三氯异氰尿酸原药对雄性大鼠生殖毒性研究

目的观察三氯异氰尿酸(TCCA)对雄性大鼠生殖系统的影响。方法选取4、12周龄雄性SD大鼠各24只,按周龄、体质量随机分为3组。分别经口灌胃给予0、96.5、193.0 mg/kg剂量水平的受试物10 d。观察大鼠血清生化和Na+、K+、Ca2+、Cl-离子浓度、睾丸曲细精管直径以及睾丸形态和超微结构的变化,评价附睾内精子数量和活动率。结果实验期间未见大鼠死亡;12周龄193.0 mg/kg组大鼠血清乳酸脱氢酶活力升高,血中Ca2+、Cl-降低(P<0.05)。4、12周龄96.5和193.0 mg/kg组大鼠睾丸曲细精管均变细(P<0.01),12周龄大鼠附睾内精子数及精子活动率降低(P<0.01、P<0.05);4周龄大鼠睾丸生精细胞退化增多,12周龄大鼠睾丸出现支持细胞核空泡化、曲细精管生精细胞层次排列紊乱、坏死脱落等病理改变;4、12周龄193.0 mg/kg组大鼠出现精原细胞核膜不完整、核型不规则、染色质浓染等超微结构改变。结论 TCCA能诱导未成年大鼠和成年大鼠的睾丸毒性,表现为睾丸结构的破坏和生精功能的抑制,其毒性作用机制尚待深入研究。     

Restraint stress exacerbates alcohol-induced reproductive toxicity in male rats

Cumulative exposure to multiple stresses may lead to aggravating the toxicity of each stress, qualitatively or quantitatively altering biological responses because of toxicological interaction. In this study, we intended to determine the possible effects of restraint stress on reproductive toxicity due to ethanol usage in male rats. Early pubertal male Wistar rats were subjected to either restraint stress (5 h/day) or alcohol intoxication (2 mg/kg body weight) or both for 60 days. Body weights of control and experimental rats were similar during the 60 days of this study. Testes were harvested, weighed, and prepared for enzyme assays, and cauda epididymides were isolated for the determination of density, motility, and viability of stored spermatozoa. Restraint stress or alcohol treatment significantly reduced testis weight and caused significant reductions in steroidogenesis and spermatogenesis. Mean density, motility, and viability of stored spermatozoa were reduced in experimental rats. Plasma testosterone concentrations in rats subjected to restraint stress or alcohol were decreased compared with those of controls, concomitant with increased concentrations of LH and FSH in experimental rats. These data suggest that sub-chronic exposure to restraint stress or alcohol contribute to reduce testicular and epididymal function in exposed rats. The study also suggests that restraint stress exacerbates alcohol-induced reproductive toxicity in rats.     

慢性不可预知性温和刺激对雄性大鼠生殖系统的影响

目的:研究慢性不可预知性温和刺激对雄性大鼠生殖系统的影响。方法:将SD雄性大鼠随机分为正常对照组和模型组。模型组大鼠每天给予多种低强度刺激,连续28d,构建模型。称量大鼠体重、睾丸和附睾重量。取出附睾计数精子、观察精子活率。HE染色观察大鼠睾丸组织结构。结果:与正常对照组相比,模型大鼠体重、睾丸重量、附睾重量显著减少,精子活率显著下降,a、c级精子所占比例明显下降;组织形态学与对照组相比,模型组大鼠睾丸组织受到明显损伤。结论:慢性不可预知性温和刺激可损伤雄性大鼠的生殖功能。     

长期低铅染毒大鼠脾脏光镜及电镜结构的观察

应用光镜和透射电镜技术观察长期低铅染毒后大鼠脾脏的结构改变。光镜下可见,脾脏边缘带相对面积减小,脾小结个为数明显减少。电镜观察可见,脾脏免疫细胞出现不同程度的线粒体肿胀、空泡化或固缩;内质网肿胀、脱颗粒。上述变化均随着铅染毒剂量的增加而加重。结果提示,长期低铅染毒可导致脾脏免疫细胞的损伤。     

