Ⅱ型糖尿病雄性大鼠血清、生殖器官生物化学及组织学变化的实验研究

目的 :观察和探讨 型糖尿病雄性大鼠血清、睾丸、附睾和阴茎中 NO、NOS、SOD和 MDA含量的变化和相互关系 ,及其对雄性大鼠生殖器官的影响。方法 :4～ 6月龄 Wistar雄性大鼠 30只 ,随机分为 型糖尿病模型组 2 0只和对照组 1 0只。饲以高糖 -高脂饲料后 ,取血清及组织 ,光镜下观察各组织的病理改变 ,并测定各指标的变化。结果 : 型糖尿病大鼠模型制备成功 ;糖尿病大鼠与正常比较 ,显示睾丸曲细精管数量减少 ,管壁呈纤维样变 ;睾丸间质小血管内皮细胞增生 ,管壁纤维组织增生显著 ;附睾管腔内精子数量减少或缺如 ;阴茎海绵体平滑肌数量减少 ,纤维结缔组织增生。糖尿病大鼠血清及生殖器官中 NO含量和 NOS、SOD活性均显著低于对照组 ,MDA浓度明显高于对照组。相关分析表明 ,睾丸中 SOD活性与 MDA含量呈负相关 ,NO含量与 NOS活性呈正相关。结论 :II型糖尿病时睾丸、附睾及阴茎组织均受到不同程度的损害 ;糖尿病时自由基产生增加及清除障碍导致血清及生殖器官中 MDA、SOD与 NO、NOS发生变化 ,且这些变化可导致睾丸、附睾和阴茎功能紊乱 ,进而造成男性不育     

腺嘌呤诱导不育大鼠精子中精蛋白mRNA的研究

目的 :研究正常生育大鼠和腺嘌呤诱导的不育大鼠附睾精子中精蛋白及其 m RNA的含量以及它们的相关性。方法 :大鼠喂服腺嘌呤制备不育动物模型 ,收集对照组和不育组大鼠附睾精子 ,提取总碱性核蛋白 ( TNBP) ,电泳分析 ;提取总 RNA,用 RT- PCR分析精蛋白 m RNA相对含量。结果 :不育组大鼠附睾精子中精蛋白含量明显低于正常组 ( P<0 .0 1 ) ,对精蛋白 m RNA的分析也得到了相似结果 ,正常组和不育组相比较 ,有非常显著差异 ( P<0 .0 0 1 )。结论 :精子中精蛋白 m RNA的相对含量能反映出精子中精蛋白的相对含量 ,为男性不育症的基因诊断提供了新思路     

雷公藤多甙对大鼠生精细胞及其酶活性的影响

本文连续观察了雄性大鼠服用雷公藤多甙(GTW)30至80d 后睾丸、附睾细胞形态学改变及睾丸 ACP、ALP、3β-HSD、睾丸和附睾生精细胞 LDH-C_4活性的变化;同时注意了药物抗生育作用的可逆性。结果表明:附睾精子先于睾丸生精细胞发生质和量的变化;支持细胞 ACP 活性有增强趋势;睾丸 ALP,LDH-C_4酶活性相对减弱,为生精细胞损伤的结果;用药80d 时,3β-HSD 活性则明显减弱。结果还提示 GTW 引起的不育似有恢复的可能。     

少量人类精子以空卵透明带为冻贮载体的冷冻保存

目的:探讨空卵透明带作贮存载体冷冻保存少量人类精子。方法:将人或金黄地鼠卵内的细胞成份全部除去,制备成空卵透明带,分别显微注入人睾丸精子、附睾精子和射出精子后冷冻保存,并与正常供精者射出精子作对照。结果:解冻后卵透明带在溶液中容易识别寻找,卵透明带内的精子易于观察。附睾精子组(n=11)与供精者射出精子组(n=7)的冷冻精子活动率和存活率无显著性差异(P>0.05),但睾丸精子组(n=7)这两项参数值均显著低于附睾精子和射出精子组(P<0.01)。含6％、7.5％和9％不同甘油浓度的冷冻保护剂,对冷冻精子活动率无显著性影响(P>0.05);冻贮于人和金黄地鼠空卵透明带内的精子,两者的冷冻精子活动率也无显著性差异(P>0.05〕。结论:空卵透明带是冻贮少量人类精子的合适载体。     

