实验性精索静脉曲张大鼠睾丸组织IL-1和NO的变化

目的 :研究精索静脉曲张 (VC)对大鼠睾丸组织中IL 1和NO含量的影响。　方法 :采用部分结扎左肾静脉及左髂总静脉之间的交通支 ,制备Wistar大鼠VC模型。将大鼠随机分为手术结扎组 (VC组 ,n =30 )和假手术组 (n =2 0 ) ,分别测定 2组大鼠睾丸组织中IL 1和NO的含量 ,并加以比较。　结果 :①IL 1、NO含量在左侧睾丸组织中VC组均显著高于假手术组 (P <0 .0 1) ,而在右侧睾丸组织中 2组比较差异无显著性 (P >0 .0 5 ) ;②IL 1与NO呈正相关关系 (r =0 .5 72 ,P <0 .0 1)。　结论 :VC时睾丸组织IL 1和NO的变化 ,可能是造成睾丸损伤、精子发生障碍甚至不育的原因     

茶多酚对精索静脉曲张大鼠睾丸生精细胞凋亡的影响

目的:研究茶多酚对精索静脉曲张大鼠睾丸生精细胞凋亡的影响,分析茶多酚对精索静脉曲张大鼠生精功能是否具有保护作用。方法:清洁级青春期Wistar大鼠32只随机分为4组,每组8只:假手术组(仅模拟手术过程,不结扎血管)、模型组(建立左精索静脉曲张模型)、茶多酚低剂量组[建立精索静脉曲张模型并给予10 mg/(kg·d)茶多酚干预]、茶多酚高剂量组[建立精索静脉曲张模型并给予40 mg/(kg·d)茶多酚干预]。建立左侧精索静脉曲张模型4周后,假手术组及模型组均给予生理盐水1 ml/100 g,茶多酚低剂量组及茶多酚高剂量组分别给予茶多酚10 mg/kg(用生理盐水配置成浓度为1 mg/ml)和40 mg/kg(用生理盐水配置成浓度为4 mg/ml),灌胃,1次/日,持续4周。4周后4组大鼠均处死并分别取左侧睾丸组织,检测低氧诱导因子-1(HIF-1)、Bcl-2、Bax、细胞色素C(Cyt C)和caspase-3在睾丸组织中的表达及生精细胞的凋亡并计算凋亡指数(AI)。结果:茶多酚干预组大鼠Bcl-2表达高于模型组(110.03±3.07),低于假手术组(154.18±2.96);茶多酚低剂量组大鼠Bcl-2(127.67±1.28)表达低于茶多酚高剂量组(136.41±1.95),差异有统计学意义(P0.05)。茶多酚干预组大鼠HIF-1、Bax、Cyt C和caspase-3的表达低于模型组,高于假手术组;茶多酚低剂量组大鼠HIF-1、Bax、Cyt C和caspase-3的表达高于茶多酚高剂量组,差异有统计学意义(P0.05)。假手术组AI最低,茶多酚干预组AI低于模型组,茶多酚高剂量组AI低于茶多酚低剂量组,差异有统计学意义(P0.05)。结论:茶多酚能显著降低精索静脉曲张大鼠睾丸生精细胞的凋亡。     

实验性精索静脉曲张对大鼠睾丸生精细胞凋亡的影响

目的 :探讨大鼠精索静脉曲张对睾丸生精细胞凋亡的影响 ,阐明精索静脉曲张引起不育的病理机制。　方法 :选用雄性SD大鼠 32只 ,随机分为假手术组、4 5d模型组、6 0d模型组及 90d模型组。采用肾静脉缩窄法建立精索静脉曲张动物模型 ,流式细胞术分别检测各组大鼠睾丸凋亡细胞数及各级生精细胞数。　结果 :假手术组未出现明显的凋亡峰 ,各模型组均出现明显的凋亡峰 ,且随着造模时间的延长凋亡峰增高。　结论 :精索静脉曲张可引起睾丸生精细胞大量凋亡 ,各级生精细胞数减少 ,这可能是精索静脉曲张引起睾丸生精功能障碍从而导致不育的根本机制     

Ⅲ度精索静脉曲张患者显微结扎术后生精功能的变化

目的：研究Ⅲ度精索静脉曲张显微结扎术后生精功能的变化,以探讨手术的价值.方法：选择Ⅲ度精索静脉曲张并男性不育患者118例,观察其显微结扎术前后睾丸体积、精子密度、活率、形态、血清卵泡刺激素（FSH）、睾酮（T）及抑制素B浓度的变化.结果：患者手术6个月后的左侧睾丸体积（P〈0.05）、精子密度（P〈0.01）及活率（P〈0.05）均明显提高,血清FSH水平明显降低（P〈0.05）,T（P〈0.01）及抑制素B（P〈0.05）水平明显提高.而睾丸体积及精子形态无显著变化.结论：显微结扎术能改善Ⅲ度精索静脉曲张患者的生精功能.     

