交变磁场照射对小鼠睾丸生殖功能的影响

目的:探讨交变磁场的物理作用与生物学效应的关系,研究磁场对小鼠睾丸生殖功能的影响。方法:30只ICR雄性小鼠随机均分为正常对照组、X线照射组、弱磁场(1000Hz)组、强磁场(2000Hz)1h组和强磁场2h组。照射后7d处死小鼠,分析附睾精子活率,对睾丸组织切片行HE染色观察睾丸病理组织变化,以Johnsen评分标准,评定睾丸变化积分。结果:精子活率正常对照组为(42.37±10.24)%,X线照射组为(39.00±12.35)%,弱磁场组为(36.00±17.28)%,强磁场1h组为(10.72±5.67)%,强磁场2h组为(4.44±2.87)%,强磁场1h组和2h组与正常对照组比较,精子活率明显下降(P<0.01)。睾丸病理变化Johnsen评分随着交变磁场的加大和时间延长积分越低,睾丸损伤越严重。结论:强、弱磁场均引起小鼠睾丸功能破坏,导致生精小管损伤,间质、间质细胞损坏,基膜增厚,腔内生精细胞排列紊乱,凋亡、坏死和脱落增多,导致无精子发生。     

红景天提取物对顺铂诱导小鼠睾丸支持细胞损伤的保护作用

目的:体外研究红景天提取物对顺铂(cDDP)诱导小鼠睾丸支持细胞(TM4)损伤的保护作用,并探讨其可能的作用机制。方法:体外培养正常TM4细胞,将细胞分组,MTT法测定红景天提取物、cDDP分别对TM4细胞生长的影响以及红景天提取物对cDDP诱导TM4细胞损伤的保护作用,同时观察细胞形态学变化。硫代巴比妥酸法测定细胞丙二醛(MDA)含量,黄嘌呤氧化酶法测定总超氧化物歧化酶(T-SOD)含量,二硫代二硝基苯甲酸法测定谷胱甘肽(GSH)含量。结果:MTT结果显示,红景天提取物在0.0125².5 mg/L终质量浓度范围内可拮抗cDDP诱导TM4细胞增殖率的降低(P<0.01),改善cDDP诱导下TM4细胞形态学变化,减少胞体减小、变圆,细胞脱壁等现象。抗氧化实验结果表明,0.0147 g/L的cDDP诱导下的TM4细胞内MDA含量[(3.63±0.02)nmol/mgprot]较正常对照组[(2.15±0.02)nmol/mgprot]显著升高(P<0.01),T-SOD、GSH含量[(6.57±0.05)U/mgprot、(1.42±0.06)mg/gprot]较正常对照组[(10.86±0.02 U/mgprot、(2.59±0.05)mg/gprot]显著降低(P<0.01);0.1 mg/L红景天提取物可显著降低(P<0.01)cDDP诱导TM4细胞内MDA含量[(1.94±0.00)nmol/mgprot]、减缓T-SOD[(8.50±0.02)U/mgprot]和GSH[(2.41±0.04)mg/gprot]耗竭。结论:红景天提取物能有效抑制cDDP诱导TM4细胞损伤,其机制可能与红景天的抗氧化能力有关。     

急性弓形虫感染对雄性小鼠睾丸生精功能影响的初步研究

目的:探讨弓形虫急性感染对雄性小鼠睾丸生精功能的影响。方法:将26只成年雄性小鼠随机均分为感染组和正常对照组;感染组腹腔注射0.3ml1×103/ml弓形虫速殖子,感染小鼠睾丸,正常组腹腔注射等量生理盐水,两组睾丸制成直接印片及病理切片,观察小鼠生精细胞的病理变化及弓形虫侵入生精细胞情况。同时比较两组睾丸乳酸脱氢酶同工酶、精子密度、精子活动率、精子畸形率。结果:感染组睾丸乳酸脱氢酶同工酶、精子密度、精子活动率、精子畸形率分别为53.19±18.04、(15.01±2.42)×106/ml、(8.26±2.57)%、(17.69±11.91)%,正常组为68.71±17.79、(23.87±6.66)×106/ml、(13.21±2.82)%、(11.30±6.60)%,两组比较差异均有显著性(P<0.05)。结论:弓形虫感染对雄性小鼠睾丸生精功能有一定的影响。     

Diazepam administration prevents testosterone decrease and lipofuscin accumulation in testis of mouse exposed to chronic noise stress

