皮质酮诱导大鼠Leydig细胞凋亡是一经caspase-3激活的过程

目的 超生理剂量的皮质酮(大鼠体内的糖皮质激素)能诱导大鼠Leydig细胞凋亡。但有关皮质酮诱导Leydig细胞凋亡的细胞内机制尚不清楚。本研究旨在观察皮质酮是否经caspase-3激活的途径诱导大鼠Leydig细胞凋亡。方法 采用Western Blot方法检测不同时间点上经皮质酮处理的大鼠Leydig细胞中caspase-3酶原及裂解的caspase-3酶表达情况。运用荧光发光法检测不同时间点上经皮质酮处理的人鼠Leydig细胞中caspase-3酶活性。结果 caspase-3酶原表达水平在皮质酮处理6h时开始上升，12h及24h时表达量下降，而具生物活性的、裂解的caspase-3酶于12h时开始出现，24h时的表达水平最为显著。caspase-3酶活性在皮质酮处理12h时明显升高，以24h时最为显著。Caspase-3抑制剂DEVD-CHO对经皮质酮处理12、24及48h的Leydig细胞中的caspase-3酶活性均具有明显的抑制作用，加caspase-3抑制剂的处理组其细胞基因组DNA电泳未见有凋亡特征性的梯状条带。结论 皮质酮诱导的大鼠Leydig细胞凋亡是一经caspase-3激活的过程。     

皮质酮诱导的Leydig细胞凋亡中Egr的表达及其作用

目的评价皮质酮是否通过活化钙调神经磷酸酶（CaN）来诱导Leydig细胞中转录因子Egr-2及Egr-3的表达及两种Egr对皮质酮诱导的Leydig细胞凋亡的影响。方法采用RT-PCR方法检测CaN的抑制剂环孢菌素A（CsA）对经皮质酮处理的Leydig细胞中Egr-2及Egr-3的mRNA表达水平；用Annexin-V-FITC和PI双标法评价Egr-2和Egr-3的表达载体对Leydig细胞凋亡率的影响。结果皮质酮能诱导Leydig细胞中Egr-2及Egr-3的表达，且该诱导作用可被CsA抑制；过量表达的Egr-2及Egr-3均可使经皮质酮诱导的Leydig细胞凋亡率增加，并以Egr-3的作用较为显著。结论高浓度的皮质酮可通过活化CaN诱导Leydig细胞中Egr．2和Egr．3的表达，两种Egr对经皮质酮诱导的Leydig细胞的凋亡均具有促进作用。     

Egr参与调控皮质酮处理的睾丸Leydig细胞中FasL启动子的活性

目的观察转录因子Egr(early growth response factor,早期生长反应因子)是否参与调控皮质酮处理的Leydig细胞中FasL启动子的活性。方法经RT-PCR检测皮质酮处理的Leydig细胞中Egr-2及Egr-3的mRNA的水平,采用双荧光素酶报告基因系统评价Egr-2和Egr-3表达载体对Leydig细胞中FasL启动子活性的影响。结果Leydig细胞经皮质酮处理后,细胞内Egr-2及Egr-3在转录水平呈显著上调。双荧光素酶报告基因检测发现,Egr-2及Egr-3表达载体均可上调皮质酮处理的Leydig细胞中FasL启动子的活性,并以Egr-3的作用为显著。结论Egr-2及Egr-3参与调控皮质酮处理的Leydig细胞中FasL启动子的活性。     

皮质酮诱导的大鼠Leydig细胞凋亡中caspase-3活化途径的研究

目的我们新近的研究表明，超生理剂量的皮质酮（大鼠体内的糖皮质激素）可通过激活caspase-3诱导大鼠Leydig细胞凋亡。本研究拟评价在皮质酮诱导的大鼠Leydig细胞凋亡过程中，caspase-3的激活是否有其上游的caspase-8平和 caspase-9的参与。方法采用荧光分光光度法榆测经皮质酮处理的大鼠Leydig细胞中caspase-8活性，以DNA梯状电泳条带作为评价细胞凋亡的指标，观察caspase-8抑制剂是否能够抑制细胞凋亡，采用RTPCR枪测皮质酮诱导的大鼠Leydig细胞中caspase-9的mRNA水平。结果在皮质酮诱导的大鼠Leydig细胞中出现caspase-8活性增高，以12h最为址著，升高的caspase-8的活性可被caspase-8抑制剂抑制，并导致Leydig细胞的凋亡过程被阻断。Leydig细胞中caspase-9的mRNA水平在皮质酮作用下上升，同样以12h最为显著。结论皮质酮诱导的大鼠Leydig细胞调亡与caspase-8和caspase-9有关。     

