高表达FasL的Sertoli细胞在睾丸局部感染时的免疫调节作用

目的：研究高表达FasL的转基因小鼠Sertoli细胞在睾丸局部感染时的免疫调节作用。方法：以溶脲脲原体（Ureaplasma urealyticum,UU)直接注入FasL转基因的小鼠膀胱模拟上行性感染的途径，分别在1、2和3周处死小鼠，分离取得睾丸组织，观察组织的病理变化，并从小鼠睾丸组织分离获得高纯度的Sertoli细胞，然后抽提总RNA，用RT－PCR方法比较正常组与UU感染组之间FasL、IL－1α、TGF－βmRNA的表达差异。结果：与正常组相比，UU感染的小鼠，睾丸组织发生明显的病理改变，Sertoli细胞表达的调节因子在mRNA水平也发生了明显的变化：其FasLmRNA的表达在1周，3周时与正常无明显区别，2周时明显升高；IL－1α、TGF－βmRNA的表达在1－3周均为升高。结论：FasL转基因小鼠的睾丸ertoli细胞可通过改变有关细胞因子的表达格局发挥免疫调节作用。     

Sertoli细胞FasL 和TGF-β mRNA表达在感染时的免疫调节作用

目的 研究大鼠Sertoli细胞在睾丸局部感染时的免疫调节作用。方法 以解脲脲原体（UU）和致病性大肠杆菌，直接注入大鼠膀胱模拟上行性感染的途径，分别在感染后1、2、3wk处死大鼠，从大鼠睾丸组织中分离获得高纯度的Sertoli细胞。然后抽提总RNA，用RT－PCR法比较正常组与UU感染组、致病性大肠杆菌感染组之间FasL mRNA和TGF－β mRNA表达的差异性。结果 与正常组相比较在UU感染后，1－3wk FasL mRNA的表达均升高；TGF－β mRNA的表达在1、2wk时升高，3wk时下降；而致病性大肠杆菌感染后，FasL mRNA和TGF－β mRNA的表达在1－3wk时均升高。结论 大鼠Sertoli细胞在抗感染免疫中，可通过FasL mRNA和TGF－β mRNA表达的变化，调节睾丸局部的免疫功能及有助于睾丸免疫豁免的维持。     

睾丸局部感染时Sertoli细胞IL-1、IL-6、TGF-β和FasL的变化

目的：研究大鼠Sertoli细胞在抗感染中的免疫调节作用。方法：以溶脲脲原体(UU)和致病性大肠杆菌(E．coli)直接注入大鼠膀胱模拟上行性感染的途径，分别在1周、2周、3周处死大鼠，分离取得睾丸组织作组织切片观察病理变化及从大鼠睾丸组织中分离获得高纯度的Sertoli细胞，用免疫组化方法比较正常组与UU感染组、致病性大肠杆菌组之间：IL-1,IL-6,TGF-β和FasL表达的差异。结果：与正常组相比，UU和E .coli感染后，其IL-1分泌升高，IL-6分泌下降，TGF-β分泌升高，FasL表达也升高，结论：大鼠Sertoli细胞在抗感染免疫中，可通过IL-1，IL-6，TGF-β和FasL的表达来发挥免疫调节作用。     

睾丸局部感染时Sertoli细胞IL-1、IL-6 mRNA的表达

目的；研究大鼠Sertoli细胞在抗感染中的免疫调节作用。方法：以溶脲脲原体（ureaplasma urealyicum,UU)和致病性大肠杆务直接注入大肠膀胱模拟上行性感染的途径，分别在1，2和3周处死大鼠，从大鼠睾丸组织分离获得高纯度的Sertoli细胞，然后抽提总RNA，用RT－PCR方法比较正常组与UU感染组，致病性大肠杆菌组之间IL－1，IL－6 mRNA表达的差异。结果：与正常组相比，UU感染后，其IL－1 mRNA在1，2周时升高，3周时下降，IL－6在1，2周时下降，3周时升高；而致病性大肠杆菌感染后，其IL－1 mRNA在1，2周时均升高，3周下降；IL－6在1周时升高，2周时下降，3周又升高，结论：大鼠Sertoli细胞在抗感染免疫中，可能IL－1，IL－6的表达来发挥免疫调节作用。     