Lipofuscin is an autofluorescent and undegradable material, which accumulates in tissues during ageing and under different types of stress. Among these, oxidative stress represents a major trigger for lipofuscin formation. However, prolonged noise exposure is also an effective stressful stimuli. Diazepam may inhibit lipofuscinogenesis in liver and prevent the noise-induced reduction of the steroidogenesis in the adrenal gland. The aim of the study was to ascertain whether chronic noise exposure causes lipofuscin accumulation in mouse testis, and to evaluate the effects of diazepam administration. Eight-week old mice were either exposed for 6 weeks (6 h day(-1)) to white-noise (group A), or received diazepam (3 mg kg(-1), i.p.) before noise exposures (group B), while a further group was used as control (group C). Light fluorescence and transmission electron microscopy revealed lipofuscin in large amounts in the Leydig cells in mice of group A, which concomitantly had low serum testosterone levels; pre-treatment with diazepam occluded both effects. The present study indicates that: (i) chronic noise exposure causes lipofuscin accumulation at the level of the Leydig cells and a decrease in testosterone; (ii) all these effects are suppressed by pre-treatment with diazepam. As the Leydig cells represent the only cellular type of the interstitial testicular tissue having peripheral benzodiazepine receptors, these results could be explained by the capacity of the peripheral benzodiazepine receptors to prevent reactive oxygen species damage and to increase the resistance of these cells to oxidative stress.     

GATA-4在小鼠睾丸中的表达定位研究

目的:观察小鼠睾丸内GATA-4免疫反应产物的特征与分布。方法:选取刚出生、2、4、6周雄性B6SJLF1/J小鼠各6只,取睾丸进行石蜡切片,应用免疫组化ABC法,DAB显色,观察GATA-4在不同时期睾丸中的表达情况。结果:GATA-4免疫反应产物在以上4个阶段小鼠睾丸Sertoli细胞、Leydig细胞均有分布,且4、6周组小鼠睾丸Leydig细胞阳性率比刚出生和2周组小鼠更高(P<0.01);但4、6周组小鼠,除以上两种细胞外,生精细胞中也有分布,6周组小鼠比4周组小鼠阳性细胞率高(P<0.01)。结论:小鼠睾丸内存在GATA-4,为研究GATA-4在睾丸性别决定和分化以及激素调控方面提供了形态学依据。     

Histologic changes in the mouse testis after bilateral vasectomy

Aim: To study the effect of vasectomy on histological appearance of the testis. Methods: Parkes strain mice were used as the animal model; they were bilaterally vasectomized (Vx) or sham-operated (So) and killed at intervals of 4, 6, 9, and 12 months after the operation. Testes were excised from 5 Vx and 5 So mice at each interval and processed for histological examination. Results: Testes of So mice showed normal histological features. By contrast, marked alterations were observed in the seminiferous tubules in testes of Vx mice, except in those killed 4 months after the operation. The seminiferous epithelium in the tubules was only 2 - 3 layers thick and showed much depletion of germ cells; in severe cases, the epithelium consisted of only a thin layer of Sertoli cells, spermatogonia and a few spermatocytes. Exfoliation of germ cells, occurrence of multinucleated giant cells and vacuolated appearance of the epithelium were of common features in the tubules. Furthermore, lumen of the rete testis in Vx mice was greatly dilated and showed accumulation of spermatozoa with immature germ cells; in mice vasectomized for 6 - 12 months, several macrophages ingesting spermatozoa were often observed in the lumen of the rete testis. Spermatic granuloma was also sometimes noticed in corpus or in cauda regions of the epididymis in mice vasectomized for 6 - 12 months. Conclusion: We suggest that consequences of vasectomy should be thoroughly understood in order to make this method rather more popular as a reversible method of male contraception.     

Osteotesticular protein tyrosine phosphatase expression in rodent testis

PURPOSE: In the last decade the novel receptor-like protein tyrosine phosphatase was identified and termed osteotesticular tyrosine phosphatase (OST-PTP) due to its restricted expression in bone and testis. OST-PTP expression is regulated during osteoblast differentiation and it shows stage specific expression in the testis. Confined OST-PTP expression in the basal compartment of the seminiferous tubules suggests that this protein may be a good candidate for a rodent germ stem cell marker. To test this hypothesis we determined the exact pattern of OST-PTP expression in the rodent testis. MATERIALS AND METHODS: Adult mouse and rat paraffinized testis sections were subjected to in situ hybridization using riboprobes against the receptor and catalytic domains of the protein OST-PTP. RESULTS: OST-PTP testicular expression in rodents is not confined to the spermatogonia, as previously suggested, but is also present in Sertoli cells in a stage independent pattern. This finding excludes the hypothesis that OST-PTP is a germ stem cell identification marker in rodents. In addition, we report the identification of a testicular OST-PTP isoform lacking part of a catalytic domain that is widely expressed during spermatogenesis in all cell types. CONCLUSIONS: This finding implies tight control over OST-PTP expression in the testis, which in turn suggests an important role for OST-PTP and its isoform in male germ cell differentiation.     

