoospermia model and reduce sperm motility,accompanied by asthenospermia.(3)The occurrence of this model may be related to endocrine dysfunction of gonadal axis.
引文
1 Dubin JM,Greer AB,Kohn TP,et al.Men With Severe Oligospermia Appear to Benefit From Varicocele Repair:A Cost-effectiveness Analysis of Assisted Reproductive Technology.Urology 2018;111:99-103
2 Camargo M,Intasqui P,de Lima CB,et al.Maldi-tof fingerprinting of seminal plasma lipids in the study of human male infertility.Lipids 2014;49(9):943-956
3赵雅楠,李玉恒,李昌盛,等.不同强度冷刺激对大鼠心脏、大脑、睾丸和肺脏中CIRP表达的影响.应用与环境生物学报2012;18(01):55-59
4俞健梅,马艳萍,李永刚,等.环境内分泌干扰物对生殖健康影响的研究进展.现代生物医学进展2014;14(31):6197-6200
5陈振文,谷龙杰.精液分析标准化和精液质量评估---WHO人类精液检查与处理实验室手册(第5版)出版.中国计划生育学杂志2012;20(1):58-62
6林阳,马超,何大维,等.T3对Flu诱导隐睾大鼠生殖功能的改善研究.四川医学2017;38(3):263-266
7 SH Song,DS Kim,SH Shim,et al.Usage and perceptions of phosphodiesterase type 5 inhibitors among the male partners of infertile couples.Clini Exp Reprod Med2016;43(1):26-30
8 P Sihag,A Tandon,R Pal,et al.Sonography in male infertility:a look beyond the obvious.J Ultrasound 2018;21(3):265-276
9赵俊红.男性不育患者精液分析.医学信息2015;(31):185-185
10杜金明.126例男性无精症、少精症相关诱发因素分析.黑龙江医学2015;(3):283-284
11张卫东.氧自由基-线粒体信号通路在少、弱精子症发病中的机制研究.郑州大学2015
12揣崇,孙自学.中医药治疗少弱精子症不育的实验研究进展,广西中医、中西医结合男科学术大会、全国中西医结合男科疾病诊疗新进展学习班,2015
13陈翔,孙自学.弱少精子症证候分布规律的研究.全国中西医结合男科学术大会2017
14曾泽吟.电子垃圾拆解区环境污染对男性生殖能力的影响及相关因素分析.南方医科大学2017
15梁齐.环境雌激素影响男性生殖缺陷的研究进展.医药卫生:全文版2016;(5):00113-00113
16 Belay T,Woart A,Graffeo V.Effect of cold water-induced stress on immune response,pathology and fertility in mice during Chlamydia muridarum genital infection.Pathog Dis2017;75(5):1-13
17吕阳,王志盟,陈滨.室内空气环境与高龄者心脑血管疾病关联性研究进展.2018;34(2):124-130
18 Edenharder R,Keller G,Platt KL,et al.Isolation and characterization of structurally noveloleracea antimutagenic flavonoids from spinach.Agric Food Chem 2001;49:2767-2773
19李小梅,张丽茁,张景涛.芫荽营养与药理作用研究.黑龙江农业科学2010,22(3):121-123
20王昊,陈亮.生精障碍动物模型研究进展.中国性科学2017;26(9):102-104
21 MA Mukram,M Rafiq,SD Anturlikar,et al.Speman?AProprietary Ayurvedic Formulation,Reverses Cyclophosphamide-Induced Oligospermia In Rats.Cent Asian JGlobal Health 2013;2(1)
22马凰富,李海松,赵宗江,等.雷公藤多苷诱导生精障碍大鼠模型的建立.中华男科学杂志2015;21(2):179-184
23刘玮,康晓芳,张国巍,等.虾青素在奥硝挫致少弱精子症大鼠氧化损伤中的保护作用.中华男科学杂志2017;23(3):206-211
24熊芬,李高,庞雪冰,等.奥硝挫对雄性大鼠生育功能的影响.中华生殖与避孕杂志2006;26(2):81-85
25葛争艳,金龙,河福金,等.腺嘌呤致大鼠少精子症模型的实验研究.中国计划生育学杂志2009;17(11):659
26张才田,王雪松,胡雪玲,等.雷公藤多甙抑制大鼠精子发生的研究.生殖医学杂志2008;17(2):118-122
27 Kesari KK,Behari J.Effects of microwave at 2.45 GHz radiations on reproductive system of male rats.Toxicol Environ Chem Rev 2010;92(6):1135-1147
28蒋志惠,谢文艳,李新平,等.环境雌激素双酚A暴露现状及其雄性生殖毒性研究概况.生态毒理学报2016;11(4):1-9
29 J Peretz,JA Flaws.Bisphenol A down-regulates rate-limiting Cyp11a1 to acutely inhibit steroidogenesis in cultured mouse antral follicles.Toxicol Appl Pharmacol 2013;271(2):249-256
