巴戟天萃取物对环磷酰胺诱导睾丸损伤模型大鼠的影响

目的:观察巴戟天萃取物(MO)对环磷酰胺(CTX)诱导睾丸损伤模型大鼠的影响及可能机制。方法:雄性SD大鼠,随机分为空白对照组、CTX+NS组、CTX+MO不同剂量组。以环磷酰胺构建大鼠睾丸损伤动物模型;巴戟天水提物用乙酸乙酯萃取后取水层成分给予每组治疗组灌胃。观察各组大鼠睾丸微细结构,比较卵泡刺激素受体(FSHR)蛋白及mRNA表达的差异。结果:光镜下观察可见CTX+NS组明显病理改变,各治疗组的睾丸生精小管的结构明显改善,FSHR mRNA和FSHR蛋白表达明显高于CTX+NS组,差异具有统计学意义。结论:巴戟天萃取物对环磷酰胺诱导睾丸损伤模型大鼠具有修复作用,可能与巴戟天萃取物能促进睾丸FSHR高表达有关,并以30g·kg~(-1)·d~(-1)浓度效果最佳。     

环磷酰胺对大鼠睾丸C-Kit表达的影响

目的探讨抗肿瘤药物环磷酰胺对大鼠睾丸中分化型精原干细胞的特异性表面标志物c-kit表达的影响。方法 20只8周龄大鼠随机分为对照组和实验组,每组10只;实验组每天腹腔注射环磷酰胺50 mg·kg-1·d-1,注射3 d,对照组腹腔注射0.9%的生理盐水;用药4周后取双侧睾丸,应用HE染色、免疫组织化学技术和Western blot技术,观察睾丸组织学变化、c-kit在睾丸中的分布情况及表达变化。结果 1实验组大鼠睾丸中生精上皮呈有序性排列,但生精上皮层数减少,管腔中精子数量减少。2免疫组化结果显示c-kit在睾丸的生精细胞中均匀表达,但实验组的c-kit表达明显下降。结论环磷酰胺对大鼠生精功能的损害可能和通过下调c-kit的表达有关。     

巴戟天对微波损伤的雄鼠睾丸生精功能的影响

目的探讨微波辐射对雄鼠的生精功能的影响及巴戟天对损伤后雄鼠的生精功能的修复作用。方法 24只SD大鼠随机均分为空白组、辐射组和给药组。用对讲机微波辐照辐射组和给药组;给药组在辐射后巴戟天灌胃。三组均在相同条件下饲养。1周后观察各组雄鼠的睾丸指数、精子活性及精子密度等指标的变化。结果辐射组相对空白组睾丸指数、精子活性以及精子密度降低(P0.05),附睾指数变化没有统计学意义(P0.05);给药组相对辐射组附睾指数、睾丸指数、精子活性、精子活动率均增高(P0.05);而给药组相对空白组睾丸指数、精子活性以及精子密度增高(P0.05),附睾指数变化没有统计学意义(P0.05)。结论巴戟天对微波损伤的雄鼠睾丸生精功能有修复作用。     

淋巴管阻断对大鼠睾丸生精上皮的影响

用外科手术的方法将正常成年ＳＤ大鼠双侧睾丸淋巴管阻断后，观察术后不同时间睾丸生精上皮的组织学变化。术后２天出现生精上皮的形态改变，生精细胞排列松散，部分精子细胞脱落，聚集在管腔部位。术后６天多数曲细精管的精子细胞全部脱落。假手术对照组，睾丸生精上皮具有正常的形态结构。     

磁水对家兔睾丸生精功能的影响

用磁水饲养家兔，经3和6个月的观察，实验组动物 精液量、精子数、精子存活率，附睾尾中精子数以及每100mg附睾尾组织的精子烽，曲细精管直径及曲细精管生精细胞数都比对照组显著增加；精子畸形率比对照组明显减少，光镜及扫描电镜观察显示实验组精母细胞、精子细胞、精子数量都比对照组显著增加，表明饮用磁水能促进生精功能。     