Egr在高浓度皮质酮诱导的大鼠Leydig细胞凋亡中的表达及作用

目的 观察在高浓度皮质酮处理的大鼠Leydig细胞中Egr mRNA的表达量,并评价Egr在高浓度皮质酮诱导的Leydig细胞凋亡的作用.方法 采用RT-PCR方法检测经高浓度皮质酮以及高浓度皮质酮加环孢菌素A (CsA)处理的Leydig细胞中早期生长反应因子(early growth response factor,Egr) Egr-2及Egr-3的mRNA表达水平:用Annexin-V -FITC和Pl双标法检测Egr-2和Egr-3表达载体对经高浓度皮质酮诱导的Leydig细胞凋亡率的影响.结果 经高浓度皮质酮处理的Leydig细胞其Egr-2及Egr-3mRNA的表达量均显著升高,而皮质酮的这一作用能被钙调神经磷酸酶(CaN)的抑制剂环孢菌素A(CsA)所抑制.Leydig细胞中过量表达的Egr-2及Egr-3均可促进皮质酮诱导细胞凋亡,并以Egr-3的作用较为显著.结论 高浓度的皮质酮可能通过活化CaN来诱导Leydig细胞中Egr-2和Egr-3的表达,由此促进了Leydig细胞的凋亡过程.     

敌敌畏对大鼠睾丸Leydig细胞凋亡的影响

目的:观察敌敌畏对子代雄性大鼠睾丸Leydig细胞凋亡的影响,探讨Leydig细胞变化在泌尿生殖系统畸形发病机制中的意义。方法:将21只孕SD大鼠随机分为对照组和敌敌畏各剂量组,在孕12～17 d期间,对照组每天分别给予灌喂玉米油1.0 ml;敌敌畏各剂量组每天分别给予敌敌畏1、4、8、16、20、24 mg/kg。待孕鼠分娩完毕,每组随机选取5只新生雄性仔鼠睾丸标本行HE染色、caspase-3免疫组化染色和DAPI荧光染色,将余下的雄性仔鼠饲养至90 d后再随机选取5只成年雄性仔鼠睾丸标本行HE染色、caspase-3免疫组化染色和DAPI荧光染色。结果:玉米油对照组雄性仔鼠睾丸组织caspase-3染色阳性的Leydig细胞数量和DAPI标记的Leydig细胞的总分值与敌敌畏1 mg/(kg.d)组比较无统计学意义(P>0.05),与敌敌畏其余剂量组比较有统计学差异(P<0.05),敌敌畏可使雄性仔鼠睾丸caspase-3染色阳性的Leydig细胞数量和DAPI标记的Leydig细胞的总分值增加,并与敌敌畏存在剂量依赖关系。结论:孕期染毒敌敌畏可使雄性仔鼠睾丸Leydig细胞的凋亡增加,这可能影响到Leydig细胞的数量,使胚胎期胎睾产生睾酮的功能受到干扰,从而使泌尿生殖系统的发育受到影响。     

皮质酮诱导青春期大鼠睾丸间质细胞凋亡的研究

本研究目的旨在观察皮质酮能否诱导青春期大鼠睾丸间质细胞凋亡,用皮质酮分别经体内、外途径处理大鼠睾丸间质细胞。凋亡细胞经碘化丙碇（ＰＩ）标记后用流式细胞仪检测。体外研究结果表明,经１００ｎＭ皮质酮处理２４小时后的睾丸间质细胞,其凋亡量（３６．５％）显著高于对照组（１２．７％。Ｐ〈０．０１）。在体研究得到类似结果,经皮质酮（２．５ｍｇ／１００ｇ体重）处理２４小时后的大鼠,其纯化的睾丸间质细胞调亡量（２     

瘢痕细胞凋亡过程中Ca2+和三磷酸肌醇浓度的改变

目的：探讨顺铂诱导瘢痕细胞凋亡过程中细胞内游离Ca^2 浓度和三磷酸肌醇(IP3)浓度的变化特点和意义。方法：选择顺铂诱导瘢痕细胞凋亡，采用光镜、电镜、琼脂糖凝胶电泳和流式细胞术检测凋亡。Fura-2荧光负荷技术测(Ca^2 )i，竞争性蛋白结合法测定IP3浓度。结果：顺铂浓度为2ug／mL作用于瘢痕成纤维细胞24小时出现典型的凋亡改变。瘢痕细胞凋亡过程中，细胞内(Ca^2 )i明显升高，以凋亡早期更为明显。瘢痕细胞凋亡早期，IP，浓度明显升高，晚期IP3浓度明显下降。结论：在瘢痕细胞凋亡过程中，(Ca^2 )i表现为上调趋势。IP3浓度在凋亡早期，表现为上调趋势，在稍晚阶段，表现为下调趋势，这为治疗瘢痕提供了新靶点。     