乙酸铅暴露对雄性大鼠心血管系统的炎性作用

目的探讨乙酸铅经口暴露对雄性大鼠心血管系统炎性指标的影响。方法将40只健康成年SPF级雄性Wistar大鼠随机分为4组,分别为对照(0 mg/kg)组和90、180、360 mg/kg乙酸铅染毒组,每组10只。采用经口灌胃方式进行染毒,染毒容量为10 ml/kg,每天1次,每周连续染毒5 d,持续染毒60 d。测定大鼠血铅、血浆内皮素1(ET-1)、血清细胞间黏附分子-1(ICAM-1)及血清和心脏组织匀浆中白介素-1β(IL-1β)、白介素-6(IL-6)、白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)的表达水平;同时,观察大鼠心脏和胸主动脉组织病理学变化。结果与对照组相比,各剂量乙酸铅染毒组大鼠的血铅水平均升高,差异有统计学意义(P0.05);且随着乙酸铅染毒剂量的升高,大鼠的血铅水平呈上升趋势。与对照组相比,360 mg/kg乙酸铅染毒组大鼠血浆ET-1和血清ICAM-1水平均升高,差异有统计学意义(P0.05);且随着乙酸铅染毒剂量的升高,大鼠的血浆ET-1和血清ICAM-1水平均呈上升趋势。与对照组比较,360 mg/kg乙酸铅染毒组大鼠血清中IL-β、IL-8和TNF-α的水平及180 mg/kg乙酸铅染毒组大鼠血清中IL-8和TNF-α水平均升高,差异有统计学意义(P0.05);而各剂量乙酸铅染毒组大鼠血清中IL-6的水平均无明显改变。且随着乙酸铅染毒剂量的升高,大鼠血清中IL-β、IL-6、IL-8和TNF-α的水平均呈上升趋势。与对照组比较,仅360 mg/kg乙酸铅染毒组大鼠心脏组织匀浆中IL-8水平升高,差异有统计学意义(P0.05);而各剂量乙酸铅染毒组大鼠心脏组织匀浆中IL-1β、IL-6、TNF-α的水平均无明显改变。且随着乙酸铅染毒剂量的升高,大鼠血清中IL-8和TNF-α的水平均呈上升趋势,而IL-β、IL-6的水平均呈先上升后下降的趋势。大鼠血清和心脏组织匀浆中IL-8和TNF-α水平均呈正相关(均P0.05),而两者IL-1β和IL-6水平间无统计学相关性(P0.05)。病理学检查可见,180、360 mg/kg乙酸铅染毒组心脏组织出现炎性细胞浸润,360 mg/kg乙酸铅染毒组血管组织外膜出现炎性细胞浸润。结论乙酸铅对大鼠心血管系统具有一定的炎性损伤作用。     

铅和乙醇对雄性大鼠生殖系统的联合毒作用

采用整体动物实验方法,观察铅和乙醇对大鼠精子数量和质量,以及血中性激素水平的联合作用。结果显示,精子计数联合组比铅和乙醇单独作用组显著减少;精子活动度分析联合组与其他各组比较显著下降。铅和乙醇单独作用均可使雄性大鼠血清睾酮(T)升高,促黄体生成激素(LH)下降,联合作用使T降低,LH较单独作用组升高。提示铅和乙醇联合染毒对雄性大鼠生殖毒性影响可能具有增毒作用。     

铅、锰染毒后大鼠精子形态的改变

目的观察不同剂量的铅、锰染毒后,大鼠精子数量及形态的改变。方法成年大鼠经口给予低剂量醋酸铅(0.05%、0.1%和0.2%)染毒,连续染毒8周。氯化锰51、5和45 mg/kg连续染毒6周。结果光学显微镜下观察发现,染毒后大鼠成熟精子数量明显减小,头部畸形和尾部畸形的精子数量随铅、锰染毒剂量的增加而增加。结论一定剂量的铅、锰可影响雄性大鼠生殖功能。     

