棉酚对人精子乳酸脱氢酶-X 的影响

对10例服棉酚者的精子作LDH同功酶组织化学检查和电泳观察,结果:①精子颈体部的组化反应形成的沉淀主要是LDH-X,服药后沉淀变粗,数量减少以至消失;LDH-X占总LDH的平均含量百分率为42.6％,服药30、50和100天后则分别为31.1,26.3和10.2％,下降显著,提示棉酚可干扰精子的能量代谢。②LDH-X的变化程度因个体对棉酚的敏感性不同而有差异。LDH-X组化检查法简便、特异,可作为判断精子质量的敏感指标。     

醋酸棉酚与磺胺嘧啶对人精子LDH-X酶及活力的协同作用研究

棉酚抗生育作用业已被国内外学者所证实(1～4)。并对其作用部位、机理、代谢、毒性等进行了深入地研究[5]。分子及酶水平的分析表明,棉酚可抑制三羧酸循环中数种酶的活性[6,7]。Lee等[8]认为棉酚是LDH-X的一种选择性抑制剂。另一方面,有学者[9]证明磺胺类药物亦具有杀伤精子的作用。我们初步实验也证实磺胺嘧啶(SD)可抑制人精子LDH-X活性及对精子有直接杀伤作用。本研究旨在观察在体外条件下醋酸棉酚与SD是否具有抑制人精子LDH-X与杀伤精子的协同作用。现将实验结果报道如下。     

人睾丸乳酸脱氢酶同工酶-X的纯化和它的免疫性质

乳酸脱氢酶(LDH)及其同工酶(1～5),广泛存在于哺乳动物各类组织细胞中。1963年Blanco等和Goldberg等首先在人精液中发现了LDH-X。Glement证实LDH-X仅存在于睾丸的初级精母细胞、精子细胞、精子和精液中,并以精子中的含量最高(约占总量的80～90％),在精子中含量高的事实,提示LDH-X的存在可能与精子的能量代谢有关。近年来,注意到抗LDH-X的血清能抑制豚鼠精     

雷公藤单体（T4）与棉酚抗生育机理及毒性作用的比较

喂服雷公藤单体(T_4)0.05mg/kg/d6周后,大鼠生精作用及晚期精子细胞碱性核蛋白转换与替代受到抑制程度均较喂服棉酚组明显。实验组附睾上皮腔缘部γ-GT,ALP出现成簇的阳性沉淀。附睾尾部精子头尾断裂,断端卷曲,尾部纵行粗纤维脱位,线粒体LDH,LDH-X,MDH受到抑制。T_4对肾脏、肝脏各种酶虽有轻度影响但没有引起酶活性明显的变化。T_4对内脏的影响较棉酚轻,但致雄性不育的效果比较明显。本研究提示T_4是一种有发展前途的男性抗生育药物。     

棉酚对成熟公猪精子的呼吸链和氧化磷酸化的解偶联作用

香港中文大学生化部Wung-Wai Tso等文章:已证明实验动物灌喂棉酚后,在附睾尾部产生死精子,睾丸内出现畸形精子。用成熟猪的精子作体外研究,发现棉酚使精子不能向前活动。棉酚浓度低时,精子呼吸增强;浓度高时出现抑制现象。虽然已提示这种抑制活动     

醋酸棉酚对人射出精子Ca^2＋内流和大鼠输精管收缩反应的影响

本实验旨在观察醋酸棉酚对人射出精子Ｃａ~（２＋）跨膜转运和对离休大鼠输精管收缩作用的影响。结果表明，醋酸棉酚抑制人射出精子Ｃａ~（２＋）内流，并呈量－效依赖关系。醋酸棉酚破坏精子膜内、外Ｃａ~（２＋）浓度梯度的动态平衡，可能是其抑制精子活力的重要机理之一。但醋酸棉酚对高Ｋ＋诱发离体大鼠输精管平滑肌的收缩作用没有影响，可能醋酸棉酚对大鼠精子膜有选择性的作用。     

棉酚对雄性大鼠的抗生育作用

棉酚对雄性大鼠有明显的抗生育作用。大鼠口服棉酚12毫克/公斤/天5周,24毫克/公斤/天5周和12毫克/公斤/天12周,仃药时大鼠附睾和输精管精子全部死亡,仃药后抗生育作用可维持4～6周。上述棉酚剂量对雄大鼠的抗生育作用是可逆的。棉酚的抗生育效果和剂量、疗程有关。幼年大鼠口服棉酚10毫克/公斤/天18周,对其生长发育无明显影响,抗生育剂量棉酚对大鼠无明显毒性。     