环磷酰胺对大鼠睾丸和附睾的影响

目的探讨抗肿瘤药物所致大鼠睾丸和附睾损害,为寻找男性不育症中药治疗的研究提供理论依据。方法选用16只15周龄SD大鼠随机分为对照组和实验组,每组8只;实验组腹腔注射环磷酰胺20mg·kg-1·d-1,连续5d,用药2个月后,应用HE染色法研究大鼠睾丸、附睾远期组织学变化,用原位缺口末端标记法(TUNEL方法)检测生精细胞凋亡。结果实验组大鼠体重、睾丸和附睾重量均显著减轻(P<0.01),睾丸生精小管直径缩小、间距增宽、生精上皮变薄、生精细胞层次和数量减少、生精小管腔多未见精子形成,实验组睾丸生精小管直径、面积、生精上皮细胞数、均显著低于对照组(P<0.01);实验组生精细胞凋亡增多,与对照组比较,生精细胞显著凋亡(P<0.01);附睾管管腔内腔内精子稀少,含有大量脱落细胞,管壁变薄。结论环磷酰胺对大鼠睾丸、附睾远期损害明显,促进生精细胞凋亡。     

冬虫夏草对抗环磷酰胺致小鼠生精障碍的作用

目的冬虫夏草(cordyceps)具有清除自由基,抗氧化的作用。本实验研究冬虫夏草对抗环磷酰胺致小鼠生精功能损害的作用。方法随机将小鼠分成3组:正常对照组、模型对照组、冬虫夏草治疗组。模型对照组和冬虫夏草治疗组小鼠按照40mg/kg.d腹腔注射环磷酰胺(cp)连续5天,同时冬虫夏草治疗组小鼠每天给予冬虫夏草混悬液300mg/kg.d灌胃;正常对照组小鼠及模型对照组灌胃等量生理盐水。各组均连续处理28天。末次给药24h后,处死全部小鼠,观察小鼠附睾精子密度,活力,活动率,睾丸组织匀浆后测定超氧化物歧化酶(superoxide d ismutas,SOD),谷胱甘肽过氧化物酶(glutath ione Peroxidase,GSH-Px),丙二醛(m alon iealdehvde,MDA)含量。结果与模型对照组相比,治疗组精子密度,活力,活动率,睾丸组织SOD与GSH-Px含量等均明显升高(P<0.05,P<0.01),精子畸形率和MDA含量有明显降低(P<0.05,P<0.01)。结论冬虫夏草可对抗环磷酰胺所致的小鼠生精功能的损害,其作用机制可能与冬虫夏草可清除氧自由基,抗氧化作用有关。     

巴戟天多糖对精索静脉曲张大鼠睾丸修复作用的蛋白质组学分析

目的：从蛋白质组学层面探讨精索静脉曲张（VC）的病理机制及巴戟天多糖（MOP）对VC大鼠睾丸修复 作用的机制。方法：健康雄性SD大鼠随机分为假手术组、VC模型组、VC+MOP给药组。采用串联质谱标签法 对大鼠左侧睾丸组织进行蛋白质组学分析,筛选差异表达蛋白（ 差异倍数>25%）,并进行生物信息学分析。结 果：VC模型组和假手术组相比差异蛋白总数为46 个,VC+MOP给药组与VC模型组相比差异蛋白总数为21 个, VC+MOP给药组与假手术相比差异蛋白总数为5 个。差异表达蛋白层次聚类分析结果显示,VC+MOP给药组与 假手术组之间蛋白表达相似,而VC模型组与其他2 个组差异较大。基因本体（GO）和日本京都基因和基因组百 科全书（KEGG）通路富集分析显示,差异蛋白主要富集于核糖体生物合成、细胞凋亡信号、激素水平调节、血 液循环调节、体液免疫应答、蛋白激酶C信号转导、肽激素分泌通路。结论：MOP可使由VC引起的组间睾丸差 异表达蛋白数量减少,逆转由VC引起的蛋白表达水平异常,提示VC的病理机制和MOP的修复作用可能与关键 蛋白的差异表达以及富集的GO和KEGG通路有关。     

砷染毒对大鼠睾丸凋亡诱导因子表达及生精细胞凋亡的影响

目的:探讨砷染毒对大鼠睾丸凋亡诱导因子(AIF)表达及生精细胞凋亡的影响。方法:雄性SD大鼠被随机分为对照组和染砷组。采用自由饮用方式进行连续染毒6个月。采用免疫印迹及实时荧光定量PCR检测AIF表达水平;采用TUNEL法观察生精细胞凋亡的情况。结果:免疫印迹及实时荧光定量PCR结果显示,与对照组比较,染砷组大鼠睾丸内AIF蛋白及mRNA表达水平均显著增高。TUNEL法检测结果显示与对照组比较,染砷组生精上皮凋亡细胞平均灰度值显著增高。结论:砷染毒时AIF介导的非caspase依赖性凋亡途径可能参与了大鼠生精细胞凋亡。     