Effects of plants and plant products on the testis

For centuries, plants and plant-based products have been used as a valuable and safe natural source of medicines for treating various ailments. The therapeutic potential of most of these plants could be ascribed to their anticancer, antidiabetic, hepatoprotective, cardioprotective, antispasmodic, analgesic and various other pharmacological properties. However, several commonly used plants have been reported to adversely affect male reproductive functions in wildlife and humans. The effects observed with most of the plant and plant-based products have been attributed to the antispermatogenic and/or antisteroidogenic properties of one or more active ingredients. This review discusses the detrimental effects of some of the commonly used plants on various target cells in the testis. A deeper insight into the molecular mechanisms of action of these natural compounds could pave the way for developing therapeutic strategies against their toxicity.     

冷冻保存未成熟睾丸组织移植后生精细胞的生长发育木

目的采用睾丸组织移植模型,探讨通过简单易行的冷冻保存程序保存未成熟睾丸组织,并使冷冻后的组织恢复生精过程的可能性,为该动物模型应用在保存雄性生殖细胞、帮助癌症患者保存生殖能力以及保存濒危物种等方面提供进一步的实验依据。方法用DMEM培养基加入二甲基亚砜配制冷冻保存液,将新生1～2d昆明小鼠睾丸组织按分步冷冻的方法进行冷冻,最终放入-80℃超低温冰箱中保存起来。2周后取出冻存的睾丸组织,移植到雄性去势免疫缺陷小鼠背部皮下,分别于移植后4周、5周和8周取材,制作HE染色切片,观察移植物中生精小管结构和生精细胞的组成,并与新鲜移植的睾丸组织进行对比分析。结果采用实验中的冷冻程序保存的新生小鼠睾丸组织,在冷冻保存一段时间后再移植,其表现与新鲜睾丸组织移植相同,其中末成熟的生精细胞可以在受体中继续生长发育,并完成整个生精过程,发育成为精子。结论本实验中所采用的睾丸组织冷冻保存方法具有不需特殊设备、简便易行、适用范围广泛等优点,适用于多种场合中生精细胞的冷冻保存。     

趋化素样因子超家族在小鼠睾丸中的基因发育表达调控和蛋白表达定位

目的研究CKLFSF2小鼠同源序列cklfsf26基因在睾丸组织发育中的表达和定位,为其功能研究做出提示。方法利用RT-PCR方法检测cklfsf26基因在不同周龄小鼠睾丸组织中的表达情况；制备cklfsf2b多克隆抗体,利用免疫组织化学方法,检测cklfsf2b蛋白在小鼠睾丸组织中的表达定位。结果cklfsf2bmRNA在出生后第2周内出现,表达量逐渐增加,在成年前即达高峰并持续表达。cklfsf2b蛋白特异性的表达于睾丸Leydig细胞和Sertoli细胞,阳性信号均位于胞质内。结论cklfsf2b基因存在发育的表达调控,随着睾丸发育的日趋成熟,表达量逐渐增加：在Leydig细胞和Sertoli细胞的特异性表达表明其在睾酮合成和精子发生中均可能发挥作用,但其具体功能还需深入研究。     

枸杞多糖对体外培养小鼠睾丸组织形态学影响的研究

本研究是通过对小鼠睾丸组织培养，观察枸杞多糖（LBP）对不同培养条件下的睾丸组织形态学变化的影响。结果显示，32℃条件下培养的睾丸组织结构变化在对照组与LBP实验组之间差异不显著；37℃条件下培养48h时组间差异不显著，但在培养72h时，对照组显示曲精小管上皮细胞结构紊乱，大量细胞脱落和退化，管壁上仅剩精原细胞和支持细胞；在LBP实验组，曲精小管上皮细胞结构明显好于对照组，生精上应仍为复层上皮，管腔内脱落细胞较少。此结果表明，LBP可减弱由于培养温度过高所致的曲精小管结构损伤。     

Effects of testosterone and vitamin E on the antioxidant system in rabbit testis

The aim of the study was to investigate the effects of testosterone propionate and vitamin E on the antioxidant system in the testis. Thirty-two male New Zealand White rabbits were randomly divided into four groups. The first group was used as control. The second group was injected with testosterone propionate, the third group vitamin E and the fourth group vitamin E and testosterone propionate combination. All treatments were carried out during 6 weeks and oxidative parameters were evaluated in homogenized testicular tissue. The levels of vitamin E and the activity of glutathione peroxidase were lower (P < 0.05) in the testosterone group than in controls. However, vitamin C and malondialdehyde levels were higher (P < 0.05) in this group than in controls. The levels of reduced glutathione, beta-carotene, vitamin C and E increased, but malondialdehyde levels decreased in the vitamin E group, when compared with controls (P < 0.05). Vitamin E and beta-carotene levels were significantly higher (P < 0.05) in the combination group than in testosterone group. However, MDA levels were lower (P < 0.05) in combination group than in the testosterone group. In conclusion, administration of testosterone propionate led to a significant elevation of oxidative stress. Vitamin E is quite an effective antioxidant which protects rabbit testis against lipid peroxidation, and, testosterone-induced lipid peroxidation could be improved by additional vitamin E treatment.