药物避孕

臾测卿2棉酚对体外家兔精子顶体酶类的抑制作用/袁玉英…//生殖与避孕一1卯6,16‘l)一40-45 本文作者对棉酚体外抑制10种家兔精子顶体酶活性进行了测定。当醋酸棉酚浓度为12一76拼mol几时,可完全或明显抑制顶体酶(Ac二in)、AZOCOn蛋白酶、芳香基硫酸醋酶和神经氨酸昔酶活性;而当浓度高达380拜mol/L时才能抑制透明质酸酶、月一葡糖昔酸酶和酸性磷酸酶,但不抑制磷酸醋酶c、碱性磷酸酶和汗N-乙酞氨基葡糖昔酶。棉酚对芳香基硫酸醋酶的抑制作用是一种非竞争性抑制作用,Ki为120子助01/L。其抑制作用是可逆的,是剂量依赖关系。由于抑制家兔精…     

人睾丸LDH-X的纯化

乳酸脱氢酶(LDH)及其同工酶,(LDH-X)以不同的比例广泛存在于哺乳动物各组织细胞中。1963年Blanco和Goldberg等在人青春期后睾丸及精液、精子中发现LDH-X。LDH-X只存在于精母细胞、精子细胞和精子中。除睾丸外其它器官不产生这种同工酶。LDH-X是由C-亚基组成的四聚体,这种C一亚基是由仅在睾丸中表达的LdhC(乳酸脱氢酶C)基因编码。生精过程中LDH-X的合成起始于初级精母细胞减数分裂前期的粗线中期并持续于整个生精过程中。我们按Wong C法加以改进提纯了人睾丸的LDH-X。材料与方法一、材料及试剂新鲜人睾丸去净包膜、血管及血凝块置     

应用精子头部银染观察棉酚的遗传效应

棉酚对人类的抗生育效果达99％,精子细胞和粗线初级精母细胞对棉酚最为敏感。棉酚可使大鼠精子尾部损伤,出现局部纤维崩解和尾巴弯曲。已知精子头部形态由多基因控制,诱变剂可引起精子头部形态异常,分析精子头部形态改变已成为检验药物对生精细胞遗传效应的敏感指标。Amés试验及对体内、外淋巴细胞染色体影响的研究表明,棉酚既不是诱变剂、致畸剂,亦非致癌物。本文用银染法观察喂服棉酚一年半小鼠及志愿服棉酚者的精子头部形态,以进一步探讨棉酚对生殖细胞有否遗传效应,为其安全使用提供理论依据。     

兔抗人睾丸乳酸脱氢酶同工酶-X抗血清的制备纯化及免疫性质探讨

以纯化的人睾丸乳酸脱氢酶同工酶-X(简称人LDH-X)为抗原免疫家免,制备了兔抗人睾丸乳酸脱氢酶同工酶-X抗血清(简称人LDH-X抗体)。用辛酸法纯化该抗体。以双扩及免疫电泳鉴定,均呈现单一沉淀线。经免疫交叉反应试验证明人LDH-X与人LDH_(1-5)同工酶有不同的免疫决定簇;纯化的人LDH-K抗体只与人睾丸匀浆、人精液呈现单一沉淀线,而与鼠睾丸LDH-X、兔及牛睾丸匀浆均无交叉反应。从而显示纯化的人LDH-X抗体具有较强的种属特异性。     

移动电话电磁辐射对睾丸乳酸脱氢酶同工酶的影响

目的 :探讨移动电话的电磁辐射对雄鼠生殖功能的影响。方法 :用移动电话的模拟辐射源对小鼠进行全身辐射 ,辐射频率为 935MHz ,每天 2h ,连续 35d ,平均功率密度分别为 0 ,5 70 ,14 0 0 μW·cm-2 。照射 35d后取双侧睾丸和附睾称重 ,并观察睾丸乳酸脱氢酶 (LDH)及其同工酶 (LDH X)的活性和睾丸组织结构及电镜超微结构的变化。结果 :各组间的睾丸和附睾的脏器系数差异无显著性 (P >0 .0 5 ) ;但 14 0 0 μW·cm-2 辐射组的LDH X活性显著下降 (P <0 .0 5 ) ,并出现精子超微结构的改变。结论 :移动电话的电磁辐射可以降低睾丸乳酸脱氢酶同工酶活性 ,可能存在生殖毒性。     