年龄因素对大鼠睾丸中脑红蛋白表达的影响

目的:探讨脑红蛋白(neuroglobin,NGB)在不同年龄段大鼠睾丸中的分布特征及表达差异。方法:随机选取3月、9月、15月龄SD雄性大鼠,每个年龄段各8只,处死后立即在无菌情况下取睾丸组织,免疫组织化学SP法观测NGB在大鼠睾丸中的定位,巢式PCR和Western blotting分析NGB mRNA和蛋白的表达差异。结果:免疫组织化学显示NGB在大鼠睾丸的间质细胞、精原细胞、各级精母细胞、支持细胞和精子细胞的胞质和胞核都有表达;巢式PCR检测表明9月龄组大鼠睾丸组织中NGB mRNA表达量最高,15月龄组表达次之,3月龄组表达量最低;而NGB蛋白表达与其mRNA相一致。结论:NGB在各年龄段大鼠睾丸组织中广泛表达,且年龄因素影响大鼠睾丸中脑红蛋白的表达。     

L-肉碱在奥硝唑致雄性大鼠生殖系统氧化损伤中的作用

目的:探讨L-肉碱(LC)在奥硝唑(ORN)所致雄性大鼠生殖系统氧化损伤中的保护作用。方法:40只雄性SD大鼠随机均分为5组,分别连续灌胃20d。A组(对照组):0.5%羧甲基纤维素钠(溶剂);B组:400mg/kgORN;C组:800mg/kgORN;D组:ORN(400mg/kg)+LC(100mg/kg);E组:ORN(800mg/kg)+LC(100mg/kg)。末次给药24h后,麻醉处死所有大鼠,分别检测各组睾丸、附睾组织匀浆中丙二醛(MDA)、超氧化物歧化酶(SOD)水平及总抗氧化能力(TAC)。结果:B组、C组及E组大鼠附睾MDA水平明显高于A组(P均<0.01),睾丸中MDA水平则各组间没有差异;B组、C组及E组睾丸、附睾SOD水平和TAC则分别显著低于A组(P均<0.05),而D组与A组相比无显著性差异;与C组相比,E组大鼠附睾MDA水平显著降低(P<0.01),而TAC则显著升高(P<0.05);虽然E组大鼠附睾、睾丸SOD水平及睾丸中的TAC与C组比无统计学差异,但有升高的趋势。结论:LC可保护大鼠生殖系统对抗ORN所致的氧化损伤,对大鼠生殖功能具有一定的保护作用。     

亚慢性口服氯化镉对成年大鼠睾丸生精功能的影响

目的:探讨亚慢性口服氯化镉(CdCl2)对成年SD大鼠睾丸生精功能的影响。方法:将SD大鼠分成8组,每组6只,分别喂含Cd2+0mg/kg(A组),8mg/kg(B组),40mg/kg(C组),200mg/kg(D组)的全价鼠粮1个月,及含Cd2+0mg/kg(E组),8mg/kg(F组),40mg/kg(G组),200mg/kg(H组)的全价鼠粮2个月。采用光镜和透射电镜观察睾丸生精上皮的组织学和细胞学变化,以精子常规参数来评价精子品质的改变。结果:光镜观察见D组和H组曲细精管初级精母细胞层与精原细胞层之间发生分离,局限性生精上皮脱落;H组生精上皮细胞层次减少。透射电镜观察到D组支持细胞胞质电子密度减小,残余小体增多,精原细胞核固缩,胞质内出现空泡。H组还出现曲细精管界膜内陷,支持细胞次级溶酶体、脂滴和残余小体增多,精子尾部中段外周致密纤维及外周微管部分缺失。D组体重增长值、睾丸每日精子生成量、附睾尾精子量与A组相比明显减少(P<0.05)。G和H组睾丸每日精子生成量、附睾尾精子量明显少于E组(P<0.01)。睾丸每日精子生成量、附睾尾精子量与染Cd2+剂量呈明显负相关关系(P<0.001)。H组与E组相比,体重增长下降,睾丸水肿,精子存活率、活动率和精子正常形态率下降,精子畸形指数上升(P<0.05)。结论:亚慢性口服CdCl2可造成成年大鼠睾丸生精上皮损伤,生精功能下降,损伤具有?     

