褪黑激素对小鼠睾丸生精细胞凋亡和睾丸间质细胞nNOS表达的影响

目的：观察外源性褪黑激素(MT)对睾丸生精细胞凋亡和nNOS表达的影响,探讨MT诱导生精细胞凋亡的可能机制。方法：20d和30d小鼠,应用化学发光法检测血清睾酮浓度,TUNEL测定生精细胞凋亡,免疫组化,SABC法观察nNOS在睾丸中的表达。结果：20d和30d实验组血清中睾酮水平明显下降,TUNEL阳性生精细胞数显著增多,TUNEL阳性生精细胞数与睾酮浓度呈负相关;20d实验小鼠中睾丸间质nNOS阳性细胞数与对照组相比明显减少,且与睾酮浓度呈正相关,而与TUNEL阳性生精细胞数呈负相关。结论：MT具有促进生精细胞凋亡的作用,与睾酮分泌受抑制有关;青春期前MT引起生精细胞凋亡可能还与nNOS表达有关。     

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。     

睾丸波形蛋白表达与生精细胞凋亡的关系

睾丸的生精上皮具有高度增殖能力,生精过程中细胞增殖与凋亡的动态平衡,对于维持精子的生成的数量和质量具有重要的作用。外来化学毒物可以直接引起睾丸损伤,导致生精细胞过量凋亡,从而导致精子生成减少。生精细胞凋亡时细胞骨架蛋白改变的研究成为一个热点,睾丸波形蛋白表达发生了异常改变,但睾丸波形蛋白表达与生精细胞凋亡的相关性还需进一步研究。     

人类睾丸生精细胞凋亡相关基因TSARG3的克隆

目的：克隆人类睾丸生精细胞凋亡相关基因TSARG3。方法：从已获得的小鼠稳睾和正常睾丸对照中表达量有明显差异的表达序列标签片段（BE644537）入手，构建人同源表达序列标签重叠群，应用基因特异性引物和载体特异性引物，在睾丸cDNA文库的DN或进行巢式PCR扩增、测序，对测序结果进行生物信息学分析。结果：从睾丸cDNA文库中分离出人类睾丸凋亡相关基因的5’末端而获得全长cDNA,命名为TSARG3，GenBank登录号为AF419291（保密期为1年），同时应用相同方法克隆了该基因在小鼠中的同源基因，GenBank登录号为AF419292。结论：获得人类睾丸生精细胞凋亡相关基因TSARG3，该基因可能与人类睾丸生精细胞凋亡有关。     

小鼠肾脏发育中凋亡细胞的超微结构变化

1　材料和方法1 1　动物取昆明小鼠 ,雌雄比例 3∶1同窝饲养 ,以雌鼠阴道精栓出现为第 1日计算胎鼠胚龄。部分孕鼠分笼饲养 2 0日后 ,每天早八时观察分娩情况 ,新生仔鼠按生后计算日龄。选取 1 4、1 8日胎鼠各 4只 ,生后 1、7、1 4日龄仔鼠各 4只为观察对象。1 2　光镜和电镜标本制作　在冷操作条件下切开胎鼠和仔鼠腹部 ,取左肾于 2 5 %戊二醛内固定。胎鼠做全肾树脂包埋 ,仔鼠肾脏经肾门做矢状面切开 ,皮质和髓质分别进行树脂包埋。ＬＫＢ Ｖ型超薄切片机半薄连续切片 ,甲苯胺蓝染色 ,按Ｋｏｓｅｋｉ判断光镜下凋亡细胞后制超薄切片 ,…     

睾丸间质细胞及淋巴管的超微结构

目的　探明睾丸间质细胞及淋巴管的超微结构。方法　灌流固定后半薄超薄切片光镜和电镜下观察。结果　睾丸间质细胞有二种亚群 ,各有其特点。淋巴管存在于纵隔内 ,小叶内未见典型淋巴管。结论　睾丸间质细胞分明细胞和暗细胞二种 ,纵隔内有丰富的淋巴管及毛细淋巴管 ,而小叶内则存在广泛的淋巴间隙     

