低氧对大鼠精子体外顶体反应和顶体酶、透明质酸酶活性的影响

目的研究低氧对大鼠精子顶体酶、透明质酸酶活性的影响以及对精子体外顶体反应发生的影响。方法成年Wistar大鼠随机分为4个组:常氧对照组、低氧5d组、低氧15d组和低氧30d组。低氧组置低压舱内模拟高原5000m,分别低氧5、15和30d。采用明胶薄膜法、透明质酸底物转化法、溶血磷脂胆碱(LPC)诱导金霉素荧光染色法,测定低氧对附睾尾精子顶体酶、透明质酸酶活性和体外顶体反应发生率的影响。结果低氧5、15和30d组大鼠精子顶体反应发生率,与常氧对照组相比均显著降低,分别由常氧对照组的(30.5±3.5)%,下降至(11.7±0.9)%(P<0.01)、(10.8±1.0)%(P<0.01)和(10.0±1.4)%(P<0.01);低氧5、15和30d组大鼠精子顶体酶阳性率降低,分别由常氧对照组的(81.67±7.16)%,下降至(54.17±3.82)%(P<0.01)、(30.00±3.92)%(P<0.01)和(43.00±3.63)%(P<0.01);低氧各组透明质酸酶活性无显著改变(P>0.05)。结论低氧抑制了精子顶体酶活性和顶体反应发生。     

阉割对大鼠颌下腺雄激素受体及表皮生长因子的影响

目的:观察阉割对大鼠颌下腺雄激素受体(AR)及表皮生长因子的影响。方法:60只30~60日龄的W istar雄性大鼠,随机分为阉割组、假手术组和正常对照组,每组20只。1周后摘除各组大鼠颌下腺。颌下腺匀浆后用于W estern印迹分析。结果:与其他两组相比,阉割组颌下腺中的AR含量明显降低(P<0.05),而且,颌下腺分泌的表皮生长因子亦低于其他两组(P<0.05)。结论:阉割影响了颌下腺中AR的产生,并进一步影响了颌下腺分泌表皮生长因子的功能。     

肝细胞生长因子对大鼠颌下腺细胞增殖的影响

目的　研究肝细胞生长因子 (HGF)对体外培养大鼠颌下腺细胞增殖的剂量 效应及时间 效应。方法　采用体外颌下腺细胞培养在含 1 0 %胎牛血清的合成培养液 (DMEM) ,实验组分别加入不同浓度的HGF ,利用噻唑蓝 (MTT)比色法检测细胞的增殖能力 ,研究HGF对颌下腺细胞增殖的影响。结果　在 1 0 %胎牛血清配制的DMEM含HGF(1 .0～ 1 0 0 0 .0 μg/L)可促进培养细胞的增殖 (P <0 .0 1 ) ,且在一定浓度范围内HGF(1 .0、5 .0、1 0 .0、50 .0、1 0 0 .0 μg/L)呈剂量 效应关系 ,最大效应剂量为 1 0 0 .0 μg/L。加入HGF第 3天开始显示出明显的促增殖效应。 结论　HGF在 1 0 %胎牛血清DMEM培养条件下能够促进颌下腺细胞的增殖和分化 ,经体外条件培养的颌下腺细胞可以作为组织工程学研究中的细胞来源     

抗精子抗体对精子顶体酶活性的影响

本文旨在观察抗精子抗体对精子顶体酶活性的影响。选择男性不育者５０例与正常生育者２０例，采用固相酶染色法测抗精子抗体，以固定明胶薄膜法测精子顶体酶活性。结果发现５０例不育者抗精子抗体阳性率为５２％，不育者精子顶体酶活性明显低于生育者；抗精子抗体阳性者顶体酶活性明显低于阴性者，表明抗精子抗体可降低精子顶体酶活性。     

人绒毛膜促性腺激素对小鼠睾丸、附睾表皮生长因子和睾酮的影响

为了探讨人绒毛膜促性腺激素 (h CG)与颌下腺、睾丸和附睾表皮生长因子(EGF)的相互调节机制 ,本研究对 ICR雄性小鼠在切除颌下腺和给予 h CG前后应用放射免疫方法测定睾丸和附睾 EGF变化以及睾丸和血清睾酮 (T)变化。结果 :去颌下腺后 ,睾丸 EGF和血清 T不降低 (P>0 .0 5 ) ,附睾 EGF明显增加 (P<0 .0 5 ) ,睾丸 T显著降低(P<0 .0 5 ) ;去颌下腺给药组与去颌下腺组相比 ,睾丸和附睾 EGF明显增加 (P<0 .0 5 ) ,睾丸 T明显增加 (P<0 .0 5 ) ,血清 T显著降低 (P<0 .0 5 )。假手术给药组与假手术组相比 ,睾丸和附睾 EGF明显增加 (P<0 .0 5 ) ,睾丸 T明显增高 (P<0 .0 5 ) ,血清 T不增加 (P<0 .0 5 )。假手术给药组与去颌下腺给药组相比 ,睾丸 EGF明显增加 (P<0 .0 5 ) ,附睾EGF明显降低 (P<0 .0 5 ) ,睾丸 T和血清 T均明显增加 (P<0 .0 5 )。结论 :(1 )睾丸与附睾均能产生 EGF。 (2 ) h CG可通过调节睾丸 T生物合成影响睾丸、附睾 EGF含量。 (3)颌下腺促进睾丸 T的生物合成。 (4 )颌下腺和 h CG调节睾丸、附睾 EGF合成的机制不同。     