Cox7a2和ATP50在原代培养小鼠Leydig细胞和人体不同器官的表达研究

目的 研究线粒体呼吸链相关基因Cox7a2,ATP50在原代培养的小鼠睾丸Leydig细胞和人体各器官组织中的表达特征.方法 原代培养小鼠睾丸Leydig细胞并鉴定,利用RT-PCR方法 研究Cox7a2,ATP50在Leydig细胞中的表达.利用PCR方法 研究Leydig在人体各重要脏器组织的表达谱.结果 Cox7a2,ATP50在原代小鼠睾丸Leydig细胞中表达,Cox7a2和ATP50在各个脏器旱现出不同的表达特征.结论 Cox7a2,ATP50表达在原代小鼠睾丸Leydig细胞中,研究Cox7a2,ATP50在人体各器官的表达特点,为基因的功能研究奠定了前期基础.     

衰老的Leydig细胞中睾酮合成降低的机制

睾酮是男性体内主要的雄激素。老年男性体内睾酮水平下降使机体呈现肌肉松弛、骨质疏松和性欲减退等衰老现象，这与睾酮合成减少及其分泌节律改变密切相关。除了。肾上腺皮质有少量分泌以外，睾酮主要由睾丸间质中的Leydig细胞生成。对于衰老Leydig细胞中睾酮合成减少的机制的研究已有不少报道，影响     

Involvement of nuclear factor-kappa B on corticosterone- induced rat Leydig cell apoptosis

Aim:To investigate the activation of nuclear factor-kappa B(NF-kappa B)and its function in glucocorticoid-inducedLeydig cell apoptosis.Methods:The Leydig cells were isolated from male Sprague-Dawley rats(90 days of age)andwere incubated with corticosterone(CORT,glucocorticoid in rat)for 6 h,12 h and 24 h,respectively.The P65subunit of NF-kappa B(NF-kappa B/P65)in nuclei and the inhibitor of NF-kappa B(Ikappa B)in cytoplasm wereanalyzed by Western-blotting.The Leydig cells were treated with anti-Fas antibody for 3 h followed by Westernblotting to assay the changes of NF-kappa B/P65 in nuclei and in cytoplasm.The role of NF-kappa B in CORT-induced Leydig cell apoptosis was evaluated by observing the effects of NF-kappa B/P65 overexpression and inhibit-ing activation of NF-kappa B by 100μmol/L Pyrrolidine dithiocarbamate(PDTC)on this apoptosis.Results:Thetreatment of Leydig cells with CORT increased the levels of NF-kappa B/P65 in nuclei and decreased the levels ofIkappa B in cytoplasm.Following the Leydig cells were treated with anti-Fas antibody,the levels of NF-kappaB/P65was increased in nuclei and decreased in cytoplasm.The CORT-induced Leydig cell apoptosis was inhibited byoverexpressed NF-kappaB/P65 and was enhanced by incubation with PDTC.Conclusion:NF-kappa B is activatedby increased FasL/Fas in CORT-induced Leydig cell apoptosis.NF-kappa B may play an anti-apoptotic role in thisapoptosis.(Asian J Androl 2006 Nov;8:693-702)     

NFAT2 is implicated in corticosterone-induced rat Leydig cell apoptosis

Aim： To investigate the activation of the nuclear factor of activated T cells （NFAT） and its function in the corticosterone （CORT）-induced apoptosis of rat Leydig cells. Methods： NFAT in rat Leydig cells was detected by Western blotting and immunohistochemical staining. Cyclosporin A （CsA） was used to evaluate potential involvement of NFAT in the CORT-induced apoptosis of Leydig cells. Intracellular Ca^2＋ was monitored in CORT-treated Leydig cells using Fluo-3/AM. After the Leydig cells were incubated with either CORT or CORT plus CsA for 12 h, the levels of NFAT2 in the nuclei and in the cytoplasm were measured by semi-quantitative Western blotting. The role of NFAT2 in CORT- induced Leydig cell apoptosis was further evaluated by observing the effects of NFAT2 overexpression and the inhibition of NFAT2 activation by CsA on FasL expression and apoptosis. Results： We found that NFAT2 was the predominant isoform in Leydig cells. CsA blocked the CORT-induced apoptosis of the Leydig cells. The intracellular Ca^2＋ level in the Leydig cells was significantly increased after the CORT treatment. The CORT increased the level of NFAT2 in the nuclei and decreased its level in the cytoplasm. CsA blocked the CORT-induced nuclear translocation of NFAT2 in the Leydig cells. Both CORT-induced apoptosis and FasL expression in the rat Leydig cells were enhanced by the overexpression of NFAT2 and antagonized by CsA. Conclusion： NFAT2 was activated in CORT-induced Leydig cell apoptosis. The effects of NFAT2 overexpression and the inhibition of NFAT2 activation suggest that NFAT2 may potentially play a pro-apoptotic role in CORT-induced Leydig cell apoptosis through the up-regulation of FasL.     