饮酒对小鼠睾丸生精小管一氧化氮合酶、增殖细胞核抗原表达及细胞凋亡影响的研究

目的　探讨酒精对小鼠睾丸的组织结构、内皮型一氧化氮合酶 (eNOS)、增殖细胞核抗原 (PCNA)及细胞凋亡的影响。　方法　用 5 %、10 %及 15 % 3种不同浓度的酒精作用于 2 2d龄小鼠 ,取睾丸做石蜡切片、HE染色 ;用免疫组织化学方法检测睾丸eNOS、细胞增殖的变化 ;TUNEL法检测细胞凋亡的变化 ,并进行统计学分析。　结果　随着酒精浓度的增大 ,睾丸组织结构发生明显改变 ,生精小管的直径逐渐减小 ,eNOS阳性细胞面积密度逐渐增大 ,单位面积内PCNA阳性细胞和凋亡细胞数目增加 ,高浓度酒精组与其他组差异极显著 (P <0 0 1)。　结论　酒精可使生精小管的直径变小 ,eNOS及PCNA表达增强 ,凋亡细胞增加并随酒精浓度的增大而变化加重。这可能是过量饮酒导致生精细胞减少 ,生殖能力降低的重要因素。     

过量饮酒对凋亡相关基因bcl-2、bax在生精细胞表达的影响

目的　研究长期过量饮酒对凋亡相关基因bcl - 2、bax在睾丸及生精细胞中表达的影响。方法　利用人类饮用白酒制备大鼠的酒精毒性模型 ,采用免疫组织化学技术 ,检测酒精对Bcl- 2、Bax蛋白在睾丸及生精细胞表达的影响。结果　高剂量的酒精作用于大鼠 2个生精周期 ,每个生精小管中Bcl- 2蛋白表达的阳性细胞数目在高剂量组为 96± 33.74 ,低剂量组为 15 6 .6± 2 8.5 2 ,均显著低于正常对照组 (2 2 2 .6± 5 6 .86 ) (P <0 .0 1)。每个细胞中Bcl- 2蛋白表达强度 (OD值 )在高剂量组 (0 .14 2± 0 .0 35 )、低剂量组 (0 .15 1± 0 .0 2 6 ) ,都明显低于对照组 (0 .196± 0 .0 32 ) (P <0 .0 1) ;Bax蛋白表达的阳性细胞数在高剂量组为 (4 12 .4± 96 .75 ) ,低剂量组为 (337.5± 94 .39) ,均明显高于对照组 (2 14± 82 .5 7) (P <0 .0 1) ,Bax蛋白的表达强度 (OD值 )在高剂量组 (0 .113± 0 .0 36 )和低剂量组 (0 .0 82± 0 .0 17) ,均明显高于对照组 (0 .0 5± 0 .0 2 4 ) (P <0 .0 1)。结论　长期过量饮酒使睾丸及生精细胞中bcl- 2的表达降低 ,bax的表达增强。     

Effect of extract of Hibiscus on the ultrastructure of the testis in adult mice

Hibiscus sabdariffa extract is a popular beverage in many tropical and sub-tropical countries. Although, Hibiscus tea is known for its medicinal effects for thousands of years, scientific evidence of its systemic safety is very limited. The current study aimed to assess the potential adverse effects of H. sabdariffa extract on sperm morphology and testicular ultrastructure of albino mice. Thirty adult male albino mice were divided into three equal groups and were given: (a) distilled water, (b) cold Hibiscus aqueous extract, and (c) boiled Hibiscus aqueous extract. Hibiscus extract was administered orally daily for 4 weeks in a dose of 200 mg/kg body weight/mouse. Twenty-four hours after the last treatment, mice were decapitated and the testes and epididymides were excised and processed for transmission electron microscopy to assess ultrastructural and sperm abnormalities. The results clearly demonstrate that aqueous extracts from dried calyx of H. sabdariffa, either cold or boiled, alter normal sperm morphology and testicular ultrastructure and adversely influence the male reproductive fertility in albino mice. The current data suggest that Hibiscus extract should be consumed with caution, and reasonable estimates of the human risk associated with its consumption should be provided.     

