3-甲基-4-硝基酚对大鼠睾丸组织的损伤作用

为从病理学角度探讨3-甲基-4-硝基酚(PNMC)的生殖毒性,采用24只SPF雄性SD大鼠随机分为4组,染毒组分为100 mg/kg体重、10 mg/kg体重和1 mg/kg体重三组,连续5 d皮下注射,对照组皮下注射等量的溶剂(PBS+0.05%tween80)。最后一次染毒后24 h,无痛处死大鼠,称取睾丸湿重并计算其指数。将已固定的睾丸组织进行石蜡切片,采用H.E和免疫组化染色的方法观察PNMC对大鼠睾丸的病理损伤及其对凋亡相关蛋白Bcl-2和Bax表达的影响。结果显示各个染毒组与对照组相比较,睾丸大体湿重和睾丸指数明显下降(P0.05或P0.01);H.E染色结果显示各试验组睾丸曲细精管生精细胞均有不同程度的空泡变性兼或坏死;免疫组织化学染色观察发现:各实验组睾丸中Bax蛋白表达量显著高于对照组(P0.01),而Bc1-2蛋白表达量却明显低于对照组(P0.01),Bc1-2/Bax也显著低于对照组(P0.01)。研究结果表明,PNMC不仅可引起雄性大鼠睾丸曲细精管生精细胞变性、坏死,还可促进生殖细胞的大量凋亡。     

3-甲基-4-硝基酚对大鼠肾脏氧化损伤作用的研究

为了探讨3-甲基-4-硝基酚(PNMC)对大鼠肾脏的氧化损伤作用,选择SPF级雄性SD大鼠,随机分为4组(每组10只),设立3个染毒组和1个对照组。染毒组采用皮下注射的途径分别给予每天剂量为1 mg/kg体重、10mg/kg体重和100 mg/kg体重的PNMC,连续5 d。对照组采用同样的途径给予溶剂(PBS+0.05%tween80),最后一次染毒后第24小时剖检取材。称量体重和器官重量,观察肾脏组织病理学变化,检测血清中肌酐和尿素氮的含量,以及超氧化物歧化酶、谷胱甘肽过氧化物酶、总抗氧化能力及丙二醛等抗氧化指标的含量。结果显示,染毒组大鼠体重和日增重出现不同程度的下降;各剂量组大鼠肾脏均有不同程度的病理损伤;100 mg/kg体重组大鼠肌酐浓度显著升高,所有染毒组大鼠超氧化物歧化酶的含量均显著下降,10 mg/mg体重染毒组大鼠总抗氧化能力也显著低于对照组。综上所述,PNMC染毒后血清中超氧化物歧化酶的活性降低,同时伴有总抗氧化能力下降,提示大鼠体内抗氧化系统遭到破坏,导致过多的过氧化物在体内蓄积,从而对大鼠肾脏造成氧化损伤。     

不同运动方式和运动时间对小鼠睾酮分泌和精子生成的影响

为深入探究不同运动方式和运动强度对正常雄性动物生殖机能的影响,本研究以C57BL/6J小鼠为研究模型,将32只11周龄雄性小鼠随机分为对照组、(EE)组、短期有氧运动(AAE)和长期有氧运动(CAE)组,运动结束后采集血液、睾丸、附睾样品,检测血清睾酮水平,分析睾酮曲细精管管腔面积和生精标志基因表达、精子形态等差异。结果表明:与对照组小鼠相比,EE组小鼠血清睾酮水平显著降低,睾丸类固醇转运蛋白STAR的mRNA表达显著降低,且睾丸曲细精管和附睾管管壁形态不完整,精子总数降低,精子畸形率显著增加;AAE组小鼠睾丸发育及精子品质与对照组接近,但CAE组小鼠血清睾酮水平较对照组显著升高,睾丸类固醇转运蛋白STAR的mRNA表达水平显著增加,睾丸曲细精管空腔面积显著降低,精子畸形率显著降低。上述结果说明,长期有氧运动是提高小鼠生殖机能的有效方式,短期有氧运动无此效应,急性力竭运动有损小鼠的生殖能力。     

