摘要
目的通过不同剂量的奥硝唑(ORN)制备弱精子症及少弱精子症大鼠模型,并探讨其对附睾、睾丸及超微结构的损伤,以及对精子线粒体的影响。方法48只雄性SD大鼠随机分为4组,每组12只,实验组分别给予ORN 200 mg/kg、ORN 400 mg/kg、ORN 800 mg/kg灌胃,空白对照组给予1%羧甲基纤维素钠灌胃,连续灌胃28 d。观察记录各组大鼠体重、附睾及睾丸脏器指数,检测附睾精子浓度及活力,采用苏木素-伊红(HE)染色方法观察附睾头、附睾尾及睾丸病理损伤情况,透射电镜观察附睾、睾丸及精子超微结构,统计分析4组间各项指标差异。结果与空白对照组相比,ORN 200 mg/kg剂量组附睾及睾丸脏器指数、精子浓度及活力均无显著变化(P>0.05),HE染色及透射电镜结果均显示附睾及睾丸组织无明显损伤;ORN 400 mg/kg剂量组附睾脏器指数显著降低(P<0.05),睾丸脏器指数无显著变化(P>0.05),精子活力显著降低[(16.3±7.2)%vs.(36.1±6.6)%,P<0.001],精子浓度无显著变化(P>0.05),HE染色观察附睾及睾丸组织损伤不明显,透射电镜超微结构可见附睾及睾丸线粒体出现部分空泡化,精子线粒体出现损伤,但仍可见较为完整的精子线粒体结构;ORN 800 mg/kg剂量组附睾及睾丸脏器指数均显著降低(P<0.05),精子浓度[(15.3±9.2)×10^(6)/ml vs.(60.3±10.2)×10^(6)/ml,P<0.001]及活力[(7.3±5.2)%vs.(36.1±6.6)%,P<0.001]均显著下降,HE染色可见附睾及睾丸组织损伤明显,透射电镜下可观察到附睾及睾丸线粒体空泡化明显,精子线粒体明显肿胀断裂。结论ORN 400 mg/kg灌胃28 d可用于弱精子症大鼠模型的制备,其对附睾、睾丸组织损伤及精子线粒体结构损伤不明显;ORN 800 mg/kg灌胃28 d可用于少弱精子症大鼠模型的制备,可明显造成附睾及睾丸组织损伤,破坏精子线粒体结构。
Objective:To establish rat model with asthenozoospermia and oligoasthenozoospermia by administrating the different doses of ornidazole(ORN),and to explore its effects on the ultrastructural tissue damage of the epididymis and testis,and sperm mitochondria.Methods:Forty-eight male SD rats were randomly divided into 4 groups,12 rats for each group.The rats in experimental groups were given ORN 200 mg/kg,400 mg/kg and 800 mg/kg by gavage,respectively,and the negative control group was given 1%sodium carboxymethyl cellulose by gavage,continuous gavage for 28 days.The body weight,organ index of epididymis and testis,sperm concentration and motility were observed.HE staining was used to observe the pathological damage of epididymis head,epididymis tail and testis.The ultrastructure of epididymis,testis and sperm was observed by transmission electron microscope.The differences in various indicators among the four groups were analyzed.Results:Compared with the negative control group,there was no significant change in the epididymis and testis organ index,sperm concentration and motility in the ORN 200 mg/kg group,and there was no obvious tissue damage of epididymis and testis tested by HE staining and transmission electron microscopy.The epididymal organ index of ORN 400 mg/kg group significantly decreased(P<0.05),and the testicular organ index did not change significantly.Sperm motility decreased significantly[(16.3±7.2)%vs.(36.1±6.6)%,P<0.001],and sperm concentration was not changed significantly(P>0.05).HE staining showed that there was no obvious tissue damage to the epididymis and testis.Transmission electron microscopy revealed partial vacuolization of the epididymis and testicular mitochondria,and sperm mitochondria was damaged,but relatively complete sperm mitochondrial structure was still visible.In the ORN 800 mg/kg group,the epididymal and testicular organ indexes were significantly reduced,the sperm concentration[(15.3±9.2)×10^(6)/ml vs.(60.3±10.2)×10^(6)/ml,P<0.001]and motility[(7.3±5.2)%vs.(36.1±6.6)%,P<0.001]were significantly decreased.HE staining showed significant damage to the epididymis and testis.Under transmission electron microscopy,obvious vacuolization of mitochondria in the epididymis and testis can be observed,and the sperm mitochondria were significantly swollen and fractured.Conclusions:Adiministration of ORN 400 mg/kg by gavage for 28 days can be used for the establishment of rat model with asthenozoospermia,but it does not significantly damage the epididymis,testis and sperm mitochondrial structure.Adiministration of ORN 800 mg/kg by gavage for 28 days can be used for the preparation of rat model with oligoasthenozoospermia,which significantly damages the epididymis,testis and destroys the sperm mitochondrial structure.
作者
刘胜京
晏斌
安晓静
耿强
韩强
赵丰
张继伟
王福
郭军
LIU Sheng-jing;YAN Bin;AN Xiao-jing;GENG Qiang;HAN Qiang;ZHAO Feng;ZHANG Ji-wei;WANG Fu;GUO Jun(Xiyuan Hospital of China Academy of Chinese Medical Sciences,Beijing 100091;First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193;Beijing Chinese Medicine Hospital Affiliated to Capital Medical University,Beijing 100010)
出处
《生殖医学杂志》
CAS
2023年第8期1208-1215,共8页
Journal of Reproductive Medicine
基金
国家自然科学基金面上项目(82174392,82174217)
国家中医药管理局中医药传承与创新“百千万”人才工程岐黄学者资助项目(国中医药人教函〔2022〕6号)
中国中医科学院科技创新工程重大攻关项目重点项目(CI2021A02201)。
关键词
奥硝唑
男性不育症
弱精子症
少弱精子症
动物模型
Ornidazole
Male infertility
Asthenozoospermia
Oligoasthenozoospermia
Animal model