乙醇的雄性生殖毒性

40只健康雄性成年SD大鼠随机分为4组,各组每日分别灌胃给予乙醇0、2.7、4.5、7.5g/kg,连续13周。测定各组大鼠血清睾酮(T)、促黄体生成素(LH)、促卵泡刺激素(FSH)含量,光、电镜观察睾丸组织的形态学改变,同时测定睾丸线粒体中丙二醛(MDA)的含量。结果显示各乙醇组动物血清T水平较对照组明显降低(P<0.01),血清LH、FSH含量亦明显降低(P<0.05)。睾丸的组织病理学观察显示染毒动物睾丸生精细胞核固缩、变性,曲细精管腔中脱落细胞增多;超微结构显示睾丸生精上皮结构破坏,支持细胞和各级生精细胞均有退化变性,且损伤程度与乙醇剂量明显相关。乙醇4.5、7.5g/kg组动物睾丸线粒体MDA含量明显高于对照组(P<0.05)。说明乙醇既可以直接作用于睾丸,引起生精细胞损伤和睾丸类固醇合成抑制,还可使下丘脑垂体轴生殖内分泌功能受损。脂质过氧化可能是乙醇致睾丸损伤的机制之一。     

染铅大鼠子代脑铅含量及对神经行为功能的影响

为探讨染铅大鼠子代脑铅含量及对神经行为功能的影响, 将Ｗｉｓｔａｒ 大鼠６０ 只随机分成高、中、低剂量和对照组, 进行醋酸铅饮水染毒实验。结果显示： 染毒各组大鼠子代脑铅含量均高于对照组, 呈随染毒剂量增加而增高的趋势。悬崖回避试验阳性率较对照组低且有显著差异（ Ｐ＜ ０－０１） ; 学习、记忆功能染毒各组均比对照组时间明显延长（ Ｐ＜ ０－０１） 。提示： 铅通过母体对子代造成脑铅的蓄积, 导致早期行为发育和学习记忆功能的损害。     

洗洁精对雄性大鼠生殖功能的影响

目的:评价家庭日常生活用洗洁精对雄性大鼠的生殖毒理。方法:将2种市售民用洗洁精(LB、BM)以1/万、1‰、1%3个剂量溶于饮水中,10周龄SD雄性大鼠自由摄取连续100d,以饮用正常水大鼠为对照组,进行比较分析。结果:LB和BM洗洁精3个剂量下,对动物生长发育和体重均无明显影响,对生殖系统的附睾和会阴复合体组织重量无明显影响,在1%剂量下睾丸组织明显减轻(P(0.05),比对照组分别下降12.3%和11.7%。2种洗洁精在3个剂量下的储精囊和前列腺均水肿,重量明显增加,表现出明显的剂量效应关系;精子功能分析中运动性、精子数目、精子形态等指标,均明显下降(P(0.05～0.0001),且表现出明显的剂量效应关系;睾丸组织形态分析显示LB在1%剂量下生精功能受到明显抑制和阻断。实验结果表明生殖毒性LB强于BM。结论:雄鼠在大剂量长期摄入LB和BM洗洁精可产生明显的生殖毒性作用。     

Fas凋亡相关基因在双酚A诱导成年雄性大鼠生殖毒性中的作用

目的探讨Fas凋亡途径相关基因在双酚A（BPA）诱导成年雄性大鼠生殖毒性中的作用。方法 40只成年Wistar雄性大鼠随机等分为4组：溶媒对照组（玉米油）和低、中、高剂量BPA喂养组（10、50、250μg/kg·bw/d）。大鼠在喂养35周后处死,称量睾丸、附睾、肝、肾及心脏重量并计算脏器系数;观察精子形态及计算精子畸形率。采用荧光实时定量PCR（RT-PCR）测定睾丸组织中自杀相关因子（Fas/Fas-L）、天冬氨酸特异酶切的半胱氨酸蛋白酶3（Caspase-3）mRNA含量。结果高剂量BPA喂养大鼠精子畸形率以及睾丸组织中Fas-L mRNA及Caspase-3 mRNA含量高于对照组,差异均有显著性（P!0.05）。结论在BPA的人体每日容许摄入量水平下（TDI=50μg/kg·bw/d）,未观察到生殖细胞损伤的效应;但长期经膳食暴露高剂量BPA（250μg/kg·bw/d）可致成年雄性大鼠生殖细胞损伤,其机制可能与BPA上调Fas-L mRNA和Caspase-3的表达有关。     