精索静脉曲张大鼠睾丸低氧与生精细胞凋亡的研究

目的 观察实验精静索静咏曲张(varicocele,VC)大鼠睾丸低氧诱导因子1α(HIF-1α)的表达与睾丸生精细胞凋亡的关系,探讨VC导致不育的病理生理机制.方法 40只大鼠,随机分为3组.建立模型49d后,取其左侧睾丸组织,检测睾丸组织的HIF-1α的表达和生精细胞的凋亡率.结果 westernblot和免疫组化检测的实验组睾丸HIF-1α的表达(2.529±1847,78.57%)显著高于对照组(0.308±0.165,14.29%)和假手术组(0.309±0.164,0.00%)(P<0.05),差异均具统计学意义;实验组睾丸细胞凋亡指数(20.79±5.70)显著低于对照组(0.6±1.4)和假手术组(0.36±0.71)(P<0.001),差异均具统计学意义;实验组睾丸HIF-1α的表达与其细胞凋亡指数呈正相关(r=0.844,P=0.017).结论 VC可引起睾丸低氧,而低氧可以通过诱导生精细胞凋亡来引起睾丸功能的改变.同时,HIF-1α是一种预测生精细胞凋亡程度的有用指标.     

精子凋亡和生精细胞凋亡检测与精索静脉曲张的关系

精索静脉曲张(varicocele,VC)常人发病率在8%～20%,在男性不育症患者可达39%.协和医院统计1310例男性不育症中VC为466例,占35.57%[1].柯明辉(2010)统计中日友好医院不育症2599例,有VC者703例,占27.05%.VC是慢性渐进性对睾丸生殖功能有损伤的疾病,每增长1岁精液异常危险性提高0.023倍(2.3/100倍),是非VC精液异常危险率的1.2倍[2].发生于左侧睾丸者约为70%～90%,虽然发生在单侧,但它是一种双侧睾丸损伤性疾病,还具有累积性损害的特征,长时间的VC会引起睾丸功能下降,出现睾丸生殖功能障碍[3].青少年患者应该尽早治疗[4].本文就VC精子形态学、精子和生精细胞凋亡检出率与VC的关系综述如下.     

精索静脉曲张大鼠生精细胞凋亡的实验研究

目的 :探讨外科所致的精索静脉曲张对大鼠生精细胞凋亡的影响。　方法 :采用成年雄性Wistar大鼠复制左精索静脉曲张模型 ;用原位末端脱氧核苷酸转移酶标记法 (TUNEL)检测睾丸生精细胞凋亡。　结果 :实验组的细胞凋亡率显著高于假手术对照组 (P <0 .0 5 )。　结论 :外科所致的精索静脉曲张大鼠睾丸生长减缓 ,生精细胞凋亡增加 ,这可能是其影响生育能力的机制之一。     

精索静脉曲张导致睾丸生精细胞增殖与凋亡的变化

精索静脉曲张（VC）是男性不育的原因之一，在正常人群中VC的发生率为15％～17％，而不育者中VC的发生率占30％～40％。本研究旨在通过观察左侧VC患者双侧睾丸生殖细胞的增殖、凋亡的变化，探讨该病的病理生理机制。     

实验性精索静脉曲张大鼠生精细胞凋亡的周期时相性研究

精索静脉曲张(varicocele,VC)为泌尿外科常见疾病,其中约21%～41%系因不育而就诊,但截至目前精索静脉曲张导致不育的机制仍不十分清楚[1].由于生精细胞凋亡与不育关系密切,本研究应用流式细胞术(flow cytometry,FCM),分析实验性精索静脉曲张大鼠睾丸生精细胞凋亡的周期时相性,从细胞分子生物学角度探讨精索静脉曲张所致不育的病理机制.     