柠檬酸对小鼠精子特性和睾丸组织形态结构的影响

60只雄性小鼠随机分为4组,每天分别腹腔注射0.2mL的0.9%生理盐水或7.5、15、30g/L CA,于14、21d测定曲细精管精子的生成及附睾内精子特性的相关数据。结果显示,随柠檬酸处理剂量的增加精子的生成数减少。14d时,对照组与15、30g/L CA组间、7.5、30g/L CA组间曲细精管内精子的生成差异显著（P〈0.05）;21d时,对照组与柠檬酸处理组间及7.5、15g/L CA与30g/L CA处理组间曲细精管内精子的生成有显著差异（P〈0.05）。柠檬酸对附睾精子品质的影响表现为：对照组与柠檬酸处理组间及柠檬酸处理组间精子密度和活动率差异均显著（P〈0.05）;对照组和15、30g/L CA处理组及15g/L CA与30g/L CA处理组间精子活力差异显著（P〈0.05）;精子的畸形率随柠檬酸浓度的增加而增加,15g/L CA处理组精子畸形率显著高于对照组（P〈0.05）,30g/LCA处理组显著高于对照组和7.5g/L CA（P〈0.05）处理组。组织形态学观察结果表明柠檬酸能破坏睾丸正常组织形态结构。柠檬酸高剂量组的生精小管内生精细胞排列疏松、紊乱,生精细胞间出现空隙;生精小管中的生精细胞脱落或退化,精原细胞、精母细胞和精子数量明显减少。研究表明,外源柠檬酸雄性小鼠具有显著的生殖遗传毒性。     

亚急性硒中毒对小鼠精子质量的影响研究

为了研究亚急性硒中毒对小鼠精子质量的影响,试验选用雄性昆明鼠灌胃染毒。高、中、低剂量组分别按照0.135 mg/kg、0.540 mg/kg、2.160 mg/kg剂量浓度灌胃亚硒酸钠溶液,对照组灌胃纯化水,连续攻毒30 d,每隔10 d各组随机选取10只采样,检查睾丸脏器系数、精子密度、精子活率、精子畸形率。随着染硒剂量的增加和时间的延长,各染硒剂量组睾丸脏器系数呈下降趋势,小鼠精子密度和精子活率随染硒剂量的增加逐渐减小,各染硒组小鼠的精子畸形率均有增加,呈现明显的时间-剂量-效应关系。结果表明：亚急性硒中毒可影响雄性小鼠生长发育,阻碍睾丸的发育甚至引起睾丸萎缩,影响精子的生成和发育,降低雄性小鼠的生殖机能。     

亚急性硒中毒对小鼠精子质量的影响研究

为了研究亚急性硒中毒对小鼠精子质量的影响,试验选用雄性昆明鼠灌胃染毒。高、中、低剂量组分别按照0.135 mg/kg、0.540 mg/kg、2.160 mg/kg剂量浓度灌胃亚硒酸钠溶液,对照组灌胃纯化水,连续攻毒30 d,每隔10 d各组随机选取10只采样,检查睾丸脏器系数、精子密度、精子活率、精子畸形率。随着染硒剂量的增加和时间的延长,各染硒剂量组睾丸脏器系数呈下降趋势,小鼠精子密度和精子活率随染硒剂量的增加逐渐减小,各染硒组小鼠的精子畸形率均有增加,呈现明显的时间-剂量-效应关系。结果表明:亚急性硒中毒可影响雄性小鼠生长发育,阻碍睾丸的发育甚至引起睾丸萎缩,影响精子的生成和发育,降低雄性小鼠的生殖机能。     