锌指蛋白265在大鼠睾丸Sertoli细胞中的特异性表达

目的:研究锌指蛋白265(ZNF265)在大鼠睾丸Sertoli细胞中的特异性表达.方法:分离培养SD大鼠睾丸Sertoli细胞, 以免疫荧光染色、流式细胞术(FCM)检测ZNF265在Sertoli细胞的表达, 且抽提Sertoli细胞总RNA, 用PT-PCR法检测ZNF265的表达.结果:经免疫荧光染色, 实验组Sertoli细胞核核周呈现明亮特异荧光, 对照组未见特异性荧光;FCM检测结果显示实验组Sertoli细胞ZNF265的表达较对照组明显增高;RT-PCR扩增出目的条带ZNF265.结论:大鼠睾丸Sertoli细胞特异性表达ZNF265, 且多表达于Sertoli细胞核核周胞质.     

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。     

营养性肥胖大鼠弓状核促性腺激素释放激素的表达变化以及对精子发生的影响

目的:探讨营养性肥胖大鼠弓状核神经肽Y(NPY)、瘦素受体(ob-R)及与生殖相关的促性腺激素释放激素(GnRH)表达变化以及对精子发生的影响.方法:免疫组织化学观察NPY、ob R及GnRH在肥胖模型组下丘脑弓状核的表达情况以及睾丸支持细胞雄激素结合蛋白(ABP)表达变化;流式细胞分析检测睾丸生精细胞周期的改变.并测定血清中瘦素、睾酮、卵泡刺激素(FSH)和黄体生成素(LH)的水平.结果:肥胖大鼠血清中瘦素水平较对照组明显升高,睾酮、FSH、LH水平较对照组明显降低;下丘脑弓状核NPY表达较对照组增强,ob-R及GnRH表达较对照组减弱,ABP表达较对照组减弱;肥胖大鼠S期细胞显著下降,G2/M期细胞的百分数明显增多.结论:营养性肥胖大鼠由于神经内分泌代谢失调而引起GnRH水平降低,导致下丘脑垂体睾丸轴功能失调引起睾丸间质细胞及支持细胞功能降低,致使精子发生障碍,可能导致不育.     

双氢睾酮调节原代培养大鼠Sertoli细胞GDNF的表达

目的观察双氢睾酮对原代培养大鼠Sertoli细胞GDNF及相关细胞信号通路关键蛋白的表达,探讨Sertoli细胞GDNF表达调控的机制。方法原代培养出生后20 d大鼠Sertoli细胞,给予双氢睾酮干预,免疫荧光检测GDNF在细胞中定位和表达强度,Western blot检测GDNF,ERK和CREB蛋白的表达及磷酸化情况,并用ERK和c AMP/PKA信号通路抑制剂干预。结果 GDNF表达在Sertoli细胞质中,双氢睾酮干预可增加GDNF的表达(P0.05),并使ERK和CREB磷酸化水平增高(P0.05),使用ERK和c AMP/PKA信号通路抑制剂可减低双氢睾酮诱导的GDNF表达(P0.05)。结论双氢睾酮通过激活ERK信号通路诱导Sertoli细胞GDNF的表达。     

亚慢性砷染毒对大鼠睾丸中水通道蛋白9及睾酮表达的影响

目的：研究亚慢性砷染毒对大鼠睾丸中水通道蛋白9（ AQP9）及睾酮表达的影响，为砷中毒导致的男性不 育提供理论依据。方法：雄性SD大鼠随机分为高、中、低剂量砷染毒组和对照（ 蒸馏水）组，采用经口自由饮用 方式进行染毒，连续染毒14 周。染毒结束后，取血和双侧睾丸，采用电感耦合等离子体发射光谱仪测定血浆及睾 丸中砷浓度，采用免疫组织化学法检测AQP9的表达变化， ELISA 检测血清中睾酮的浓度， H-E 染色观察睾丸组织 形态学改变。结果：与对照组比较，染砷组血浆和睾丸中有明显的砷积累。AQP9蛋白在睾丸组织中间质细胞表达 增强，中、高剂量组与对照组比较，差异有统计学意义。与对照组比较，低剂量组睾酮浓度下降，差异无统计学意义， 中、高剂量组与对照组比较，差异有统计学意义。染毒组大鼠生精上皮出现不同程度损害，睾丸间质充血、渗出等， 尤以中、高剂量组变化明显。结论：亚慢性砷中毒可以引起血浆及睾丸砷浓度增加，影响AQP9的表达，导致大鼠 睾丸组织病理学改变和睾酮分泌紊乱，对大鼠的睾丸组织和生殖功能造成损伤。     