淫羊藿苷调节雄性大鼠生殖功能

目的:探讨淫羊藿苷在环磷酰胺诱导的大鼠睾丸生精障碍动物模型中的作用,研究淫羊霍苷对睾丸功能的影响.方法:分为空白对照组、阴性对照组、模型组、淫羊藿苷治疗组;H-E染色观察睾丸组织结构变化,TUNEL方法检测睾丸生殖细胞凋亡,放射免疫法检测血清睾酮.结果:睾丸H-E染色切片观察显示模型组睾丸生精小管直径缩小,间距增宽,生精上皮变薄,生殖细胞数量减少,生精小管多未见精子形成,与空白对照组比较其生精小管结构变化显著;淫羊藿苷治疗组与模型组比较生精小管壁增厚,含有精子的生精小管明显增多.睾丸生精小管中生殖细胞凋亡的观察模型组与空白对照组比较其生殖细胞凋亡增多,变化显著;淫羊藿苷治疗组与模型组比较生精小管中生殖细胞凋亡数量明显减少.模型组与空白对照组比较血清睾酮明显降低;淫羊藿苷治疗组与模型组比较其血清睾酮明显增加.结论:淫羊藿苷对环磷酰胺诱导生精障碍的睾丸具有改善睾丸生精小管结构、减少生殖细胞凋亡、促进精子发生和间质细胞分泌睾酮的功能.     

Expression of mucin genes in the human testis and its relationship to spermatogenesis

In this study we investigate the expression pattern of mucin genes in the human testis and evaluate the relationship between the expression of mucin genes and impaired spermatogenesis in the human testis. Thirty human testis tissues were collected from patients undergoing diagnostic testicular biopsy to investigate the cause of infertility. One part of the tissue underwent histological observation, and the other part of the tissue was subjected to semiquantitative RT-PCR of mucin genes, that is, mucin1, 2, 3, 4, and 9. The relative amount of mucin mRNAs was calculated by densitometry using glyceraldehydes-3-phosphate dehydrogenase (GAPDH) as an internal control. The samples were histologically diagnosed as either obstructive azoospermia with normal spermatogenesis (n = 13) or non-obstructive azoospermia with impaired spermatogenesis (n = 17). In the human testis with normal spermatogenesis, mRNA expression of mucin1, 9, 13 and GAPDH were found, but RT-PCR products of mucin 2, 3 and 4 were not detected. In the testis with impaired spermatogenesis, however, RT-PCR product of mucin1 was not found. There was no difference in the other mucin mRNA expression patterns between the testis with either normal or impaired spermatogenesis. To our knowledge, this study is the first that has detected the mRNA of mucin9 and 13 in human testis. This study also shows that mucin1 expression might be closely related to spermatogenesis. Our findings should be substantiated by more direct evidence, such as mucin protein expression and localization.     

巴戟天寡糖单体HexB减轻HUVECs缺氧复氧损伤

目的:研究巴戟天寡糖单体HexB减轻缺氧复氧(H/R)损伤诱导人脐静脉内皮细胞(HUVECs)凋亡的分子机制。方法:HUVECs分别用HexB、内质网应激(ERS)抑制剂4-苯基丁酸(4-PBA)和ERS诱导剂毒胡萝卜素(TG)干预,并将培养的HUVECs分为:对照组、HexB组、H/R组、HexB+H/R组、4-PBA+H/R组、TG组和HexB+TG组。CCK-8法和流式细胞术检测细胞活性及凋亡率,Western blot法检测ERS标志分子葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)、凋亡相关蛋白caspase-12及磷酸化c-Jun氨基末端激酶(p-JNK)的蛋白水平。结果:与对照组比较,H/R组和TG组的细胞凋亡率明显增加,细胞活力明显降低,GRP78、CHOP、caspase-12及p-JNK的蛋白水平上调(P0.05);与H/R组比较,HexB+H/R组及4-PBA+H/R组的细胞凋亡率显著降低,细胞活力升高,GRP78、CHOP、caspase-12及p-JNK的蛋白水平降低(P0.05);与TG组比较,HexB+TG组的细胞凋亡率显著降低,细胞活力升高,GRP78、CHOP、caspase-12及p-JNK的蛋白水平降低(P0.05);HexB+H/R组与4-PBA+H/R组比较,细胞凋亡率、细胞活力及GRP78、CHOP、caspase-12、p-JNK蛋白水平的差异无统计学显著性。结论:通过抑制内质网应激,HexB可以减轻H/R诱导的HUVECs凋亡,其机制可能与下调GRP78、CHOP、p-JNK及caspase-12的水平有关。     