衰老大鼠睾丸生精细胞增殖凋亡及相关因素的变化

目的:观察衰老大鼠睾丸生精细胞增殖、凋亡及相关因素的变化。方法:D-半乳糖连续腹腔注射制作亚急性衰老的大鼠模型。SP免疫组织化学法观察睾丸生精细胞衰老过程中增殖细胞核抗原(PCNA)和端粒酶的表达,TUNEL法观察生精细胞的凋亡,原位杂交、Western印迹分析观察生精细胞p53基因的表达。结果:与正常大鼠相比,衰老大鼠睾丸PCNA阳性细胞率显著下降,端粒酶阳性细胞数明显减少,凋亡管数和凋亡阳性细胞率显著升高,p53 mRNA阳性细胞率和p53蛋白含量均有所升高。结论:衰老大鼠睾丸生精细胞增殖能力下降、凋亡增多,睾丸机能减退。     

LPS-induced transient testicular dysfunction accompanied by apoptosis of testicular germ cells in mice

The aim of this study was to clarify effects of inflammation on spermatogenesis in LPS-administered mice. ICR mice were treated by intraperitoneal injection for 7 days with either physiological saline (control) or 0.1 mg lipopolysaccharide (LPS)/kg body weight/day. Control mice were killed at 24 h after the last injection and the LPS-treated group after 24 h or 1, 3, or 5 weeks. Sperm concentration and motility in the cauda epididymis were examined as well as immunohistochemical localization of Fas and FasL and germ cell apoptosis. Sperm concentration and motility markedly fluctuated in LPS-treated mice. Increase of apoptotic cells was common in all post-LPS treatment groups, with a peak at 24 h after LPS injection. In contrast to the lack of Fas immunoreactivity in control testes, LPS-treated groups demonstrated Fas in many germ cells, especially in spermatocytes and spermatids. Immunoreactivity for FasL, on the other hand, was positive for some Sertoli cells, Leydig cells, and germ cells in both control and LPS-treated groups at all time points. The results suggest that the Fas/FasL system mediates apoptosis of germ cells in LPS-treated mice testes. LPS-administered mice thus provide a good experimental model for the study of transient disruption of spermatogenesis.     

葛根素减轻乙醇导致大鼠生精细胞的凋亡

目的 观察乙醇导致大鼠生精细胞的凋亡及葛根素的干预。方法 将大鼠30 只,随机均分为对照组、乙醇组及葛根素干预组。于实验第40天免疫组织化学法（SABC）检测左侧睾丸各组Bcl-2、Bax 蛋白在生精细胞的表达; RT-PCR检测右侧睾丸各组Bcl-2及Bax mRNA的表达;TUNEL 法检测生精细胞的凋亡。结果 醇组平均每个生精小管断面中Bcl-2 蛋白阳性细胞数和A值低于对照组(P<0.01),而平均每个生精小管断面中Bax 蛋白的阳性细胞数和A值高于对照组(P<0.01);乙醇组Bax mRNA表达较葛根素干预组及对照组强（P<0.05）,而Bcl-2 mRNA表达较葛根素干预组及对照组弱（P<0.05）;乙醇组每个生精小管横切面中的凋亡细胞数目高于对照组(P< 0.01),葛根素干预组显著缓解上述变化。结论 根素对乙醇导致的大鼠生精细胞凋亡有干预作用。     