氯化汞对雄性大鼠生殖毒性的研究

目的探讨无机汞对雄性生殖系统的损伤机制.方法雄性Wistar大鼠随机分3组,分别隔日皮下注射剂量为1.0 mg/kg、3.0 mg/kg的HgCl2及生理盐水,连续21 d.观测睾丸及附睾脏器系数、生精细胞凋亡情况、脂质过氧化物(LPO)、一氧化氮(NO)含量以及染毒前后血清睾酮水平.结果(1)3.0 mg/kg HgCl2染毒组大鼠睾丸及附睾脏器系数分别为0.74、0 26,均低于对照组(睾丸0 89,附睾0.30),差异有显著性(P＜0.05).(2)1.0 mg/kg及3.0 mg/kg HgCl2染毒组染毒后睾酮水平分别为0 93、0 69 nmol/L,均低于染毒前(5.01、3.02 nmol/L)及对照组(5 49 nmol/L),差异有显著性(P＜0.05).(3)1.0 mg/kg和3 0 mg/kg HgCl2染毒组LPO分别为17.34、21.59 μmol/ml;NO分别为62 91、73.09μmol/ml,均高于对照组(LPO 10.32 μmol/ml,NO 49 86μmol/ml),差异有显著性(P＜0.05).(4)3 mg/kg HgCl2染毒组平均曲细精管面积为56 159.01μm2/个,低于对照组(1 222 274.39μm2/个),差异有显著性(P＜0.05).1.0 mg/kg HgCl2染毒组平均每个曲细精管的凋亡细胞数为114.20个,高于对照组(39 02个),差异有显著性(P＜0.05).1、3.0 mg/kg HgCl2染毒组单位曲细精管面积内凋亡细胞数分别为1 06×10-3个/μm2、1.04 × 10-3个/μm2,均高于对照组(0.33个),差异有显著性(P＜0.05).结论HgCl2可诱发大鼠生精细胞凋亡,可能是对雄激素缺失信号的应答;睾丸组织处于氧化应激状态和生精细胞凋亡增加、睾酮水平下降可能有一定关系.     

2-乙氧基乙醇急性染毒大鼠血清和睾丸某些抗氧化指标的变化

目的 观察2-乙氧基乙醇(2-Ethoxythanol,EE)急性染毒对SD大鼠血清和睾丸某些抗氧化指标的变化，探讨EE致睾丸损伤的可能机制。方法 选择健康雄性SD大鼠，体重180-220g。随机分为对照组、EE800、1600和3200mg／kg组4组，每组24只。采取一次性灌胃染毒。于灌胃后12、24、48和72h，将各组动物随机处死6只，留取动物血液、睾丸，制备血清和睾丸匀浆，测定血清和睾丸匀浆脂质过氧化物(LPO)水平、超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性，以及血清铜蓝蛋白(CP)活性。结果 与对照组比较，各染毒组睾／体比明显下降(P＜0.05)，睾丸匀浆LPO水平和血清CP活性增高。染毒12、24h，血清CAT、睾丸匀浆CAT和SOD活性增高，而染毒48、72h后，血清CAT、睾丸匀浆CAT和SOD活性显著降低(P＜0.05)。EE各染毒组血清LPO水平和SOD活性变化不明显。结论 推测EE毒作用的靶器官可能是睾丸，睾丸抗氧化功能的改变是EE致睾丸毒性的可能机制。     

Evaluation of antifertility potential of Brahmi in male mouse

Background

The purpose of the present study was to evaluate the effect of Bacopa monnieri (Brahmi) on fertility of male laboratory mouse.