氟致小鼠睾丸组织形态结构损伤的观察

选取20只20日龄的昆明小鼠,随机分为4组,分别给予含0,50,100和150 mg/L氟化钠的饮水120 d,通过石蜡组织切片观察氟对睾丸的损伤作用。结果发现,小鼠饮用含有氟化钠的水后,曲细精管管壁断裂、结构排列紊乱;生精细胞、次级精母细胞从生精上皮脱落;精母细胞胞质和胞核中出现大量的空泡样病变,且细胞的微核率增加;精子出现畸形。这说明氟对睾丸组织结构具有明显的损伤作用。     

氟暴露小鼠睾丸形态计量学研究

为了观察小鼠氟中毒睾丸形态变化特点,研究氟对睾丸的毒性影响,选用45只健康性成熟雄性昆明系小鼠,随机分为对照组、低氟组(25 mg/L Na F)、高氟组(100 mg/L Na F),连续饲喂60 d,制作睾丸石蜡组织切片,H.E.染色后,显微镜下观察曲细精管形态变化,通过定量分析方法探讨氟对睾丸的毒性影响。结果表明,与对照组相比,低氟组和高氟组曲细精管半径、面积以及曲细精管管腔半径、面积均无显著性差异(P0.05);高氟组小鼠曲细精管细胞层数和厚度极明显低于对照组(P0.01);形态学观察显示,25 mg/L氟中毒小鼠睾丸生精细胞减少,100 mg/L Na F可导致生精细胞排列紊乱、各级生精细胞减少等。本试验可为氟的雄性生殖毒性研究提供直观、科学的参考。     

苯甲酸钠对小鼠精子畸形率的影响

将50只SPF级成年雄性小鼠随机分为5组:苯甲酸钠低剂量组(0.268g/kg·bw)、中剂量组0.537g/kg·bw)、高剂量组(1.073g/kg·bw)、环磷酰胺阳性对照组(50mg/kg·bw)和生理盐水空白对照组,采用一次性经口灌胃染毒,1次/d,连续5d。于首次染毒后的第35天颈椎脱臼将小鼠处死,取附睾制片,观察小鼠精子形态,对小鼠精子畸形率进行统计分析。结果表明,苯甲酸钠染毒各组小鼠精子畸形率普遍高于空白对照组,差别有统计学意义(P<0.01),且随着染毒剂量的增加,精子畸形率也有相应升高,呈现出一定的剂量-反应关系。     

玉米赤霉烯酮与脱氧雪腐镰刀菌烯醇联合染毒对雄性小鼠生殖毒性的影响及茶多酚的缓解作用

本试验旨在研究茶多酚对玉米赤霉烯酮(ZEN)与脱氧雪腐镰刀菌烯醇(DON)联合染毒小鼠生殖毒性损伤的缓解作用。选取8周龄雄性昆明小鼠75只[体重(40±5)g],随机分成5组,每组分别灌服生理盐水(对照组)、200 mg/kg茶多酚(Ⅰ组)、1 mg/kg ZEN+0.5 mg/kg DON(Ⅱ组)、100 mg/kg茶多酚+1 mg/kg ZEN+0.5 mg/kg DON(Ⅲ组)和200 mg/kg茶多酚+1 mg/kg ZEN+0.5 mg/kg DON(Ⅳ组),每组15只。试验期28 d。在试验第1、14、28天,从每组随机抽取5只小鼠称重采样,检测小鼠精子质量、睾丸脏器系数、睾丸组织标志酶活性、血清性激素含量及睾丸组织氧化与抗氧化指标,并制作睾丸组织切片。结果表明:1)与对照组相比,Ⅱ组小鼠睾丸脏器系数、精子活率及数量、血清性激素含量显著降低(P<0.05),睾丸组织碱性磷酸酶(AKP)、乳酸脱氢酶(LDH)、γ-谷氨酰转移酶(γ-GGT)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)活性显著降低(P<0.05);精子畸形率、血清丙二醛(MAD)含量显著上升(P<0.05)。添加茶多酚后,小鼠的睾丸脏器系数、精子活率及数量、血清性激素含量、睾丸组织AKP、LDH、γ-GGT、CAT、SOD及GSH-Px活性显著上升(P<0.05),精子畸形率、血清MAD含量显著降低(P<0.05)。随着时间的推移,与第1天相比,Ⅰ组小鼠睾丸组织GSH-Px活性显著升高(P<0.05),Ⅱ组小鼠睾丸脏器系数、精子质量、睾丸组织标志酶活性、血清性激素含量、睾丸组织氧化与抗氧化指标均发生显著变化(P<0.05),Ⅲ组小鼠睾丸组织CAT及GSH-Px活性显著降低(P<0.05),Ⅳ组小鼠睾丸组织CAT活性显著降低(P<0.05)。2)睾丸组织病理切片显示染毒组小鼠睾丸生精小管基膜不完整,细胞排列紊乱,数量减少,体积缩小,细胞之间空泡化,管腔内精子数量减少,经茶多酚保护后得到改善。综上,茶多酚能有效改善ZEN与DON联合染毒小鼠精子质量与睾丸组织内标志酶活性,提高血清性激素含量及睾丸组织抗氧化功能,显著降低生殖毒性。     