β-趋化性细胞因子RANTES在人睾丸和附睾中的表达

目的:探讨受激活调节正常T细胞表达和分泌因子(RANTES)在成人睾丸及附睾组织中的表达及分布。方法:提取人睾丸和附睾总RNA,进行RT-PCR分析RANTES的表达;同时取睾丸和附睾组织固定,石蜡包埋,用免疫组化法检测RANTES在人睾丸和附睾组织中的定位情况。结果:在睾丸中未见RANTES的表达,而从附睾组织中获得了RANTES的cDNA片段,且RANTES蛋白定位于附睾输出小管的高柱状细胞和低柱状细胞靠近腔面的胞质内,附睾头部远端及体、尾部均未检测到RANTES蛋白。结论:RANTES在人的附睾中有表达,且其分布表现出明显的附睾区域特异性,提示RANTES基因可能参与精子成熟过程的调控。     

雷公藤单体(TW_(19))雄性抗生育活性的研究

雷公藤单体(TW_(19))系从雷公藤中分离、提纯出的一个二萜类化合物,本实验观察其对雄性大鼠抗生育作用及可能机理.TW_(19)(400μg/kg·d)给药三周后无抗生育作用,五周后生育力丧失,附睾尾部精子活力和密度明显降低,畸形率升高,附睾重量减轻,但对附睾上皮的形态变化无明显影响,管腔内可见畸形精子及脱落生精细胞.附睾尾部液中肉毒碱含量,给药组较对照组显著降低.TW_(19)对附睾头部α-糖苷酶和酸性磷酸酶含量没有明显影响;对生精上皮影响轻微,各期曲细精管基本正常,少数表现出精子细胞轻度损伤;TW_(19)可使睾丸重量减轻,并可显著降低睾丸透明质酸酶含量,但对睾丸乳酸脱氢酶-C_4没有明显抑制作用;对前列腺、储精囊等附属性腺器官重量没有影响.结论:TW_(19)对雄性大鼠具有确定的抗生育效果,作用环节可能在精子细胞及附睾.     

不育症患者精子X,Y及18染色体的荧光原位杂交分析

目的:分析不育症患者精子X,Y,18染色体的荧光原位杂交情况。方法:在男性不育症组中,2例无精子症患者从附睾抽吸获取精子、3例从睾丸获取精子、2例严重少精症患者从射出精液中找到精子。选择5例正常射出精液者作为对照组。应用荧光原位杂交法(FISH)检查精子X,Y以及18号染色体,比较两组精子染色体非整倍体的发生率。结果:不育症组睾丸精子、附睾精子的非整倍体率无差别,但不育症组精子与正常男性组精子比较,非整倍体总发生率、性染色体二体性率及缺对染色体率明显增高(2.8％vs0.58％,0.81％vs 0.19％,2.1％vs0.37％),P<0.001。结论:无精子症与严重少精子症患者的精子比正常精子具有更高的染色体非整倍体率,需要进行大样本的研究,为不育症患者的治疗和遗传咨询提供有效的证据。     

Effect of protopanaxatriol saponin on spermatogenic stem cell survival in busulfan-treated male mice

Background and aims:  Panax ginseng C.A. Meyer is a medicinal herb widely used in Asian countries. Many of its pharmacological actions are attributed to ginsenosides (saponin). However, the pharmacological effects or functions of ginsenosides on mammalian spermatogenesis are unclear.
Methods:  In the present study study, we investigated the therapeutic and prophylactic effects of protopanaxatriol saponin (PT) on testicular organ weight and morphology, testicular germ cells, proliferation, differentiation and spermatogenesis after induction of toxicity by a chemotherapeutic agent, busulfan, in male mice.
Results:  Intraperitoneally (IP) busulfan treatment markedly decreased the organ weight of testis, caput and cauda epididymis. After the treatment, the testes had collapsed seminiferous tubules with incomplete spermatogenesis. However, a single dose of busulfan treatment followed by PT injection showed milder damage on seminiferous tubules than busulfan alone.
Conclusion:  These results suggest that PT is effective in recovery of the male reproductive organ, and induced an increase in the number and viability of germ cells overcoming busulfan toxicity. PT might have applications in the recovery of male infertility arising from azoospermia and oligospermia.     

