抗精子抗体阳性大鼠精子凋亡及机制的研究

目的探讨抗精子抗体（AsAb）阳性人鼠精子凋亡率的变化及其可能的机制。方法建立AsAb阳性人鼠动物模型，HE染色观察人鼠附睾组织形态；采用3’-原位末端DNA标记（TUNEL）技术检测大鼠精子的凋亡率；应用化学比色法测定精了匀浆的MDA含量和总抗氧化能力；应用定磷法测定精予匀浆的Na^＋-K^＋-ATP酶和Ca24ATP酶活性。结果与正常组相比，AsAb阳性组大鼠精子凋亡率和MDA含量显著增加（P〈0．01：P〈0．01）；总抗氧化能力、Na^＋-K^＋-ATP酶和Ca^2＋-ATP酶活性显著降低（P〈0．01；P〈0．01；P〈0．05）。结论AsAb阳性大鼠精子凋亡率增加；可能与氧自由基含量增多、Na^＋K^＋-ATP酶和Ca^2＋-ATP酶活性降低有关。     

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。     

青春期对隐睾术后抗精子抗体的影响

目的:检测青春期前及青春期隐睾术后者血清抗精子抗体(antisperm antibodies,AsAb),探讨青春期对隐睾术后血清AsAb的影响。方法:抽取青春期前隐睾术后者84人及青春期隐睾术后者92人静脉血,采用抗精子抗体免疫斑点法检测检测AsAb Ig-G,Ig-M水平。结果:两组AsAb的阳性率差异有统计学意义(P<0.05)。结论:隐睾术后进入青春期者AsAb在血清中阳性率较隐睾术后未进入青春期者显著增高。     

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。     

睾丸活检前后血清抗精子抗体检测

睾丸活检为男性不育症诊断方法之一。属于损伤性检查方法。为确定睾丸活检是否破坏血睾屏障产生自身抗精子抗体。我院对 4 0例不育症患者进行睾丸活检前后血清抗精子抗体检测分析。资料与方法一、一般资料本组年龄在 2 0～ 36岁 ,平均为 2 8岁。均已结婚 ,婚期均在 2年以上 ,睾丸重量约在 5～ 35ｇ ,睾丸长度约在 1.5～ 3.6ｃｍ ,宽度约在 1.2～ 3.3ｃｍ ,厚度约在 1.0～ 1.8ｃｍ。患者性功能基本正常。二、方法1.睾丸活检　常规消毒皮肤后 ,用 2 %利多卡因局麻下作 1～ 2ｃｍ阴囊切口 ,逐层分开阴囊各层 ,切开白膜 ,用尖锐剪刀将睾丸组织切…     

血清抗精子抗体阳性对青春期雄性大鼠睾丸组织及睾丸生殖细胞中Fas/Fas-L凋亡途径的影响

目的:通过人工免疫雄性大鼠获得抗精子抗体(AsAb)介导的血清AsAb阳性的免疫性不育大鼠动物模型,观察AsAb对青春期大鼠睾丸组织及睾丸生殖细胞中Fas/Fas-L凋亡途径的影响。方法:5周龄(青春期)雄性Wistar大鼠30只,其中10只处死,取精子制备精子悬液免疫大鼠,余20只动物随机分为实验组(10只)和对照组(10只),4周后摘取睾丸。光镜观察睾丸组织改变,免疫组化法检测Fas、Fas-L和Caspase-3蛋白的表达。结果:实验组睾丸组织切片呈凋亡样改变,实验组Fas、Fas-L及Caspase-3蛋白的OD值(176.97±4.58,187.52±7.76,157.65±7.38)较对照组(161.87±5.37,150.27±8.65,120.37±6.76)显著增高(P<0.01)。结论:同种精子免疫大鼠,可成功制作AsAb介导的免疫性不育模型;AsAb影响雄性大鼠的生育力,其机制可能与Fas/Fas-L凋亡途径中Fas、Fas-L和Caspase-3蛋白的表达升高有关。     

人类抗精子抗体对精子功能的影响：1.人类抗精子抗体对精子穿越宫颈…

为探讨抗精子抗体对精子穿越宫颈粘液能力的影响，本文用１５份含高滴度抗精子抗体的血清配成不同稀释度的血清标本，用间接精子免疫珠结合试验测定各血清标本的抗体滴度，并用精子宫颈粘液穿透试验检测不同滴度抗精子抗体对精子穿越宫颈粘液能力的影响。     