睾丸间质细胞瘤2例报告并文献复习

目的 探讨睾丸间质细胞瘤的临床病理特点,进而提高睾丸间质细胞瘤的诊治水平。方法 回顾性分析2例睾丸间质细胞瘤的临床病理资料并复习相关文献。结果 1例术中快速冷冻切片病理诊断为睾丸间质细胞瘤,行根治性睾丸切除术,1例直接行根治性睾丸切除术,术后病理均诊断为睾丸间质细胞瘤,其中1例精索残端及周围脂肪组织可见肿瘤细胞,精索静脉可见肿瘤浸润。术后分别随访24和6个月,未见复发或转移。结论 睾丸间质细胞瘤临床罕见,术前诊断较为困难,确诊仍依赖组织病理学检查,外科手术是睾丸间质细胞瘤唯一有效的治疗方法。     

Epididymal leiomyoma associated with leydig cell hyperplasia: A case report and immunohistochemical study

Leiomyomas are rare soft tissue neoplasms thatcan affect virtually any part of themale genital tract. Although they have beendescribed in association with other lesions, tobest our knowledge coexistence with leydig cellhyperplasia has not been reported before.We present here the clinical, histopathologicand immunhistochemical characteristics of anepididymal leiomyoma associated with leydigcell hyperplasia and an epididymal cyst in asixty-eight year old man.     

Effects of varicocele on testosterone, apoptosis and expression of StAR mRNA in rat Leydig cells

The aim of this study was to explore the effects of varicocele on the morphology and function of Leydig cells in the rat testis. Forty male Sprague-Dawley rats were divided into two groups: the experimental group underwent surgery to create a left varicocele (VC), and the control group underwent a sham operation. Serum testosterone and intratesticular testosterone levels were measured using a radioimmunoassay after 4 and 8 weeks of operation. Leydig cells were studied for apoptosis and expression of steroidogenetic acute regulatory (StAR) protein mRNA levels. Serum testosterone levels declined after 4 and 8 weeks of operation but were not significant (P>0.05). However, the intratesticular testosterone levels after 8 weeks were significantly decreased compared with the control group (P<0.01). The mean apoptosis index of Leydig cells in the experimental group was significantly higher than that in the control group after 4 or 8 weeks (P<0.01). StAR mRNA levels in the Leydig cells of the experimental group were significantly lower compared to those of the control group (P<0.01). Our data show that varicocele did impair Leydig cell function by increasing apoptosis and suppressing the expression of the StAR protein.     

补骨颗粒含药血清对大鼠软骨细胞凋亡及Trx2信号通路的影响

目的观察补骨颗粒含药血清对体外培养的大鼠膝关节软骨细胞凋亡及对Trx2、ASK1及Caspase3表达的影响,从而探讨补骨颗粒预防骨性关节炎发生发展的作用机制。方法采用两步酶消化法分离培养大鼠软骨细胞,并进行传代培养。应用膜联蛋白V-FITC/PI染色后经流式细胞仪检测软骨细胞的凋亡情况。同时,通过电泳分离蛋白并通过蛋白质印迹分析Trx2、ASK1及Caspase3的表达情况。结果软骨细胞培养15 d左右铺满80%～90%的培养皿,大部分细胞呈梭形。流式细胞检测结果显示空白血清组的细胞凋亡率为22.80%,明显高于含药血清组(P0.05),而20%含药血清组(15.91%)与30%含药血清组(17.93%)的细胞凋亡率又明显低于10%含药血清组(21.58%),各组间比较差异具有统计学意义(P0.05)。蛋白质印迹分析结果显示含药血清组Trx2的表达量都明显增多,以10%含药血清组的表达量最多;空白组与10%含药血清组的ASK1与Caspase3的表达量比20%与30%含药血清组多。结论补骨颗粒可以通过激活Trx2信号通路而抑制软骨细胞的凋亡,从而起到预防骨性关节炎发生发展的作用。