抗精子抗体对精子顶体酶活性的影响

目的；观察抗精子抗体对精子顶体酶活性的影响。方法：选择男性不育者５０例，与正常生育者２０例。采用固相酶染色法测搞精子抗体，固定明胶薄膜法测精子顶体酶活性。结果：５０例不育者抗精子抗体阳性率为５２５，不育者精子顶体酶生明显低于生育者；抗精子抗体阳怀者顶体酶活性低于阴性者。结论：抗精子抗体可降低精子顶体酶活性。     

抗精子抗体对精子顶体酶活性的影响

目的：观察抗精子抗体对精子顶体酶活性的影响。方法：选择男性不育者５０例，与正常生育者２０例。采用固相酶染色法测抗精子抗体，固定明胶薄膜法测精子顶体酶活性。结果：５０例不育者抗精子抗体阳性率为５２％。不育者精子顶体酶活性明显低于生育者；抗精子抗体阳性者顶体酶活性低于阴性者。结论：抗精子抗体可降低精子顶体酶活性。     

抗精子抗体对精子顶体酶活性的影响

本文旨在观察抗精子抗体对精子顶体酶活性的影响。选择男性不育者50例与正常生育者20例，采用固相酶染色法测抗精子抗体，以固定明胶薄膜法测精子顶体酶活性。结果发现50例不育者抗精子抗体阳性率为52％。不育者精子顶体酶活性明显低于生育者；抗精子抗体阳性者须体酶活性明显低于阴性者，表明抗精子抗体可降低精子顶体酶活性。     

IL-2对精子顶体酶及一氧化氮合酶活性的影响

目的　探讨白细胞介素 2对正常人及白细胞精子症精子功能的影响。方法　精子顶体酶的活性检测采用BAEE/ADH联合法。结果　白细胞精子症及正常人精子顶体酶的活性分别为 (2 7.6 8± 10 .0 6mU/ml,n =18;4 3.6 3± 7.6 1mU/ml,n =18) (P <0 .0 1) ;白细胞精子症及正常人精子一氧化氮合酶分别为 (2 .74± 0 .39U/mg pr,n =18;2 .74± 0 .39U/mgpr,n =18) (P <0 .0 1)。结论　白细胞介素 2对精子顶体酶和一氧化氮合酶均有明显抑制作用 ,提示这可能是导致精子受精能力下降的原因。     

Percoll优选对形态正常精子百分率和顶体酶活性的影响

目的 :了解形态正常精子百分率和顶体酶活性在Percoll优选前后的变化。　方法 :30例不育男性精液标本 ,应用计算机精液分析仪进行精子形态分析 ,分光光度比色法检测顶体酶活性。　结果 :Percoll优选后形态正常精子百分率和精子顶体酶活性均明显高于Percoll优选前 (P均 <0 .0 0 1)。　结论 :Percoll法优选精子后形态正常精子百分率和顶体酶活性增加。     

表皮生长因子对颌下腺细胞在胶原海绵黏附的影响

目的　探讨表皮生长因子 (EGF)对颌下腺细胞在胶原海绵支架上黏附的影响。方法体外原代培养SD鼠颌下腺细胞 ,并传至第 2代 ,将其分别接种于含表皮生长因子修饰的胶原海绵上 ,和不含表皮生长因子修饰的胶原海绵上。于接种后 7d取材 ,进行细胞计数计算细胞贴附率 ;苏木素 伊红 (HE)染色 ,扫描电镜观察等检测。结果　实验组含有EGF ,细胞贴附率为 73 .1% ,对照组不含EGF ,贴附率为 62 .8% (P <0 .0 5 ) ,HE染色及扫描电镜观察也证实 :实验组的颌下腺细胞在胶原海绵支架上黏附 ,增殖及分化均优于对照组。结论　EGF对颌下腺细胞在胶原海绵上的黏附有促进作用。     