Impact of varicocelectomy on premature ejaculation in varicocele patients

In a prospective manner, we studied the effect of varicocelectomy on premature ejaculation and testicular hormonal function in patients with varicocele. Microsurgical subinguinal varicocelectomy was carried out in 73 patients with clinical varicocele associated with premature ejaculation (group 1) and compared with 56 patients without operative intervention (group 2; control). The primary efficacy variable was the mean change in the Premature Ejaculation Diagnostic Tool. The changes in International Index of Erectile Function questionnaire, total serum testosterone and testicular size were also assessed. In group 1, the Premature Ejaculation Diagnostic Tool decreased significantly after varicocelectomy, from 15.56 to 11.37 (P < 0.001), indicating improvement of premature ejaculation. Thirty patients (41.1%) showed improvement of premature ejaculation compared to 5.3% in the control group (P < 0.001). In group I, but not in group II, testosterone levels and International Index of Erectile Function values increased significantly when compared with pre‐operative values (P < 0.001 and 0.040 respectively). Testicular size increased after varicocelectomy, but this increase was nonsignificant till the end of study (P = 0.054). We concluded that varicocelectomy is clearly related to improvement of premature ejaculation and testicular hormonal function in varicocele patients.     

LM23基因沉默引起大鼠睾丸生精细胞凋亡

目的 研究LM23基因敲低后生精细胞的凋亡状况及与凋亡相关基因表达改变.方法 用TUNEL方法检测睾丸细胞的凋亡情况,用大鼠全基因组表达谱芯片检测LM23基因敲低后凋亡相关基因的表达改变.结果 TUNEL结果显示,LM23基因敲低侧大鼠睾丸生精小管内大量生精细胞发生凋亡.大鼠全基因组表达谱芯片分析结果显示,许多促凋亡基因如Bcl-2家族的促凋亡基因表达上调,而抗凋亡基因如Faf1和Zfp91基因的表达明显下调.结论 敲低大鼠的LM23基因,启动了Bcl-2家族介导的线粒体凋亡途径,引发了生精细胞发生大量凋亡.     

Study on the effect of glucocorticoid on apoptosis of rat spermatogenic cells

The effect of glucocoiticoid on the apoptosis of rat spermato-genic cells labeled with TUNEL was observed in vivo by laser con-focal microscopy and electron microscopy. The results showed thatlarge doses of glucocorticoid increased the apoptosis of rat spermto-genic cell. (Chin J Androl 2001; 2: 98-101)     

精索静脉曲张大鼠睾丸自噬对生精细胞的影响

目的:研究精索静脉曲张(VC)大鼠模型中睾丸不同水平自噬对生精细胞凋亡的影响。方法:雄性SD大鼠54只随机分为6组:空白对照组、雷帕霉素对照组、氯喹对照组各6只,VC组、VC+雷帕霉素组、VC+氯喹组各12只。采用HE染色观察睾丸、附睾组织形态学变化,并对睾丸及附睾中精子形成情况进行评分,TUNEL染色检测生精细胞凋亡指数(AI),Western印迹检测LC3、p62、Bax、Bcl-2的表达。结果:空白对照组、雷帕霉素对照组、氯喹对照组大鼠睾丸及附睾组织均未发生明显形态学变化,精子形成情况评分及AI亦无显著差异(P>0.05);VC组大鼠睾丸及附睾组织发生明显病理损伤,精子形成情况评分显著降低(P<0.01),AI显著升高(P<0.01),但VC+雷帕霉素组较VC组明显改善,而VC+氯喹组较VC组轻度加重。此外,与空白对照组比较,VC组自噬相关蛋白LC3(包括LC3-Ⅱ/LC3-Ⅰ的比值)和促凋亡蛋白Bax表达显著增加(P<0.01),抑凋亡蛋白Bcl-2表达则显著降低(P<0.01);且VC+雷帕霉素组LC3与Bcl-2表达显著高于VC组(P<0.01),p62和Bax表达则显著低于VC组(P<0.01);而VC+氯喹组LC3与Bcl-2表达显著低于VC组(P<0.01),p62和Bax的表达则显著高于VC组(P<0.01)。结论:VC可诱导大鼠睾丸自噬和生精细胞凋亡,上调自噬可抑制生精细胞凋亡,阻滞自噬则可促进生精细胞凋亡。     

Impact of varicocelectomy on urine dopamine value in patients with premature ejaculation and varicocele

Dopamine is an important regulator of male sexual function and behaviour. Decreased levels of this substance have been observed in blood and seminal plasma of infertile men. Hence, this study was carried out to determine the impact of varicocelectomy on 24‐hr urine dopamine values in patients with both premature ejaculation (PE) and varicocele. In this prospective study, 55 consecutive patients with premature ejaculation and grade 2 or 3 varicocele were enrolled. The urine dopamine level was measured in a 24‐hr sample by HPLC method in a single laboratory centre. Two samples were gathered from each patient, one before and the other 1 month after varicocelectomy. The mean initial and final 24‐hr urine dopamine levels were 259.6 ± 86.3 and 602.9 ± 88.4 micrograms, respectively, showing statistically significant increase (p = .0001), while there was no statistically significant difference between urine volume and creatinine. However, the change in intravaginal ejaculation latency time value was not statistically significant at this short time visit. According to our findings, varicocelectomy results in a significant increase in 24‐hr urine dopamine levels in patients with premature ejaculation and varicocele. However, the effect of this change on premature ejaculation in the long term has yet to be determined.     