增精宝对环磷酰胺致小鼠生精障碍保护作用的研究

为评价增精宝对雄性动物生精作用的影响,采用环磷酰胺致小鼠生精障碍为动物模型,考察不同剂量增精宝对环磷酰胺致小鼠生精障碍的保护作用。结果显示,高剂量组对小鼠的附睾、睾丸、储精囊和前列腺均有较显著的促进作用;低剂量组精子总密度和精子活率均差异显著(P<0.05);高剂量组精子活率差异显著(P<0.05);对小鼠睾丸各倍体生精细胞比例的影响,高剂量组小鼠睾丸生精小管较为饱满,管腔内存在大量精子,生精上皮较厚,可见各级生精细胞排列整齐,界膜明显,睾丸间质细胞排列整齐。结论表明,增精宝两种剂量均能够显著促进环磷酰胺导致小鼠生精障碍的保护作用,高剂量组对雄性小鼠的作用尤为显著。     

脱氢表雄酮对雄性小鼠睾丸组织发育的毒性作用

昆明种雄性小鼠32只,随机分为4组。脱氢表雄酮（DHEA）日摄取量分别为0（对照组）、20、40和60mg／kg,试验期52d。各组分别在42日龄、72日龄处死小鼠,分离睾丸,制作石蜡切片,HE染色,显微观察、摄影。结果显示,与对照组比较,42日龄时中、高剂量组的曲精小管断面变形,各时期的生精细胞和成熟精子数量均减少;高剂量组曲精小管内生精细胞层次基本消失,出现较大腔隙,间质发生不同程度的变化。72日龄时中、高剂量组的曲精小管断面变形。基膜变薄伴有断裂和间质受损增加。中剂量组出现畸形精子,高剂量组曲精小管中央空白区域增大,缺乏成熟精子。以上结果表明,中、高剂量的DHEA对生长期小鼠的睾丸组织结构发育有明显的毒性作用。     

Effects of exposure to genistein during pubertal development on the reproductive system of male mice

Genistein, a soybean-originated isoflavone, is widely consumed by humans for putative beneficial health effects but its estrogenic activity may affect adversely the development of the male reproductive system. Twenty-one days old ICR mice weaned from dams fed with a casein-based AIN-76A diet during gestation and lactation were exposed to genistein (2.5 and 5.0 mg/kg/day, p.o.) for 5 weeks. 17beta-Estradiol (7.5 microg/kg/day) and corn oil were used for the positive and negative vehicle controls, respectively. The animals were fed the casein-based AIN-76A diet throughout the experiment. There were no significant differences in body weights of mice between the genistein groups and the negative control group. No significant differences in relative reproductive organ weights were found among all experimental groups. Sperm counts in epididymis and testes were slightly decreased in the genistein-exposed groups compared with control group. Sperm motile characteristics in genistein-exposed groups were slightly higher than those of the control group. Levels of phospholipid hydroxide glutathione peroxidase mRNA in the testis, epididymis, and prostate of mice exposed to genistein or estradiol were significantly higher than those of the controls (P<0.05). Exposure to genistein caused hyperplasia of Leydig cells in the testis and a slight increase of interstitial fibroblasts in the epididymis, while estradiol treatment caused severe damage to the testis and epididymis. These results suggest that dietary uptake of genistein during the juvenile period may affect male reproductive development, resulting in a slight decrease in sperm count, but with an increase in sperm motion quality.     