Toll样受体对睾丸Sertoli细胞免疫调节的初步研究

目的:研究睾丸Sertoli细胞感染后,TLR发挥免疫调节功能的情况。方法:正常大鼠Sertoli细胞表达Toll样受体的情况以及用溶脲脲原体(Ureaplasma Urealyticum,UU)体外感染Sertoli细胞后,分别在12、24、36小时时间段比较感染组与对照组之间Toll样受体表达变化情况。结果:正常大鼠Sertoli细胞表达TLR2-8,低表达TLR9和10,未检出TLR1和5;与对照组相比,体外感染处理后TLR2、6表达增加。结论:Toll样受体的激活参与Sertoli细胞对炎症的免疫调节,两者具有一定的联系。     

The influence of progestin and androgen on the fine structure of the male reproductive tract of the rat. I. General effects and observations on the testis

The combination of a progestin and androgen has received attention as a possible male contraceptive. The progestin is thought to reduce gonadotropin release and suppress spermatogenesis, while the sex accessory organs and male characteristics are maintained by the simultaneous administration of testosterone. In the present study, the histology and ultrastructure of parts of the male reproductive tract of rats treated with medroxyprogesterone (Provera, Upjohn) (1 mg/100 g body weight/day) alone and combined with testosterone (15, 30, or 100 μg/100 g/day) were studied following treatment for up to 16 weeks. The testes and epididymides of rats administered Provera alone or Provera and testosterone weighed less than those of control rats. The weights of the accessory glands of rats treated with Provera were greatly reduced; it was possible to maintain them at approximately control levels by simultaneously administering sufficient testosterone (100 μg/100 g body weight/day). The fertility of some of the animals was tested by caging them with female rats, and none of the treated rats tested in this way was fertile. Similar microscopic alterations were present in the testes of animals administered Provera alone or Provera and different levels of testosterone. Spermato-gonia, spermatocytes, and early spermatids were abundant in treated rats and did not show ultrastructural changes. However, many degenerating or necrotic spermatids of the cap phase (approximately stages 6–7 ) and later were present. Late spermatids of the acrosome and maturation phases were rare. Some necrotic spermatids were surrounded by Sertoli cells, and parts of spermatids lay within lysosome-like structures in the cytoplasm of Sertoli cells. Many large lipid droplets were also present in Sertoli cells of treated rats. Leydig cells were smaller in treated animals than in control rats. The results suggest that germ cells can develop up to cap phase spermatids but then undergo degeneration. These alterations in spermatogenesis may be responsible in large part for the antifertility effect of the progestin and androgen combination. Some rats were permitted to recover following the end of treatment. The microscopic appearance of the testis returned to normal within three to six weeks, although epididymal alterations persisted in some animals six weeks after the end of treatment. By 9 to 12 weeks after the end of treatment the reproductive organs had a normal microscopic appearance in all the rats studied.     

Cell-cell interactions in the control of spermatogenesis as studied using leydig cell destruction and testosterone replacement

This review centers around studies which have used ethane dimethane sulphonate (EDS) selectively to destroy all of the Leydig cells in the adult rat testis. With additional manipulations such as testosterone replacement and/or experimental induction of severe seminiferous tubule damage in EDS-injected rats, the following questions have been addressed: (1) What are the roles and relative importance of testosterone and other non-androgenic Leydig cell products in normal spermatogenesis and testicular function in general? (2) What are the factors controlling Leydig cell proliferation and maturation? (3) Is it the Leydig cells or the seminiferous tubules (or both) which control the testicular vasculature? The findings emphasize that in the normal adult rat testis there is a complex interaction between the Leydig cells, the Sertoli (and/or peritubular) cells, the germ cells, and the vasculature, and that testosterone, but not other Leydig cell products, plays a central role in many of these interactions. The Leydig cells drive spermatogenesis via the secretion of testosterone which acts on the Sertoli and/or peritubular cells to create an environment which enables normal progression of germ cells through stage VII of the spermatogenic cycle. In addition, testosterone is involved in the control of the vasculature, and hence the formation of testicular interstitial fluid, presumably again via effects on the Sertoli and/or peritubular cells. When Leydig cells regenerate and mature after their destruction by EDS, it can be shown that both the rate and the location of regenerating Leydig cells is determined by an interplay between endocrine (LH and perhaps FSH) and paracrine factors; the latter emanate from the seminiferous tubules and are determined by the germ cell complement. Taken together with other data on the paracrine control of Leydig cell testosterone secretion by the seminiferous tubules, these findings demonstrate that the functions of all of the cell types in the testis are interwoven in a highly organized manner. This has considerable implications with regard to the concentration of research effort on in vitro studies of the testis, and is discussed together with the need for a multidisciplinary approach if the complex control of spermatogenesis is ever to be properly understood.     