A percutaneous large-needle aspiration biopsy technique for histologic examination of the testis in infertile patients

OBJECTIVE: To describe a relatively new percutaneous large-needle aspiration biopsy technique for histologic examination of the testis in infertile patients. DESIGN: Retrospective analysis of clinical and pathologic data. SETTING: Clinical and academic research environment. PATIENT(S): Sixty-six infertile patients who underwent testicular biopsy. INTERVENTION(S): Local anesthesia was induced through spermatic cord block with lidocaine, and a relatively large needle (usually 18- or 20-gauge) was introduced percutaneously into the testicle without a scrotal incision. MAIN OUTCOME MEASURE(S): The number of seminiferous tubules per histologic section of each testicular biopsy sample. RESULT(S): A mean of 74 seminiferous tubules were obtained in the histologic sections of each biopsy sample. This number varied according to the size of the needle used; it was 24.7 when a 21-gauge needle was used, 56.2 when a 20-gauge needle was used, and 103 when an 18-gauge needle was used. The biopsies were performed in the office. No significant hematomas occurred, no antibiotic prophylaxis was prescribed, and no postbiopsy medical or pharmacologic interventions were required. CONCLUSION(S): Tissue specimens as large as those obtained with open surgical biopsy can be obtained from the testicles of infertile patients with the use of a percutaneous technique that is easier, less costly, and safer than any previously reported.     

Characterisation of a sperm coating auto-antigen reacting with antisperm antibodies of infertile males using monoclonal antibodies

Objective In a previous study a number of sperm-specific antigens were identified which reacted with antisperm antibodies from both infertile and vasovasostomised males. To investigate the localisation and distribution of these antigens and their role in male fertility, monoclonal antibodies were raised against them; immunoblotting techniques were used to select only those antibodies which competed with human antisperm antibodies for these human auto-antigens.
Design One antibody, NW21, reacted with an 18 kDa auto-antigen present on epididymal sperm but absent from testicular sperm. Immunohistochemical studies showed that the antigen is produced in small basal cells between the columnar epithelium of the corpus epididymis, passes up into the tubule and then coats sperm passing along the epididymis. Sperm stored in the cauda epididymis and ductus deferens stain strongly for this sperm coating glycoprotein.
Conclusions The localisation of this antigen supports the suggestion that auto-immune infertility may represent a response to epididymal rather than testicular sperm. Monoclonal antibodies raised to unique and immunologically accessible sperm coating proteins, produced in the epididymis rather than in the testis, would seem to present an excellent theoretical solution to male contraception.     

大鼠睾丸受氯化镉损伤后生精上皮恢复的观察

目的:探讨氯化镉(CdCl2)染毒大鼠致睾丸损伤后生精上皮能否恢复。方法:20只雄性SD大鼠随机分为对照组(A组)与3个CdCl2实验组(B、C、D组),每组5只动物。B-D组的动物分别腹腔注射CdCl21mg/kg,qd×14d。对照组腹腔注射等体积0.9%NaCl。染毒结束后,处死A和B组大鼠,C组经口服灌胃给予ZnCl2100mg/kg,qd×7d,D组不给予ZnCl2。C和D组染毒结束后再继续饲养2个月(恢复期),观察睾丸生精上皮的组织学改变,比较4组的异常曲细精管率。结果:B组部分曲细精管显示病理改变,B组的异常曲细精管率与A组的比较有显著升高(P<0.01)。恢复期后C和D组的曲细精管有明显恢复,大多数曲细精管显示正常曲细精管组织学图像,C和D组的异常曲细精管率与B组的比较均有显著降低(P<0.01)。C和D组之间的异常曲细精管率比较没有显著差异(P>0.05)。结论:在本实验条件下,CdCl2损伤后的大鼠睾丸,在自然状态下和给予ZnCl2,生精上皮均可得到显著恢复。     

睾丸的生殖病理学研究(Ⅰ)——40例无精症与少精症患者睾丸的生殖病理观察

本文提出了睾丸生殖病理双重诊断法,即同时作出睾丸病理诊断及睾丸生精障碍分级诊断,对各种病理类型的主要特征作了介绍。另又分析了睾丸各组织成分的病理变化发生率及病理生理意义。     

Scanning electron microscopic study on the shape of infertile seminiferous tubules: a hypothesis of pathogenesis of idiopathic male infertility

The shape of infertile seminiferous tubules was studied on testicular biopsied specimens from patients with idiopathic male infertility using the scanning electron microscope. A parallel study of the same sections was performed by ordinary light microscopy. Normal seminiferous tubules were revealed to be shaped like a weaving cylinder, whereas infertile seminiferous tubules were strangulated in various places, or appeared similar to a tail-like rope which became gradually more slender. These strangulated or slender parts of tubules seemed to correspond to the smaller tubules with thickened tubular walls that were recognized under the light microscope. It is suggested that these thickened tubular walls suppress spermatogenesis by a nutritional disturbance, and the strangulations of infertile tubules interfere with sperm transport by tubular blockage or germinal disorganization and interrupted contractions of the tubules.     