Effects of exposure to genistein and estradiol on reproductive development in immature male mice weaned from dams adapted to a soy-based commercial diet

Genistein, a soybean-originated isoflavone, is widely consumed by humans for putative beneficial health effects but its estrogenic activity may adversely affect the development of male reproductive system. Twenty one-day-old ICR mice weaned from dams fed with a soybean-based diet throughout gestation and lactation were exposed by gavage to genistein (2.5 mg/kg b.w./day) or 17beta-estradiol (7.5 microg/kg b.w./day) for five weeks. Corn oil was used as a negative control. The animals were fed with a casein-based AIN-76A diet throughout the experimental periods. There were no significant differences in body and organ weights of mice among experimental groups. No significant differences in sperm counts and sperm motile characteristics were found between control and genistein groups. Treatment of 17beta-estradiol caused a significant decrease in prostate weight and epididymal sperm counts compared to the control (p<0.05). The levels of phospholipid hydroxide glutathione peroxidase in the testis and prostate of mice exposed to genistein or 17beta-estradiol were significantly higher than that of the control mice (p<0.05). 17beta-estradiol treatment caused degeneration and apoptosis of germ cells in the testis, depletion and degeneration in the epididymal epithelium, and hyperplasia of mucosal fold region in the prostate of mice. Genistein treatment did not cause any lesion in the testis, epididymis, and prostate. These results suggest that dietary uptake of genistein during juvenile period may not affect male reproductive development and functions.     

氧化乐果对雄性小鼠睾丸琥珀酸脱氢酶和精液质量的影响

试验旨在研究有机磷农药氧化乐果对雄性小鼠睾丸琥珀酸脱氢酶(SDH)和精液质量的影响作用。将40只健康昆明雄性小鼠随机分为4组,即低、中、高3个染毒组和对照组,其中染毒组分别灌胃1.0、2.0、4.0 mg/kg氧化乐果,对照组灌胃生理盐水,采用等容量灌胃法(0.2 mL/10 g体重),连续灌胃14 d。随着氧化乐果染毒剂量的逐渐增加,小鼠睾丸琥珀酸脱氢酶(SDH)活力、精子密度、存活率均明显降低,与对照组相比差异显著(P＜0.05);小鼠体重、睾丸重、附睾及其脏器系数与对照组相比差异均不显著(P＞0.05)。氧化乐果对小鼠精子密度和精子存活率有一定的损伤作用,对睾丸琥珀酸脱氢酶有明显的抑制作用。     

伊维菌素对雄性大白鼠生育功能的影响

探讨伊维菌素对雄性动物生殖功能的影响,为临床用药安全和动物繁殖提供依据。试验选取成年雄性大白鼠,按照0.17 mg/kg（低剂量组）,3 mg/kg（中剂量组）,5 mg/kg（高剂量组）的剂量分别进行灌胃,对照组以生理盐水代替药物进行灌胃。末次给药后24 h处死大鼠,取睾丸、附睾进行生殖系统毒性检查。试验结果表明：当伊维菌素用药剂量达到3 mg/kg以上就呈现出一定的生殖毒性,睾丸系数、附睾系数下降,精子密度和精子活率下降,但对精子活力并没影响,4个试验组均未显示与剂量的相关性。     