Impact of varicocelectomy on the proteome profile of testicular tissues of rats with varicocele

Varicocele (VC) is a common cause of male infertility, but the molecular mechanisms involved in its pathogenesis are unknown. We investigated the impact of varicocelectomy (VCT) on proteome profiles in testicular tissues of rats with VC, and analysed associated target genes and signalling pathways. Sixty male rats with VC were divided into two groups: control (n = 30), and VCT (n = 30). Tissues were collected 4 weeks after sham or VCT surgery. Matrix‐assisted laser desorption/ionisation time‐of‐flight/time‐of‐flight mass spectrometry (MALDI‐TOF/TOF MS) was used to analyse the comparative proteome profiles. Kyoto Encyclopaedia of Genes and Genomes (KEGG) Orthology‐Based Annotation System was used for bioinformatic analysis. Fifteen proteins were differentially expressed between control and VCT groups. These differentially expressed proteins are associated with several specific cellular processes associated with the pathogenesis of testicular growth arrest associated with VC. Furthermore, the evaluation by transmission electron micrograph showed that VCT could decrease apoptosis of spermatogenic cells in rats. Understanding such molecular pathways might provide physicians with a better insight into VC and with potential targets for treatment.     

Dnajb13在隐睾小鼠睾丸中的表达及其与生精细胞凋亡的关系

目的:通过分析热休克蛋白40家族成员Dnajb13在成年小鼠生精细胞中的定位,研究其mRNA和蛋白在隐睾小鼠睾丸中表达量和定位的变化情况,初步探讨其在生精细胞凋亡中的作用。方法:手术建立单侧隐睾小鼠模型,采用免疫组化方法研究Dnajb13在小鼠生精细胞中的定位;采用TUNEL、流式细胞检测结合形态学观察确认小鼠隐睾模型是否建立成功,并分析生精细胞凋亡情况;采用qPCR、Western印迹以及免疫组化分析隐睾手术后第1、2、3、4、6、9和15天与正常侧相比隐睾侧Dnajb13 mRNA和蛋白的表达及定位变化情况。结果:免疫组化结果显示在成熟小鼠正常睾丸组织中,Dnajb13表达于生精上皮周期Ⅸ～ⅩⅥ期长形精子中;成功构建隐睾小鼠模型。TUNEL结合流式细胞检测表明I期圆形精子以及分裂期初级精母细胞对温度升高最敏感,隐睾模型手术后最早发生凋亡的生精细胞是圆形精子细胞(Ⅰ～Ⅷ期),从17.09%降为6.52%(P<0.05),随后是变形期精子细胞(Ⅸ～ⅩⅥ期),最后为初级精母细胞;qPCR结果显示术后第3天Dnajb13 mRNA表达量轻微上升,约为正常侧的1.6倍(P<0.05),第4天开始下降,约为正常侧的1.2倍;Western印迹结果显示术后第1～3天Dnajb13蛋白表达量无显著变化,第4天开始下降为正常侧的0.68倍(P<0.05),至第9天均急剧降下降为正常侧的0.4倍,Dnajb13在术后第6天已经凋亡的多核巨细胞中开始表达。结论:Dnajb13定位于成熟小鼠变形期精子细胞以及成熟精子尾部,参与了精子变形过程以及精子鞭毛的形成。隐睾状态下Dnajb13在凋亡的多核巨细胞中表达,可能参与了Ⅵ～Ⅷ阶段的圆形精子细胞的凋亡。     

精索静脉曲张大鼠睾丸瘦素表达的实验研究

目的研究精索静脉曲张对大鼠睾丸组织瘦素表达的影响，探讨精索静脉曲张引起不育的病理机制。方法选用雄性SD大鼠25只，随机分为模型组15只，假手术对照组10只。采用肾静脉缩窄法建立精索静脉曲张动物模型，半定量RT-PCR法分别检测两组大鼠睾丸瘦素mRNA含量。结果精索静脉曲张模型组和假手术对照组大鼠睾丸均有瘦素表达，模型组较假手术对照组瘦素mRNA含量显著增加（P〈0.05）。结论精索静脉曲张可引起睾丸瘦素mRNA表达显著增加，这可能是精索静脉曲张引起睾丸生精功能障碍导致不育的重要机制之一。