Antireproductive effect of the calcium channel blocker amlodipine in male rats.

The purpose of the present study was to investigate whether treatment of male rats with the calcium antagonist amlodipine, used in the treatment of hypertension and angina, interferes with the reproductive function of male rats. Amlodipine treatment (0.04 mg amlodipine besylate/rat/day for 30 days) decreased plasma follicle-stimulating hormone and testosterone but not luteinizing hormone or prolactin concentrations (measured by double-antibody radioimmuno-assay). A significant reduction (23%) was observed in sperm density (sperm suspension collected from the cauda epididymidis) as well as in the amount of mature spermatids (14%) and Sertoli cells (9%) counted in seminiferous tubule cross-sections (400 x magnification). The results reveal the deleterious effects of subacute amlodipine treatment on the reproductive function of male rats.     

维药伊木萨克片对大鼠阴茎勃起功能的影响

目的:探讨维药伊木萨克片对雄性大鼠阴茎勃起功能的影响及其机制.方法:性功能正常SD雄性大鼠用伊木萨克连续干预6周后,进行阿普吗啡(APO)阴茎勃起实验和交配实验,并检测阴茎海绵体窦压(ICP);用放射免疫法检测外周血清中睾酮(T)、促黄体生成素(LH)和促卵泡刺激素(FSH)含量,镜检睾丸的组织形态学改变;用免疫组织化学技术检测两组大鼠阴茎组织中eNOS、nNOS表达.结果:(1)与正常对照组相比较,伊木萨克组APO勃起所需时间显著缩短,插入次数显著增加,且基础ICP值与电刺激诱导ICP值均显著升高;(2)T水平在伊术萨克组显著高于正常对照组,而LH与FSH在两组问无显著性差异;(3)eNOS表达在伊木萨克组显著高于正常对照组,nNOS在两组间无昆著性差异.结论:维药伊木萨克片能够显著提高正常大鼠的阴茎勃起功能,其机制可能和提高雄激素水平与eNOS表达有关.     

丝胶对2型糖尿病大鼠睾丸生长激素/胰岛素样生长因子-1轴的作用

目的 探讨丝胶对2型糖尿病大鼠睾丸生长激素（GH）/胰岛素样生长因子-1（IGF-1）轴的作用。 方法 40只雄性SD大鼠随机分为正常对照组、糖尿病模型组、丝胶治疗组和阳性对照组,每组均为10只。链脲佐菌素连续腹腔注射制作2型糖尿病大鼠模型,1周后以血糖≥16.7mmol/L作为成模标准。待模型成功建立后,模型组大鼠不做任何处理,丝胶治疗组大鼠给予丝胶灌胃［2.4g/(kg•d)］,阳性对照组大鼠给予二甲双胍［55.33mg/(kg•d)］灌胃,均为35d。采用酶联免疫吸附测定（ELISA）方法检测大鼠血清睾酮、GH和IGF-1水平;分别采用免疫组织化学染色、免疫印迹法和RT-PCR法检测睾丸GH、GH受体（GHR）和IGF-1的表达。 结果 丝胶可明显降低糖尿病大鼠血GH水平、下调睾丸GH的表达,升高血IGF-1和睾酮水平,上调睾丸IGF-1和GHR的表达（P＜0.05,P＜0.01）。并且丝胶治疗组各项指标与阳性对照组比较无明显差别（P＞0.05）。 结论 丝胶可通过调节糖尿病时GH/IGF-1轴紊乱改善生精功能,发挥对糖尿病生殖功能损害的保护作用,其作用与二甲双胍相当。     

Expression of CD1d Protein in Human Testis Showing Normal and Abnormal Spermatogenesis