还原型谷胱甘肽对氟中毒小鼠附睾的影响

试验旨在研究还原型谷胱甘肽（reduced glutathione,GSH）对氟中毒小鼠附睾的影响。将50只3周龄ICR雄性小鼠随机分为5组,分别为对照组和4组染氟组。对照组饮用蒸馏水,染氟组分别供给含100 mg/L NaF的蒸馏水,自由饮水30 d。之后在染氟组中选择3组每日分别灌胃200、400和800 mg/（kg·只）GSH,同时各组小鼠再自由饮含氟水30 d。试验结束后,摘取小鼠附睾进行活性氧（ROS）、丙二醛（MDA）、GSH含量及谷胱甘肽过氧化物酶（GPx）、谷胱甘肽硫转移酶（GSTs）和谷胱甘肽还原酶（GR）活性检测,同时制作石蜡切片,HE染色后进行组织形态结构分析。结果显示,与对照组相比,100 mg/L NaF组精子密度、精子活力、GSH含量、GPx和GR活性均显著下降（P<0.05）,而GSH添加组较100 mg/L NaF组均有所升高;100 mg/L NaF组精子畸形率、ROS和MDA含量较对照组均显著升高（P<0.05）,而GSH添加组较100 mg/L NaF组均降低,其中800 mg/kg GSH组效果明显。与对照组相比,100 mg/L NaF组附睾头、附睾体及附睾尾管腔厚度升高,管腔内精子数量下降;GSH添加组附睾头、附睾体及附睾尾的管腔厚度与管腔内精子数量都有所恢复,其中800 mg/kg GSH组效果较好。综上所述,添加GSH后氟中毒小鼠谷胱甘肽抗氧化系统酶活性有所升高,附睾组织形态结构一定程度上有所恢复,对附睾有一定的改善作用。     

Spermatogenetic disorders in adult rats exposed to tributyltin chloride during puberty

Adverse effects of tributyltin (TBT) chloride were investigated on the reproductive system in male adult rats as exposed during puberty. Fifty Sprague-Dawley rats at the age of 35 days were assigned to five different groups: negative control receiving vehicle, methyltestosterone (10 mg/kg B.W.), and TBT chloride treatments (5, 10, and 20 mg/kg B.W.). Animals were treated by oral gavage for ten consecutive days and sacrificed at 5 weeks after final treatment. The treatment of TBT chloride at the high dose of 20 mg/kg B.W. significantly decreased homogenization-resistant testicular sperm counts (p<0.05). The TBT chloride treatment at the doses of 10 and 20 mg/kg B.W. also significantly decreased caudal epididymal sperm counts (p<0.01). Some of motion kinematic parameters (motility, mean angular displacement, lateral head displacement, and dance) of sperms retrieved from vasa deference were significantly decreased in rats treated with the TBT chloride at the dose of 20 mg/kg B.W. (p<0.05). These results provide a further evidence that an exposure to TBT chloride during pubertal period in male rats produces spermatogenic disorders characterized by decreasing testicular and epididymal sperm counts and some motion parameters of sperms in the vasa deference.     

Exposure to 3-methyl-4-nitrophenol affects testicular morphology and induces spermatogenic cell apoptosis in immature male rats

The pollutant 3-methyl-4-nitrophenol (PNMC), a major component of diesel exhaust particles, can cause many adverse health problems. In the present study, we investigated the effects of PNMC on the testes of Sprague Dawley rats treated subcutaneously for 5 days with different doses of PNMC (1, 10 or 100 mg/kg). Exposure to PNMC caused a significant decrease in the plasma testosterone levels and in the absolute and relative weights of the testes. Severe histological lesions were observed in the testes of PNMC-treated animals. The ratio of the epithelial height to the seminiferous tubule diameter increased markedly in the PNMC-treated rats compared with the corresponding controls. In addition, PNMC exposure significantly increased the number of apoptotic spermatogenic cells compared with the controls whereas a significant decrease in the ratios of Bcl-2/Bax was observed at the same doses. These results suggest that PNMC exerts its gonadotoxicity in the rat mainly via apoptosis.