着床期大白鼠子宫内膜乳酸脱氢酶同功酶与雌孕激素的关系

用琼脂胶电泳法和光密度计测定大白鼠子宫内膜乳酸脱氢酶同功酶(LDH_(1～5))并计算各同功酶带占LDH_(1～5)总量的百分率。以此分析着床期宫内膜LDH_(1～5)分布形式及雌孕激素对它的影响。实验结果:着床期大白鼠子宫内膜LDH_(1～5)分布以LDH_(4、5)最显著。妊娠第六天后LDH_5显著下降,而LDH_(4、3、2)却明显上升。这种分布形式并非蜕膜组的特点,也不受胚泡植入的影响。我们认为这种分布形式与假孕期及间期的宫内膜的织LDH_(1～5)分布一样,都是在内源性雌激素和孕激素的共同调控下发生的。     

Interaction between aluminum and zinc or copper and its effects on the pituitary-testicular axis. II. Testicular enzyme and serum gonadotropin assay

Sprague-Dawley male rats were divided into groups and maintained on a semipurified diet containing either 5 or 40 ppm of zinc or 2 or 8 ppm of copper. Half of the rats in each group received 1000 ppm aluminum in the diet. It was found that aluminum accumulated in the pituitary glands and testes when dietary copper levels were suboptimal. The ALP activity in testes was depressed by the added aluminum when the intake of zinc or copper was suboptimal. SDH, LDH, and LDH-X activities were inhibited and GRS and GGTP activities were elevated in rats fed either the suboptimal zinc or copper diet. However, the added aluminum in the diet reversed the changes to normal levels. The testosterone levels in plasma changed very little when the zinc or copper intake was suboptimal. An increase in plasma FSH was demonstrated in groups of both suboptimal zinc and copper intake. But the plasma LH was elevated only in the group receiving the suboptimal copper diet, and the added aluminum reversed plasma LH to control levels. A lower level of testosterone was demonstrated in the group given suboptimal copper with aluminum. It was concluded that dietary aluminum influenced the pituitary-testicular axis by interacting with certain essential trace metals, particularly zinc.     

合并应用甾体激素与低剂量棉酚对雄性大鼠的护一育效果 …

目的 论证甾体激素与低剂量棉酚组合用药的雄性抗生育作用的可行性作用机制。方法 用低剂量棉酚１２ｍｇ／ｋｇ（ｋｄ．ｄ）,甲基睾丸素２０ｍｇ（ｋｇ．ｄ）,炔璃００μｇ／（ｋｇ．ｄ）喂服Ｗｉｓｔａｒ大鼠６周,部分取工始合笼;继续用棉酚１２ｍｇ／（ｋｇ．ｄ）维持１２周,部分取材,余下大鼠停药,观察恢复情况。取材做大鼠睾丸精子计数,精子活动度,睾丸切片的形态学观察等。结果 用药６周大鼠的生精过程受到不完全抑     

黄芪甲苷诱导NO生成并激活PKCε对大鼠心肌缺血再灌注损伤的保护作用

目的:研究黄芪甲苷(AsⅣ)对大鼠心肌缺血再灌注损伤的保护作用,阐明其作用机制。方法:30只雄性Wistar大鼠随机分为假手术组、再灌注模型组、阳性药尼可地尔(10mg.kg-1)组、AsⅣ(5mg.kg-1)组及AsⅣ+氯化白屈莱赤碱(CHE)(3mg.kg-1)组,每组6只,结扎左冠状动脉前降支30min,松扎再灌注120min制备大鼠心肌缺血再灌注损伤模型,检测AsⅣ对其血清中LDH、MDA和SOD以及NO的影响,同时光镜下观察心肌细胞病理组织形态学变化,Western blotting法检测心肌细胞胞浆及胞膜PKCε蛋白表达。结果:AsⅣ组与模型组比较,LDH漏出量降低(P<0.05),MDA含量明显降低(P<0.01),SOD活力增强(P<0.01),NO含量增加(P<0.01)。HE染色组织形态学观察,与模型组比较,AsⅣ组心肌细胞形态有明显的改善,炎细胞浸润也有减轻。Western blotting法检测,AsⅣ组心肌细胞胞膜PKCε表达量高于模型组(P<0.05)。CHE减弱了ASⅣ的这种保护作用;与AsⅣ组比较,AsⅣ+CHE组LDH漏出量升高(P<0.05),MDA含量升高(P<0.01),SOD活力降低(P<0.01),PKCε胞膜表达量降低(P<0.05),心肌细胞水肿明显、胞质着色较浅。结论:AsⅣ通过诱导NO的生成对大鼠心肌缺血再灌注损伤产生保护作用,PKCε的激活可能是AsⅣ保护心肌细胞信号传导通路的组成部分。     

Drug synergistic antifertility effect of combined administration of low-dose gossypol with steroid hormones in rats

Background  Our previous studies suggested that low-dose gossypol combined with steroid hormones has a reversible antifertility role in adult male rats, and the course of treatment was shorter than that of either gossypol or steroid hormones alone. This result suggested that low-dose gossypol and steroid hormones have a drug synergistic effect on antifertility. The aim of the study was to find the target organs of the antifertility synergistic effect of the combined regimen.