CD1d is a member of CD1 family of transmembrane glycoproteins, which represent antigen-presenting molecules. Immunofluorescent staining methods were utilized to examine expression pattern of CD1d in human testicular specimens. In testis showing normal spermatogenesis, a strong CD1d cytoplasmic expression was seen the Sertoli cells, spermatogonia, and Leydig cells. A moderate expression was observed in the spermatocytes. In testes showing maturation arrest, CD1d expression was strong in the Sertoli cells and weak in spermatogonia and spermatocytes compared to testis with normal spermatogenesis. In Sertoli cell only syndrome, CD1d expression was strong in the Sertoli and Leydig cells. This preliminary study displayed testicular infertility-related changes in CD1d expression. The ultrastructural changes associated with with normal and abnormal spermatogenesis are open for further investigations.     

Expression of CD1d protein in human testis showing normal and abnormal spermatogenesis

CD1d is a member of CD1 family of transmembrane glycoproteins, which represent antigen-presenting molecules. Immunofluorescent staining methods were utilized to examine expression pattern of CD1d in human testicular specimens. In testis showing normal spermatogenesis, a strong CD1d cytoplasmic expression was seen the Sertoli cells, spermatogonia, and Leydig cells. A moderate expression was observed in the spermatocytes. In testes showing maturation arrest, CD1d expression was strong in the Sertoli cells and weak in spermatogonia and spermatocytes compared to testis with normal spermatogenesis. In Sertoli cell only syndrome, CD1d expression was strong in the Sertoli and Leydig cells. This preliminary study displayed testicular infertility-related changes in CD1d expression. The ultrastructural changes associated with with normal and abnormal spermatogenesis are open for further investigations.     

高血压合并糖尿病大鼠心肌毛细血管内皮细胞超微结构及eNOS的变化

目的:探讨高血压合并糖尿病(DM)对心肌毛细血管内皮细胞超微结构及内皮型一氧化氮合酶(eNOS)表达的影响。方法:自发性高血压大鼠(SHR)及SD大鼠腹腔注射链脲佐菌素(STZ)结合高能量饲料诱导DM模型。分为4组:正常SD大鼠组(SD组)、SHR组、单纯DM组(DM组)与自发性高血压合并DM大鼠组(SHDM组)。电镜观察各组心肌毛细血管内皮细胞超微结构,免疫组化法测定各组eNOS在毛细血管内皮细胞的表达。结果:SHR、DM和SHDM组心肌毛细血管超微结构明显改变,包括内皮细胞水肿,细胞膜向管腔内呈指状突起,管腔狭窄、不规则等,这些现象在DM组与SHDM组更明显。与SD组比较,SHR组基底膜稍有增厚,但无显著性差异(31.31±4.19nmvs28.64±3.62nm,P>0.05),而DM组(46.58±5.32nm)和SHDM(51.50±4.62nm)组基底膜显著增厚(与SD及SHR组比较,均P<0.01)。SHDM组心肌组织eNOS表达显著低于SHR组及DM组。结论:高血压与DM共存时对心肌毛细血管内皮细胞超微结构及功能有协同损害作用。     

成人睾丸支持细胞的分离培养及其免疫豁免机制

目的：建立成人睾丸支持细胞的体外培养方法并探讨其免疫豁免机制。方法：依次用胰蛋白酶、胶原酶、透明质酸酶及脱氧核糖核酸酶消化制备成人睾丸支持细胞，以SABC染色法检测支持细胞上Fas-L和TGF-β1的表达。将其与成人脾细胞共同培养，用MTT比色法检测其对脾细胞增殖的抑制作用。结果：成人睾丸支持细胞占培养细胞总数的80％以上，活性可达90％。睾丸支持细胞上可表达Fas-L和TGF-β1，体外可抑制共培养的脾细胞增殖。结论：建立了培养成人睾丸支持细胞的方法，其免疫豁免机制可能与Fas-L和TGF-β1的表达有关。     

环孢素A转换为雷帕霉素长期作用对移植肾大鼠睾丸功能和形态学的影响

比较环孢素A(CsA)转换为雷帕霉素(Rapa)后与CsA长期作用对移植肾大鼠睾丸功能和组织形态学的影响。方法 采用标准的肾移植模型方法进行原位左肾移植,即将Fisher大鼠的供肾移植给Lewis雄性大鼠,36只大鼠分为CsA转换为Rapa组(R组,n=10)、CsA持续使用组(A组,n=10)、CsA撤离组(B组,n...