茶多酚对精索静脉曲张大鼠生精细胞凋亡的影响及机制研究

目的探究茶多酚(TP)对精索静脉曲张(Vc)SD大鼠生精细胞凋亡及细胞色素c(Cyt-c)、波形蛋白表达的影响及机制。方法将36只SD大鼠分为对照组、Vc组和Vc+TP组,每组12只。Vc组和Vc+TP组通过左肾静脉结扎构建精索静脉曲张模型,Vc+TP组大鼠给予茶多酚(40 mg/kg)灌胃,对照组和Vc组以等量生理盐水灌胃,连续4周。干预4周后取出大鼠睾丸组织,通过HE染色和TUNEL染色观察睾丸组织损伤和生精细胞凋亡情况,并检测茶多酚对氧化应激指标的影响。通过RT-qPCR和Western blot检测Cyt-c和波形蛋白表达。结果HE染色结果显示,Vc组出现广泛的上皮损伤,Vc+TP组损伤情况明显轻于Vc组。Vc组凋亡指数高于对照组及Vc+TP组(P<0.05)。Vc组丙二醛(MDA)水平高于对照组及Vc+TP组(P<0.05),而超氧化物歧化酶(SOD)水平低于对照组及Vc+TP组(P<0.05);Vc组波形蛋白mRNA和蛋白表达水平均低于对照组及Vc+TP组(P<0.05),而Cyt-c mRNA和蛋白表达水平均高于对照组及Vc+TP组(P<0.05)。结论茶多酚可通过抑制氧化应激反应保护线粒体功能并促进波形蛋白的表达,从而抑制生精细胞凋亡,减轻睾丸损伤,达到缓解精索静脉曲张的目的。     

Effects of mouse testicular extract on immunocompetent cells

We investigated mouse testicular extract (TE) to clarify its biological functions in reproductive immunity. TE, at concentrations of 50-300 micrograms/ml, enhanced macrophage activities of spreading, glucose consumption, and cytostasis against a susceptible tumor cell line. On the other hand, TE inhibited concanavalin A (Con A)-induced T-cell blastogenesis in the dose range of 10-600 micrograms/ml. To elucidate the origin of TE, W/Wv mice, which genetically lack germ cells, were used. TE obtained from W/Wv mice enhanced the spreadability of macrophages and inhibited Con A-induced blastogenesis of T cells. The enhancement of macrophage spreading was only achieved by the interstitial fluid (IF), while the suppression of Con A-induced T-cell responses was detected in seminiferous tubule fluid (STF) as well as in IF. TE did not affect listerial antigen-specific responses of lymphocytes in vitro. These results suggest that TE has the capacity to regulate the biological responses associated with reproduction.     

N-硝基-L-精氨酸甲酯对大鼠隐睾生精细胞凋亡的影响

目的：研究N-硝基-L-精氨酸甲酯(L-NAME)对实验性大鼠隐睾生精细胞凋亡的影响及其作用机制。方法：手术建立大鼠单侧隐睾模型,术后分别注射L-NAME及生理盐水,7d后采用流式细胞术检测2组大鼠隐睾生精细胞凋亡,免疫组化法检测隐睾内Bcl-2和Bax基因表达变化。结果：实验组隐睾重量较对侧正常睾丸重量减轻的程度低于对照组,生精细胞凋亡百分比及Bax表达较对照组低,而Bcl-2表达较对照组高。结论：L-NAME可通过调控Bcl-2和Bax的表达抑制隐睾导致的生精细胞凋亡。     

高血黏度对小鼠脑细胞线粒体含量及超微结构的影响

目的：观察高血黏度对脑细胞线粒体含量及超微结构的影响。方法：采用高分子右旋糖酐法建立小鼠高血黏度动物模型,以Mito Tracker Green FM对线粒体进行标记,用激光扫描共聚焦显微镜测定脑细胞线粒体含量的变化;透射电镜观察小鼠大脑顶叶皮质及海马CA1区神经元线粒体超微结构的变化。结果：高血黏度小鼠脑细胞线粒体含量非常明显地减少;线粒体超微结构发生明显变化,且海马CA1区较皮质部变化明显。结论：高血黏度可导致脑细胞线粒体含量的减少及超微结构的变化。