不同睾丸活检术前后血清抗精子抗体的检测

免疫性不育在男性不育中约占10％～30％，抗精子抗体（antisperm antibody，AsAb）在免疫性不育中起着关键作用，它可通过多种环节干扰生殖…。睾丸活检曾经是唯一对无精子症或少精子症有诊断价值的检查，20世纪90年代以来，由于卵细胞胞质内单精子注射（ICSI）的广泛开展，睾丸活检不仅是一种诊断方法，亦成了一种治疗手段，但其术后可能产生AsAb。1999年1月～2004年10月我们对不同方法睾丸活检术前后AsAb进行了检测，现报告如下。     

Effect of NO on spontaneous acrosome reaction in AsAb positive rat spermatozoa

Objective: To explore the effect of NO on the spontaneous acrosome reaction in antisperm antibody (AsAb) positive rat spermatozoa. Methods: The rat model of AsAb was set up by artificial immunization. The level of AsAb in blood serum was determined by TAT and ELISA. Rat spermatozoa was visualized by staining the acrosome with Coomassie brilliant blue. The NO concentration in rat spermatozoa was assayed by HPLC. Results: The percentage of acrosome reaction, NO concentration, superoxide dismutase (SOD) and Na~ -K~ ATPase activity in AsAb positive rat spermatozoa were significantly decreased compared with the control group. Low dose of NO (SNP 10~(-9)～10~(-8) mol/L) increased the percentage of acrosome reaction and SOD activity, but had no effect on Na~ -K~ ATPase activity. High dose of NO (SNP 10~(-6)～10~(-4) mol/L) decreased the three items. Conclusion: The decrease in acrosome reaction in positive AsAb rat spermatozoa might be related to a decrease in NO and increase in O_2. (the SOD activity was decr     

Impact of silymarin on cadmium-induced apoptosis in human spermatozoa

Oxidative stress-induced apoptosis in spermatozoa may lead to male infertility. Environmental pollutants and heavy metals such as cadmium cause harmful effects on the reproductive system and sperm parameters through the induction of oxidative stress. Silymarin, as a potent antioxidant, is able to inhibit oxidative stress. This study was performed to investigate the protective effects of silymarin on cadmium-induced toxicity in human spermatozoa. Sperm samples were divided into the following five groups: (a) spermatozoa at 0 min, (b) spermatozoa in the control group, (c) spermatozoa treated with cadmium chloride (20 μM), (d) spermatozoa treated with silymarin (2 μM)+ cadmium chloride (20 μM) and (e) spermatozoa treated with silymarin (2 μM). Sperm parameters related to apoptosis, such as DNA fragmentation, nucleus diameter, mitochondrial membrane potential (MMP) and expression of caspase-3, were evaluated in all groups. After 180 min, spermatozoa treated with cadmium chloride showed a significant decrease in nucleus diameter and MMP but a significant increase in DNA fragmentation; however, caspase-3 expression remained unchanged. At this time point, silymarin in the silymarin + cadmium chloride group could significantly reverse the adverse effects of cadmium chloride on these parameters.Silymarn could partly compensate for the caspase-independent apoptosis in the spermatozoa. Therefore, oxidative stress could be a consequence for cadmium toxicity.     

过敏康Ⅱ号胶囊对AsAb阳性大鼠睾丸Bcl-2、Bax表达影响的实验研究

目的观察过敏康Ⅱ号胶囊对AsAb阳性大鼠睾丸Bcl-2、Bax表达的影响。方法选取健康成年雄性SD大鼠60只，按体重随机分为正常组，模型组，对照组，高、中、低剂量组，每组10只。采用主动免疫法建立血清抗精子抗体（AsAb）阳性动物模型10只，灌胃给药，免疫组化方法观察药物对AsAb阳性大鼠睾丸Bcl-2、Bax表达的影响。结果过敏康Ⅱ号高剂量组睾丸生精细胞和精子Bcl-2表达的平均吸光值显著高于模型组（P〈0．01），而Bax表达的平均吸光值显著低于模型组（P〈0．01）。结论调节睾丸Bcl-2、Bax的表达是过敏康Ⅱ号清除或抑制AsAb起治疗作用的机制之一。     