Neonatal exposure of male rats to estradiol benzoate causes rete testis dilation and backflow impairment of spermatogenesis

Estrogens administered to perinatal rodents cause spermatogenesis impairment; this study was undertaken to determine the mechanisms by which estrogens exert this effect. Neonatal male Wistar rats received estradiol benzoate (either 0.5 mg/5g BW or 1 mg/5g BW) and were killed at days 10, 22, 33, 45, and 60. Controls received vehicle. In tubule cross-sections of transverse sections of the right testes, 1) tubular diameter (TD) and seminiferous epithelium height (SEH) were measured, 2) normal and impaired spermatogenesis were classified in terms of the most advanced germ cell type present, including tubules lined by Sertoli cells only. A significant dose-dependent rise in the tubule percentage lined by Sertoli cells only at day 60 reflected spermatogenesis impairment. This was evidenced by the presence of multinucleated germ cells in a thin epithelium and sloughed into an enlarged tubular lumen, which was reflected in a significant dose-dependent increase in TD/SEH values from day 22 onward. TD was significantly greater and SEH significantly lower in tubular segments located at the cranial than the caudal halves of rat testes treated with the high (days 22, 33, and 60) and the low dose (day 33). This indicated distension in cranial tubular segments, perhaps due to the fact that these segments were the closest to the dilated rete testis. Consequently, they showed the highest TD/SEH values and the most regressive features of spermatogenesis (tubules lined by Sertoli cells only). In contrast, caudal segments in rat testes treated with the low dose showing TD/SEH values similar to controls displayed a delayed maturation of spermatogenesis coinciding with the late appearance of mature Leydig cells. Anat. Rec. 252:17–33, 1998. © 1998 Wiley-Liss, Inc.     

Role of estrogen in spermatogenesis in initial phase males of the three‐spot wrasse (Halichoeres trimaculatus): Effect of aromatase inhibitor on the testis

The three‐spot wrasse, Halichoeres trimaculatus, is a protogynous hermaphrodite. Under appropriate social conditions, female fish can become male. Previous studies indicated that estrogens are important regulators of sex change in this fish. However, the role of estrogen in the male is not known. To clarify the involvement of estrogen in spermatogenesis in hermaphrodite fish, we treated initial phase (IP) males for 10 weeks with exemestane, an aromatase inhibitor (AI), to block estrogen synthesis. Fish treated with AI exhibited decreases in gonadal weight, plasma estrogen levels, and spermatogonial proliferation in the testis, together with increases in androgen levels. Additionally, we confirmed that exogenous estrogen treatments stimulated the renewal and proliferation of spermatogonia in the testis of IP males. These results indicate that estrogens play an important role in regulating spermatogenesis in this fish. Developmental Dynamics, 2011. © 2010 Wiley‐Liss, Inc.     

Quantitative (stereological) study of the effects of vasectomy on spermatogenesis in rabbits

Using stereological methods, especially the optical disector for unbiased estimation of nuclear number, our recent study demonstrated that long-term (6 or 12 months) vasectomy in the rhesus monkey had no significant effects on spermatogenesis (Peng et al. Reproduction 2002, 124, 847-856). This study aimed to determine the scenario in the rabbit using the same morphometric methodology. Three groups of normal male Japanese white rabbits (aged 4-5 months) were subjected to unilateral vasectomy; 10 days, 6 months and 12 months later both testes and epididymides were removed. Testicular and epididymal methacrylate-embedded sections were obtained for stereology. Vasectomy-induced damage to spermatogenesis was observed, primarily sloughing of spermatogenic cells with a greater reduction in the number of advanced (adluminal) cells. The damage was most severe at 10 days, occurring in all the testes on the vasectomized side and involving sloughing of even type A spermatogonia, the number of which returned to normal at 6 and 12 months. Damage was less severe at 6 and 12 months, being found in half of the testes of the vasectomy side, in which the total numbers of later germ cell types were 24.0-59.1% (spermatocytes) and 0.3-11.6% (spermatids) of control at 6 months, and 20.1-22.1% (spermatocytes) and 0.4-12.0% (spermatids) of control at 12 months. By contrast, Sertoli cell number per testis was unchanged following vasectomy in any group. Epididymis on the vasectomy side, especially at 10 days and 6 months, appeared larger than on the contralateral side, but this difference was not statistically significant, and no sperm granuloma was seen in the epididymis.