不同浓度曲拉通X-100对制备颌下腺脱细胞基质的影响

目的探讨制备颌下腺脱细胞基质生物衍生支架材料过程中曲拉通X-100的最佳浓度。方法用3组不同浓度的曲拉通X-100试剂对30个颌下腺腺体进行脱细胞处理，并通过光镜、透射电镜及扫描电镜观察。结果0．5％和1％曲拉通X-100组脱细胞后，光镜下见大部分细胞轮廓存在，呈拥挤状态。透射电镜下见细胞膜破损，细胞器破碎，形成多个散在的碎块。扫描电镜下见细胞膜被破坏，表面呈丰窝状改变。而3．0％曲拉通X-100组脱细胞后，光镜下见颌下腺细胞成分完全消失，胶原纤维呈网状排列。透射电镜下见细胞器及细胞核消失，只见残留的细胞轮廓。扫描电镜下见细胞完全消失，只残留空虚的陷窝，基质胶原纤维粗细不等，相互交错呈网状结构。结论在制备颌下腺脱细胞基质时曲拉通X-100试剂的最佳浓度是3．0％。     

睾酮在精子生成过程中分泌动力学的实验研究

目的　研究睾酮在精子生成过程中的作用浓度。　方法　对ＳＤ 大鼠的精索静脉分别作双、单侧高位结扎后３、２１ 天睾丸间液及不同部位血清的睾酮浓度进行测定,并观察睾丸显微结构的变化。　结果　大鼠体内睾丸间液睾酮浓度最高;血清睾酮浓度依次为睾丸静脉［（４２－５０３ ±１２－７４９） ｍｇ／Ｌ］ 、近端精索静脉［（４２－５０３±１２ ．７４９） ｍｇ／Ｌ］ 、睾丸动脉［（５－５９８ ±３－６４９）ｍｇ／Ｌ］ 、尾静脉［（２．５３３±１－７１９） ｍｇ／Ｌ］和下腔静脉血［（２－４１８ ±１－４９５） ｍｇ／Ｌ］ 。双侧精索内静脉结扎后３ 天,双侧睾丸重量、血清和睾丸间液睾酮浓度均显著下降。左侧精索内静脉结扎后３ 天,左侧睾丸重量、睾丸动脉和睾丸间液睾酮浓度均显著下降;其余在正常水平。所有结扎侧睾丸精曲小管上皮轻度变性,并伴有部分结构不清。２１ 天后均恢复正常。　结论睾酮在体内除了正常循环外,还可能存在１ 个从睾丸睾丸静脉精索静脉精索动脉睾丸动脉睾丸的“小循环”。近端精索静脉的结扎对睾丸内的睾酮浓度和睾丸结构的影响是暂时性的,能自行恢复和修复     

Regional distribution of epidermal growth factor,testosterone and dihydrotestosterone in benign prostatic hyperplasia tissue

In benign prostatic hyperplasia (BPH), basic fibroblast growth factor (bFGF) is found to have a regional distribution, with concentrations in the periurethral zone (where the primitive fibrostromal nodule originates) higher than those of the peripheral subcapsular zone. The aim of the present investigation was to verify whether androgens and epidermal growth factor (EGF) are uniformly distributed from the periurethral to the peripheral zone or whether they show regional differences. Tissue samples, removed by transvesical resection from nine untreated BPH patients, sectioned in periurethral, subcapsular, and intermediate zones, were examined. In the periurethral zone, dihydrotestosterone (DHT), testosterone, and EGF, determined by radioimmunoassay (RIA) techniques after purification on Celite microcolumns and Sep-pak C18 cartridge, showed values significantly higher (mean ± SD: 1121±482 pg, 250±129 pg, and 6.89±3.28 ng/mg DNA, respectively;P<0.01) than those of the subcapsular zone (489±190 pg, 114±70 pg, and 3.40±1.90 ng/mg DNA, respectively). A positive linear correlation between EGF, testosterone, and DHT was also observed. The regional distribution of EGF, testosterone, and DHT was similar to that found for bFGF: the highest levels of these factors in the periurethral region allow us to hypothesize on their possible involvement in the rewakening of mesenchymal tissue, leading to the formation of the primitive fibrostromal nodule and then to BPH development.     

The effect of melatonin implants on blood testosterone and acrosin activity in spermatozoa of the ram

In a series of consecutive blood sampling in 15 days intervals over 15 weeks after implantation of melatonin in rams an increased mean value, basal level and number of peaks of testosterone was observed in samples of the third fortnight (45th day). This increase was greater in the autumn (breeding season) than in spring (non-breeding season). Total acrosin activity in spermatozoa was increased between days 35-56 (autumn) and days 49-70 (spring) after implantation and the relative increase was higher in autumn than in spring. The increase of acrosin activity was independent of the changes of testosterone. An increase of acrosin activity by melatonin, in cases of low activity, might improve fertilization rates in sheep not only during the breeding season, but also during the non-breeding season (after oestrus induction).     