人参皂甙对去势大鼠阴茎海绵体组织NO含量与细胞凋亡的影响

目的研究人参皂甙对去势大鼠阴茎海绵体组织细胞凋亡、NO含量的影响,探讨人参皂甙壮阳功效的可能机制。方法40只成年雄性大鼠随机分为去势组、对照组及不同剂量(25mg/kg、100mg/kg)人参皂甙组共4组,1周后取阴茎海绵体,放免法检测血清睾酮含量(ng/ml),全自动生化分析仪比色法测定海绵体NO含量(μg/g),末端脱氧核糖核酸转移酶介导的duTP缺口末端标记法测定细胞凋亡。结果对照组血清睾酮水平浓度为(1.51±0.86),在去势组、人参皂甙治疗组(25mg/kg、100/mg/kg)均未测到。去势组阴茎海绵体NO浓度(14.45±2.38)较对照组(39.8±3.28)显著降低(P<0.01),25mg/kg人参皂甙组阴茎海绵体NO水平(16.02±2.67)与去势组(14.45±2.38)接近(P>0.05),100mg/kg人参皂甙组NO水平(37.88±7.06)较去势组细胞凋亡数(14.45±2.38)明显升高(P<0.05),与对照组(39.8±3.28)接近(P>0.05)。大剂量100mg/kg人参皂甙组(12.51±1.81)较去势组(26.02±5.25)低(P<0.05)。25mg/kg人参皂甙组凋亡细胞积分光密度(27269.60±4920.42)与去势组比较(33931.50±2459.36)差异无统计学意义,大剂量100mg/kg人参皂甙组(18766.36±3040.42)较去势组(33931.54±2459.36)低,两者比较差异有统计学意义(P<0.05)。结论100mg/kg剂量的人参皂甙不能增加去势大鼠血清睾酮含量,但可以提高去势大鼠阴茎海绵体组织NO水平,减少海绵体细胞凋亡。人参皂甙对去势大鼠阴茎海绵体细胞凋亡的抑制作用可能与其增加NO水平有关。     

缺血预处理对大鼠急性缺血-再灌注肾细胞凋亡的影响

目的:探讨缺血预处理对急性肾缺血-再灌注细胞凋亡的影响。方法:采用8 min缺血加5 min再灌注预处理在体肾缺血-再灌注模型(I45 min I-R 6h),将实验动物随机分为正常(A组)、假手术(S组)、单纯缺血(B组)、缺血再灌注(C组)、预处理(D组)五组,采用透射电镜、流式细胞仪检测肾细胞凋亡和细胞增殖周期,利用光学显微镜进行组织学观察,同时测定血清中尿素氮(BUN)、肌酐(Cr)及MDA含量。结果:与A、S组相比,C组细胞凋亡率显著增高(P<0.01);与C组相比,D组细胞凋亡率和细胞增殖指数均降低(P<0.01),G0/G1时段增加(P<0.01),肾组织损伤病理评分显著降低(P<0.05),同时肾超微结构破坏较轻。结论:缺血预处理对急性肾缺血-再灌注有保护作用,可以减轻急性肾缺血-再灌注引起的细胞凋亡,其作用机制可能与调节细胞增殖周期有关。     

Effect of local testicular heating on spermatogenic cell apoptosis in rats

Aim: To study the effect of testicular local heating on spermatoge-nic cell apoptosis in rats. Methods: Seventy male SD rats were divided into the heat-treated and the control groups. The former was exposed to heat (43 ℃) for 12 hours. Each group was further divided into seven subgroups with respect to the time of observation after heat exposure, i.e., 12 h and 1 days, 3 days, 6 days, 10 days, 50 days and 80 days, respectively. In each subgroup, sper-matogenic cell apoptosis was examined by means of electron microscopy, flow cytometry and terminal deoxynucleotidyl trans-ferase-mediated dUDP-nick end labeling (TUNEL) methods. Results: The percentage of cells with sub-haploid and the percentage of positive TUNEL cells were significantly higher in the heat-treated groups than in the controls (P<0.01). The reaction of cell apoptosis to local heat was highly selective: spermatocytes were the most sensitive, followed by spermatids, spermatozoa and sper-matogonia in a decreasing order. Conclusion: Local testic     

七氟醚预先给药对大鼠肾脏缺血再灌注时细胞凋亡的影响

目的 探讨七氟醚预先给药对大鼠肾脏缺血再灌注时细胞凋亡的影响.方法 健康清洁级雄性SD大鼠30只,体重220～260 g,采用随机数字表法,将大鼠随机分为3组(n=10):对照组(C组)、缺血再灌注组(I/R组)、七氟醚组(S组).I/R组和S组采用夹闭左肾蒂45 min后恢复再灌注的方法 建立肾脏缺血再灌注模型,C组腹部正中切口,右肾切除,左肾蒂游离后,缝合腹腔;S组模型制备前30 min开始吸入2.2%七氟醚和氧气的混合气体至再灌注3 h.于再灌注3 h时采集下腔静脉血样5 ml,测定血清尿素氮(BUN)、肌酐(Cr)浓度,然后取肾组织,光镜下观察肾组织病理学结果,TUNEL法检测细胞凋亡,计算细胞凋亡指数,采用RT-PCR和Western blot法测定血红素氧合酶-1(HO-1)mRNA及蛋白表达水平.结果 与C组比较,I/R组和S组血清BUN、Cr浓度、肾脏近曲小管坏死程度、细胞凋亡指数升高,HO-1 mRNA和蛋白表达上调(P＜0.05);与I/R组比较,S组血清BUN、Cr浓度、细胞凋亡指数、肾脏近曲小管坏死程度降低,HO-1 mRNA表达上调(P＜0.05).结论 七氟醚预先给药可通过抑制细胞凋亡而减轻大鼠肾脏缺血再灌注损伤,其抑制细胞凋亡作用可能与HO-1 mRNA表达上调有关.