Study Design

Mice of the Parkes (P) strain were orally administered Brahmi (250 mg/kg body weight/day, for 28 and 56 days), and effect of the treatment on reproductive organs and fertility was investigated. Recovery and toxicological studies were also carried out.

Results

The treatment caused reduction in motility, viability, morphology, and number of spermatozoa in cauda epididymidis. Histologically, testes in mice treated with the plant extract showed alterations in the seminiferous tubules, and the alterations included intraepithelial vacuolation, loosening of germinal epithelium, exfoliation of germ cells and occurrence of giant cells. In severe cases, the tubules were lined by only Sertoli cells or Sertoli cells, spermatogonia and spermatocytes. Significant reductions were also noted in height of the germinal epithelium and diameter of the seminiferous tubules in Brahmi-treated mice compared to controls. Epididymis in treated males showed slight alterations in histological appearance. The treatment had no effect on levels of testosterone, alanine aminotransferase, aspartate aminotransferase and creatinine in blood serum, hematological parameters and on liver and kidney histoarchitecture. In Brahmi-treated males, libido remained unaffected, but fertility was notably suppressed. The alterations caused in the above reproductive endpoints by the plant extract were reversible, and by 56 days of treatment withdrawal, the parameters recovered to control levels.

Conclusions

The results in P mice thus suggest that Brahmi treatment causes reversible suppression of spermatogenesis and fertility, without producing apparent toxic effects.     

Evaluation of reversible contraceptive activities of Cuminum cyminum in male albino rats

Background

The aim of the present study was to evaluate the contraceptive efficacy of Cuminum cyminum (jeera) seeds in male albino rats.

Study Design

C. cyminum methanol extract (CcMtE) at dose levels of 100 and 200 mg/rat/day was orally administered to male rats for 60 days. The effect of the treatment on reproductive organs and fertility was investigated. Recovery and toxicity studies were also carried out.

Results

C. cyminum methanol extract fed to male rats for 60 days did not cause any alterations in the body weight, whereas the weight of testes, epididymides, seminal vesicles and ventral prostate were significantly reduced (p≤.001). Animals treated with CcMtE showed a marked reduction in sperm density in the cauda epididymis and testes and sperm motility in the cauda epididymis. Reduction in fertility was 69.0% and 76.0% in 100 and 200 mg/rat/day dose levels, respectively. The circulatory hormones were also reduced significantly. Testicular biochemical analysis of protein, sialic acid, glycogen, ascorbic acid and fructose indicated a marked decline, whereas testicular cholesterol content was significantly increased, which showed altered biochemistry of the reproductive organs. After CcMtE treatment, significant decreases (p≤.001) were observed in the number of testicular cells (i.e., spermatogonia, primary spermatocytes [preleptotene and pachytene], secondary spermatocytes and round spermatids); nonsignificant change was observed in the Sertoli cell count. The treatment had no effect on levels of serum protein, cholesterol, bilirubin, glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), blood urea and hematological indices.

Conclusions

The present study shows that C. cyminum treatment resulted in the inhibition of spermatogenesis and fertility without producing apparent toxic effects.     

醋酸铅致大鼠血淋巴细胞DNA损伤的研究

目的 研究醋酸铅致淋巴细胞的DNA损伤情况,了解铅的遗传毒性.方法 用不同浓度的醋酸铅分别对大鼠进行腹腔注射体内染毒24h及取正常大鼠血淋巴细胞进行体外染毒1 h,然后,采用单细胞凝胶电泳技术分别测定各血淋巴细胞DNA损伤程度.结果 体内实验组见淋巴细胞边缘不光滑,毛糙,体外实验各处理组都有明显彗尾形成,体内外实验各染毒组彗星出现率、DNA迁移长度均明显高于阴性对照组.结论 醋酸铅对大鼠淋巴细胞DNA具有一定的损伤作用.     