荧光定量聚合酶链反应检测去颌下腺大鼠睾丸Bcl 2和Bax mRNA的改变

目的观察切除颌下腺对大鼠睾丸Bax和Bcl 2 mRNA表达的影响。方法切除大鼠颌下腺,分别于术后14、28和42 d处死大鼠,提取睾丸总RNA,反转录,设计特异性引物和Taqman探针,荧光定量聚合酶链反应(PCR)检测Bax和Bcl 2 mRNA的改变。结果随着颌下腺切除时间的延长,实验组大鼠睾丸Bax和Bcl 2 mRNA的比值显著升高,与对照组比较,有显著性差异(P<0.01)。结论颌下腺切除大鼠睾丸Bax和Bcl 2 mRNA比值明显升高,提示颌下腺切除所致的大鼠睾丸生精细胞的凋亡可能是由Bax和Bcl 2介导的线粒体途径进行,但Fas/FasL是否参与该凋亡过程还有待进一步研究。     

Effect of testosterone deprivation on expression of the androgen receptor in rat prostate, epididymis and testis

Adult rats were treated with ethane dimethane sulphonate (EDS) to eliminate the Leydig cells. This treatment resulted in very low levels of testosterone in the blood and in the testis. Furthermore, histological evaluation of spermatogenesis showed no marked differences between control and EDS-treated animals. In the ventral prostate, 5 days after EDS-treatment, a 4.0 +/- 0.3-fold up-regulation of androgen receptor (AR) mRNA was observed, together with a 2.2 +/- 0.2-fold increase in actin mRNA. In the epididymis, a 2.0 +/- 0.5-fold increase in AR mRNA level was observed, without a change in actin mRNA level. In the testes of EDS-treated rats, the AR mRNA level was not changed (1.02 +/- 0.17-fold of controls), and there was also no change in actin mRNA level at 5 days after EDS-treatment. These results indicate that AR mRNA expression in the ventral prostate and epididymis is regulated differentially by testosterone when compared to regulation in the testis. Testicular androgen binding sites were assayed by Scatchard analysis of the binding of 3H-R1881 to a nuclear fraction, that was isolated by a method which involved the use of liquid nitrogen and high sucrose buffer. The number of specific binding sites per testis in EDS-treated rats with testosterone-implants, remained unaltered compared to control rats (9.1 +/- 1.4 pmol/testis). In these rats, 20% of the normal testicular testosterone level was sufficient to maintain the androgen receptor in a tight nuclear binding (transformed) form. In testes from EDS-treated rats without testosterone-implants, the AR did not fractionate into the nuclear fraction; however, the total testicular AR content in these animals was close to control levels, as measured by nuclear 3H-R1881 binding after receptor transformation through injection of a high dose of testosterone (10 mg) 2 h before killing the rats (testosterone pulse). In the different experimental groups, FSH was not required to maintain the total testicular AR content (ligand binding). Immunoprecipitation and Western blotting of the testicular AR using specific monoclonal and polyclonal antibodies indicated that the total testicular amount of immunodetectable AR protein in long-term testosterone deprived rats was very low when compared to that in control rats or rats with testosterone-implants. This is in disagreement with results obtained in the ligand binding assay, and may point to a structural modification of the AR in the testis that possibly occurs in the prolonged absence of androgens.     

实验性精索静脉曲张对大鼠前列腺组织中睾酮水平及雄激素受体表达的影响

目的探讨实验性精索静脉曲张对大鼠前列腺组织中睾酮水平及雄激素受体表达的影响。方法将36只SD三级雄性大鼠随机分为实验组与对照组,每组18只,实验组建立精索静脉曲张模型,对照组接受假手术,于建模成功后第6、12和18周(各3个阶段,每阶段6只)采用放射免疫法测定血清中睾酮的含量、前列腺组织中睾酮的含量,并进行常规HE染色,观察前列腺组织上皮与基质的形态学变化,同时使用免疫组化法测定前列腺组织中雄激素受体的表达。结果实验组6周组、12周组、18周组血清睾酮、前列腺组织中睾酮水平明显低于同期对照组(P<0.05),且实验组12周组血清睾酮、前列腺组织中睾酮水平明显低于实验组6周组(P<0.05),实验组18周组血清睾酮、前列腺组织中睾酮水平明显低于实验组6周组、实验组12周组(P<0.05)。对照组前列腺组织正常,实验组随着时间的延长前列腺组织损伤越来越严重。实验组6周组前列腺组织中雄激素受体表达较同期对照组明显增强(P<0.05),实验组12周组、实验组18周组前列腺组织中雄激素受体表达较同期对照组与实验组6周组明显减弱(P<0.05)。结论实验性精索静脉曲张可降低大鼠前列腺组织中睾酮水平,导致前列腺组织中雄激素受体表达异常,影响前列腺功能的正常运转,这可能是男性不育或生育能力低下的原因之一。