Abstract：

Objective To investigate the effects of sevoflurane pretreatment on renal ischemia-reperfusion (I/R)-induced apoptosis in kidney in rats. Methods Thirty pathogen-free male SD rats weighing 220-260 g were randomized into 3 groups (n=10 each):group control (group C);group I/R and group sevoflurane(group S). Renal I/R was induced by clamping the left renal pedicle for 45 min in I/R and S groups. In group S inhalation of 2.2% sevoflurane in O2 was started at 30 min before operation and maintained throughout the experiment.Venous blood samples were taken at 3 h of reperfusion for determination of serum BUN and Cr concentrations. The animals were then sacrificed and the left kidneys were removed for microscopic examination, detection of apoptosis(by TUNEL)and determination of heme oxygenase-1(HO-1) mRNA and protein expression (by RT-PCR and Western blot).Results Renal I/R significantly increased serum BUN and Cr concentrations, apoptotic index(percentage of apoptotic cells) and the severity of necrosis of renal proximal convoluted tubules (0=normal,4=necrosis of whole segment of proximal convoluted tubules).Sevoflurane inhalation attenuated the I/R-induced changes mentioned above.HO-1 mRNA and protein expression was up-regulated by I/R and HO-1 mRNA expression was further up-regulated by sevoflurane inhalation.Conclusion Sevoflurane pretreatment can protect kidney against I/R injury by attenuating cell apoptosis.Up-regulation of HO-1 mRNA expression may be involved in the mechanism.     

肿瘤坏死因子相关的凋亡诱导配体基因在糖尿病大鼠肾脏细胞凋亡中的作用

目的 探讨肿瘤坏死因子(TNF)相关的凋亡诱导配体(TRAIL)基因在糖尿病大鼠肾脏细胞凋亡中的表达及其作用。方法 应用链脲佐菌素建立糖尿病大鼠模型，采用原位末端标记法和流式细胞术检测4组(对照组、糖尿病4周组、8周组和12周组)大鼠肾脏细胞凋亡情况；免疫组化和流式细胞术检测肾脏TRAIL基因及其死亡受体DR4、DR5的蛋白表达。结果 与正常对照组相比．各糖尿病组较对照组肾小球、肾小管凋亡细胞数明显增多，TRAIL、DR4、DR5的表达亦显著增强。结论 在高糖环境的诱导下．TRAIL基因及其死亡受体出现的高表达，可能部分参与了糖尿病大鼠肾脏细胞凋亡。     

亚麻木酚素对大鼠生长板软骨细胞凋亡的影响

目的：比较不同浓度亚麻木酚素抑制生长板软骨细胞凋亡的效果,筛选出最适药物浓度,为其延迟大鼠骨骺闭合促进长骨生长进行理论支持。方法：选取2只4周龄雄性SD大鼠,SPF级,体质量80 g。解剖大鼠胫骨及股骨生长板软骨并进行体外分离,获得生长板软骨细胞进行培养,利用倒置相差显微镜及Ⅱ型胶原免疫荧光实验对软骨细胞分别进行形态学观察、鉴定,然后采用白细胞介素-1β(interleukin-1β,IL-1β)20 ng/ml诱发生长板软骨细胞凋亡为模型组,同时加入分别含1、10、20、40μM亚麻木酚素为实验组,同时加入5μM来曲唑为阳性对照组,分别培养24、48 h,MTT法观察药物促进细胞增殖情况,流式细胞仪检测药物抑制细胞凋亡情况。结果：含量1、10、20、40μM的亚麻木酚素均可不同程度促进细胞增殖,其原理为药物抑制了IL-1β诱发的生长板软骨细胞凋亡,抑制细胞凋亡的最佳药物浓度为20μM。结论：适宜浓度的亚麻木酚素对大鼠生长板软骨细胞凋亡抑制效果明显且最适药物浓度为20μM,为骨骼延迟闭合提供更长的生长时间,从而为促进发育期骨骼增长提供可能。