Developmental Changes in Testosterone Production by the Rat Testis in Vitro During Late Fetal Life

The age-related evolution of the in vitro capacity of the rat testis to produce testosterone and to respond to luteinizing hormone (LH) during 3 and 24 h incubation was studied from day 18.5 to day 21.5 of fetal life. Basal testosterone production by testes from 18.5-day-old fetuses was significantly higher than production by testes from 20.5- and 21.5-day-old fetuses when secretion was expressed as either per testis or per microgram of testicular protein. When maximal LH-stimulated testosterone secretion was expressed on a per testis basis, it was significantly lower for day 18.5 testes than for day 20.5 and 21.5 testes. However, when it was expressed on the basis of testicular protein content, it decreased significantly between days 18.5 and 20.5. Basal and LH-stimulated secretions displayed the same time-related decrease throughout 24 h of incubation for the three ages studied. The dose-response curves for LH showed that the sensitivity was similar for day 18.5 and 20.5 testes (ED₅₀ = 10 vs. 14 ng/mL, respectively). These results showed an age-related decrease in testicular steroidogenic capacity without a change in the coupling efficiency of LH receptor to testosterone production during late fetal life in rats.     

大剂量PN诱发雄性大鼠性腺损伤的实验研究

雄性成年ＳＤ大鼠随机分为５组,分别于右侧睾丸内注射生理盐水、７０％甘油和ＰＮ。７天后重复１次。第１５天取样,进行精子分析、睾丸及主要副性腺测量和组织病理学观察。结果显示,睾丸内注射ＰＮ引起了性腺、副性腺萎缩,精子性状改变以及组织病理学改变。推测可能是：（１）ＰＮ或ＰＬ与ＰＬＰ竞争结合部位和抑制ＰＬＰ的合成,干扰细胞代谢;（２）通过转化形式或直接作用于Ｓｅｒｔｏｌｉ细胞,损害细胞骨架。（３）直接透过血睾屏障而损害管腔小室内的生精细胞     

孕期染毒壬基酚对子代雄性胎鼠睾丸发育和胰岛素样因子-3表达的影响

目的 探讨母体染毒壬基酚(NP)对雄性胎鼠生殖发育的影响,并检测胎鼠睾丸组织中胰岛素样因子-3(Insl-3)的mRNA以及蛋白的表达.方法 将已受孕的SD雌性大鼠随机分为对照组、5、40、100、200 mg/kg NP染毒组(每组20只),染毒组大鼠于孕14～19 d以NP灌胃染毒,对照组大鼠用纯花生油灌胃第19天处死孕鼠后检测孕鼠和雄性胎鼠生殖发育相关指标;反转录-聚合酶链式反应(RT-PCR)和蛋白免疫印迹技术(Western blot)检测其睾丸组织中Insl-3的mRNA及蛋白的表达.结果 与对照组比较,40、100、200 mg/kg组胎鼠体重和睾丸系数明显降低,差异有统计学意义(P＜0.05或P＜0.01).与对照组比较,40、100、200 mg/kg组的睾丸下降程度降低,差异有统计学意义(P＜0.05).与对照组比较,40、100、200 mg/kg组Insl-3mRNA及蛋白表达降低,差异有统计学意义(P＜0.05或P＜0.01).结论 孕期暴露NP可致其雄性胎鼠睾丸发育不良,下降不全,并且可下调Insl-3的表达.     

铅染毒小鼠睾丸组织和生化变化及某中药对其治疗效果观察

目的 探讨化学毒物铅对小鼠生理、生化指标、睾丸组织形态学的影响以及某中药口服液对其治疗效果的观察.方法 实验前给小鼠称体重、编号、测定血红蛋白(Hb).用乙酸铅水溶液给小鼠作腹腔注射,染毒2周后,将染毒小鼠分成3组:模型组、治疗1组和治疗2组,不染毒的小鼠为正常对照组.分组后进行治疗,治疗组用某中药口服液灌胃,1次/d...