30 X Liu,M Miao,Z Zhou,et al.Exposure to BPA and reproductive hormones among male adults.Environ Toxicol Pharmacol 2015;39(2):934
2 Camargo M,Intasqui P,de Lima CB,et al.Maldi-tof fingerprinting of seminal plasma lipids in the study of human male infertility.Lipids 2014;49(9):943-956
3赵雅楠,李玉恒,李昌盛,等.不同强度冷刺激对大鼠心脏、大脑、睾丸和肺脏中CIRP表达的影响.应用与环境生物学报2012;18(01):55-59
4俞健梅,马艳萍,李永刚,等.环境内分泌干扰物对生殖健康影响的研究进展.现代生物医学进展2014;14(31):6197-6200
5陈振文,谷龙杰.精液分析标准化和精液质量评估---WHO人类精液检查与处理实验室手册(第5版)出版.中国计划生育学杂志2012;20(1):58-62
6林阳,马超,何大维,等.T3对Flu诱导隐睾大鼠生殖功能的改善研究.四川医学2017;38(3):263-266
7 SH Song,DS Kim,SH Shim,et al.Usage and perceptions of phosphodiesterase type 5 inhibitors among the male partners of infertile couples.Clini Exp Reprod Med2016;43(1):26-30
8 P Sihag,A Tandon,R Pal,et al.Sonography in male infertility:a look beyond the obvious.J Ultrasound 2018;21(3):265-276
9赵俊红.男性不育患者精液分析.医学信息2015;(31):185-185
10杜金明.126例男性无精症、少精症相关诱发因素分析.黑龙江医学2015;(3):283-284
11张卫东.氧自由基-线粒体信号通路在少、弱精子症发病中的机制研究.郑州大学2015
12揣崇,孙自学.中医药治疗少弱精子症不育的实验研究进展,广西中医、中西医结合男科学术大会、全国中西医结合男科疾病诊疗新进展学习班,2015
13陈翔,孙自学.弱少精子症证候分布规律的研究.全国中西医结合男科学术大会2017
14曾泽吟.电子垃圾拆解区环境污染对男性生殖能力的影响及相关因素分析.南方医科大学2017
15梁齐.环境雌激素影响男性生殖缺陷的研究进展.医药卫生:全文版2016;(5):00113-00113
16 Belay T,Woart A,Graffeo V.Effect of cold water-induced stress on immune response,pathology and fertility in mice during Chlamydia muridarum genital infection.Pathog Dis2017;75(5):1-13
17吕阳,王志盟,陈滨.室内空气环境与高龄者心脑血管疾病关联性研究进展.2018;34(2):124-130
18 Edenharder R,Keller G,Platt KL,et al.Isolation and characterization of structurally noveloleracea antimutagenic flavonoids from spinach.Agric Food Chem 2001;49:2767-2773
19李小梅,张丽茁,张景涛.芫荽营养与药理作用研究.黑龙江农业科学2010,22(3):121-123
20王昊,陈亮.生精障碍动物模型研究进展.中国性科学2017;26(9):102-104
21 MA Mukram,M Rafiq,SD Anturlikar,et al.Speman?AProprietary Ayurvedic Formulation,Reverses Cyclophosphamide-Induced Oligospermia In Rats.Cent Asian JGlobal Health 2013;2(1)
22马凰富,李海松,赵宗江,等.雷公藤多苷诱导生精障碍大鼠模型的建立.中华男科学杂志2015;21(2):179-184
23刘玮,康晓芳,张国巍,等.虾青素在奥硝挫致少弱精子症大鼠氧化损伤中的保护作用.中华男科学杂志2017;23(3):206-211
24熊芬,李高,庞雪冰,等.奥硝挫对雄性大鼠生育功能的影响.中华生殖与避孕杂志2006;26(2):81-85
25葛争艳,金龙,河福金,等.腺嘌呤致大鼠少精子症模型的实验研究.中国计划生育学杂志2009;17(11):659
26张才田,王雪松,胡雪玲,等.雷公藤多甙抑制大鼠精子发生的研究.生殖医学杂志2008;17(2):118-122
27 Kesari KK,Behari J.Effects of microwave at 2.45 GHz radiations on reproductive system of male rats.Toxicol Environ Chem Rev 2010;92(6):1135-1147
28蒋志惠,谢文艳,李新平,等.环境雌激素双酚A暴露现状及其雄性生殖毒性研究概况.生态毒理学报2016;11(4):1-9
29 J Peretz,JA Flaws.Bisphenol A down-regulates rate-limiting Cyp11a1 to acutely inhibit steroidogenesis in cultured mouse antral follicles.Toxicol Appl Pharmacol 2013;271(2):249-256
30 X Liu,M Miao,Z Zhou,et al.Exposure to BPA and reproductive hormones among male adults.Environ Toxicol Pharmacol 2015;39(2):934