Methods  Thirty-two adult male rats were divided into four groups randomly: group GH, rats were fed orally with gossypol acetic acid (GA, 12.5 mg∙kg^-1∙d^-1) and desogestrel (DSG, 0.125 mg∙kg^-1∙d^-1)/ethinylestradiol (EE, 0.025 mg∙kg^-1∙d^-1)/ testosterone undecanoate (TU, 100 mg∙kg^-1∙d^-1); group G, a single dose of GA (12.5 mg∙kg^-1∙d^-1) was given; group H, the same dosage of DSG/EE/TU as in group GH were administered; group C, rats were treated with vehicle (1% methyl cellulose) as control. Testes and epididymis were removed at 8 weeks post-treatment for evaluating their weight, volumes, volume fraction, and total volume of testicular tissue structures and the seminiferous tubule diameter using stereological assay. Sperm cell numbers and the motility of epididymal sperm were quantitated by flow cytometry and morphological methods.

Results  Compared with group C, spermatogenesis was normal in group G and suppressed in groups H and GH. Similar changes of testicular tissue structures and sperm number were found in groups H and GH. The decreases of epididymal sperm number and motility in group GH were greater than that of the low-dose gossypol or steroid hormones alone group.

Conclusions  The suppression of spermatogenesis was induced by steroid hormones in the combined regimen, and the epididymis was the target organ of low-dose gossypol. Combined use of low-dose gossypol and steroid hormones played a comprehensive antifertility role in their synergistic effect on reducing the number and motility of epididymal sperm.

     

Drug synergistic antifertility effect of combined administration of low-dose gossypol with steroid hormones in rats

Background Our previous studies suggested that low-dose gossypol combined with steroid hormones has a reversible antifertility role in adult male rats, and the course of treatment was shorter than that of either gossypol or steroid hormones alone. This result suggested that low-dose gossypol and steroid hormones have a drug synergistic effect on antifertility. The aim of the study was to find the target organs of the antifertility synergistic effect of the combined regimen.Methods Thirty-two adult male rats were divided into four groups randomly: group GH, rats were fed orally with group H, the same dosage of DSG/EE/TU as in group GH were administered; group C, rats were treated with vehicle (1% methyl cellulose) as control. Testes and epididymis were removed at 8 weeks post-treatment for evaluating their weight,volumes, volume fraction, and total volume of testicular tissue structures and the seminiferous tubule diameter using stereological assay. Sperm cell numbers and the motility of epididymal sperm were quantitated by flow cytometry and morphological methods.Results Compared with group C, spermatogenesis was normal in group G and suppressed in groups H and GH.Similar changes of testicular tissue structures and sperm number were found in groups H and GH. The decreases of epididymal sperm number and motility in group GH were greater than that of the low-dose gossypol or steroid hormones alone group.Conclusions The suppression of spermatogenesis was induced by steroid hormones in the combined regimen, and the epididymis was the target organ of low-dose gossypol. Combined use of low-dose gossypol and steroid hormones played a comprehensive antifertility role in their synergistic effect on reducing the number and motility of epididymal sperm.     

醋酸棉酚对血浆孕酮和子宫平滑肌的作用

醋酸棉酚能兴奋雌激素化小鼠和豚鼠的离体子宫,对孕激素化或妊娠小鼠和豚鼠的离体子宫无明显影响。给妊娠大鼠灌服醋酸棉酚能使子宫自发收缩明显提高,该作用可被消炎痛拮抗。经醋酸棉酚处理的妊娠大鼠的离体子宫平滑肌对15-Me-PGF_2α的反应比对照组明显提高,醋酸棉酚能降低妊娠大鼠和超排卵大鼠血浆孕酮浓度。结果提示醋酸棉酚兴奋子宫平滑肌作用为雌激素促进;孕激素抑制,促进妊娠大鼠子宫自发收缩可能与其在卵巢水平降低血浆孕酮水平,及促进内源性前列腺素释放有关。     

生精种子汤对男性不育患者精子LDH-x 的影响

本文应用扫描显微分光光度技术对人精子的乳酸脱氢酶同工酶(LDH—X)进行了测定,结果发现:男性不育患者精子的 LDH—X 活性低于有生育力的对照组(P<0.01);经中药生精种子汤治疗后,患者精子的 LDH—X 活性明显升高。提示中药生精种子汤可改善精子的能量代谢,对不育症的治疗有效。