Varicocele is associated with abnormal retention of cytoplasmic droplets by human spermatozoa

OBJECTIVE: To determine whether varicocele is associated with retention of sperm cytoplasmic droplets in infertile men. DESIGN: Retrospective study.Setting: University infertility clinic. PATIENT(S): Nonazoospermic men with idiopathic (n = 69) and varicocele-associated infertility (n = 73), and 20 fertile controls presenting for vasectomy. INTERVENTION(S): None. MAIN OUTCOME MEASURES(S): Standard semen parameters and percentage of spermatozoa with cytoplasmic droplets on Papanicolaou smears. RESULT(S): No statistically significant differences were found between the fertile and infertile groups with respect to semen volume. Fertile controls had significantly greater mean percent sperm motility and normal morphology than infertile men. The mean percentage of sperm with residual cytoplasm was statistically significantly different in all three groups. Infertile men with varicocele had the highest percentage of sperm with cytoplasmic droplets, the next highest level being in men with idiopathic infertility and the lowest level in fertile controls (11.7 +/- 1.0, 8.1 +/- 0.9 and 3.2 +/- 0.4%, respectively, P<.0001). CONCLUSION(S): Our data show that idiopathic and even moreso, varicocele-related male infertility are conditions associated with impaired disposal of residual sperm cytoplasm by the testis and/or epididymis. These data provide a possible mechanism for the observed semen abnormalities and reduced fertility potential associated with varicocele and idiopathic male infertility.     

Effect of TSAA-291 on male reproductive tract and fertility of the rat

Adult male rats were injected with TSAA-291 (16 beta-ethyl-17 beta-hydroxyestr-4-en-3-one), a steroidal antiandrogen, daily for 30 days at two doses, viz., 10.0 and 25.0 mg/kg. The treatment caused a dose-dependent reduction in the weights of testis, epididymis, and other accessory sex glands. In the animals that received the high dose, 50% of the rats showed spermatogenic arrest in about 20% of the seminiferous tubules and Leydig cell morphology was normal. The levels of glycerylphosphorylcholine and sialic acid were significantly reduced in the cauda epididymis of all the treated rats. There was a reduction in the number of motile spermatozoa in the cauda epididymis of rats treated with high dose of the antiandrogen. This was accompanied by a reduced number of females exhibiting spermatozoa in their vaginal smears. A combination treatment of the antiandrogen and androgen appears to be a promising approach for fertility regulation in the male.     

Decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility

Objective: To examine the expression of the heat shock protein hsp70-2, and the possible relationship with the pathogenesis of male infertility.

Design: Prospective study.

Setting: Reproductive testing laboratory in a university hospital.

Patient(s): Men undergoing testicular biopsy during an investigation of subfertility.

Intervention(s): Testicular tissues were obtained from biopsies of men undergoing infertility evaluation and subdivided into three groups: normal testes, maturational arrest and Sertoli cell-only syndrome. Immunostaining and Western blotting techniques determined expression of the heat shock protein hsp70-2

Main Outcome Measure(s): Expression of the heat shock protein hsp70-2 in the testes.

Result(s): The experimental data demonstrated that the heat shock protein hsp70-2 was expressed in the normal and maturation arrest testicular specimens. The heat shock protein hsp70-2 was strongly present in the cytoplasm of spermatocytes and spermatides in the adluminal compartment of the seminiferous epithelium in normal testis. However, maturation arrest testis tissue demonstrated light staining in spermatocytes and spermatides, and Sertoli-only specimens demonstrated no staining for the heat shock protein hsp70-2. The Western blotting data showed a 70-kDa heat shock protein in the normal and maturation arrest testicular tissues, but not in the Sertoli-only tissues.

Conclusions: These results suggest that the heat shock protein hsp70-2 is expressed in spermatocytes and spermatides in normal and maturation arrest tissues. However, the expression of the heat shock protein hsp70-2 was low in maturation arrest, and no heat shock protein hsp70-2 was demonstrated in Sertoli-only specimens. Therefore the decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility.