急性热应激对性成熟雄性小鼠睾丸、附睾、输精管中热休克蛋白70表达的影响

目的:探讨急性热应激对性成熟雄性小鼠睾丸、附睾、输精管中热休克蛋白70(heat shock protein 70,HSP70)表达的影响。方法:将32只8周龄雄性小白鼠随机均分为4组,饲养7d后,进行热应激处理,温度控制在(39±0.5)℃,时间分别为0.5、1和3h。应激后立即采血,分离血清测定谷草转氨酶(GOT)含量。一侧附睾制备精子悬液,用于计算精子密度和顶体畸形率;另一侧附睾、睾丸、输精管用于免疫组化研究。结果:应激后,小鼠体重、睾丸系数、顶体畸形率变化不显著(P>0.05),附睾系数和精子密度有不同程度的下降,GOT含量急剧升高(P<0.01)。随着应激时间的延长,小鼠精子密度呈递减趋势,顶体畸形率呈上升趋势。应激时间最短的0.5h组小鼠体重、睾丸系数、附睾系数的降幅反而最大。免疫组化法观察发现,HSP70在性成熟小鼠睾丸、附睾、输精管中均有表达。正常状态下,HSP70在睾丸组织间质细胞中少量表达,应激后分布于间质细胞核,此外在精母细胞核与精子细胞核中也有大量分布;附睾中HSP70主要分布于主细胞质,基细胞和亮细胞中没有表达,应激后附睾体的纤毛细胞中也发现大量棕色颗粒;输精管中HSP70主要定位在基细胞质,主细胞中不表达。随着应激时间的延长,HSP70在睾丸、附睾中的表达量明显升高,而在输精管中的增幅不明显。结论:急性热应激对性成熟雄性小鼠的生殖系统造成了损伤;HSP70在睾丸、附睾、输精管中的表达与定位具有区域特异性和细胞特异性,提示其可能参与精子的发生与成熟;HSP70在应激状态下表达量大幅上升的作用可能在于保护细胞免受高热损伤。     

Multiple effects of sibutramine on ejaculation and on vas deferens and seminal vesicle contractility

Sibutramine is an inhibitor of norepinephrine and 5-HT reuptake largely used in the management of obesity. Although a fairly safe drug, postmarketing adverse effects of sibutramine were reported including abnormal ejaculation in men. This study investigates the effects of sibutramine on ejaculation and vas deferens and seminal vesicle contractility. Adult male rats received sibutramine (5; 20; or 50 mg kg^− 1, ip) and after 60 min were exposed to receptive females for determination of ejaculation parameters. The vasa deferentia and seminal vesicles of untreated rats were mounted in isolated organ baths for recording of isometric contractions and HEK293 cells loaded with fluorescent calcium indicator were used to measure intracellular Ca²⁺ transients. Sibutramine 5 and 20 mg kg^− 1 reduced ejaculation latency whereas 50 mg kg^− 1 increased ejaculation latency. Sibutramine 3 to 30 μM greatly increased the sensitivity of the seminal vesicle and vas deferens to norepinephrine, but at concentrations higher than 10 μM there were striking depressions of maximal contractions induced by norepinephrine, carbachol and CaCl₂. In HEK293 cells, sibutramine 10 to 100 μM inhibited intracellular Ca²⁺ transients induced by carbachol. Depending on the doses, sibutramine either facilitates or inhibits ejaculation. Apart from its actions in the central nervous system, facilitation of ejaculation may result from augmented sensitivity of smooth muscles to norepinephrine while reductions of intracellular Ca²⁺ may be involved in the delayed ejaculation observed with high doses of sibutramine.     

Reinnervation of transplanted vas deferens by cholinergic nerves normally supplying skeletal muscle

The rat vas deferens was removed and either transplanted alongside the soleus muscle or into the bed of the soleus muscle that had previously been removed, and in this case the soleus nerve was connected to the transplant. The vas deferens reinnervated by the somatomotor nerve recovered the best. Contractions to transmural electrical stimulation could not be elicited from the denervated vas deferens, although noradrenaline and acetylcholine elicited contractions. The reinnervated vas deferens produced good contractile responses to transmural stimulation, and these were substantially reduced by a cholinergic muscarinic blocking agent, hyoscine, as compared to only a small reduction in the control vas deferens. Neostigmine potentiated the contraction of the transplanted vas deferens to a greater extent than that of the control. This indicated that a substantial component of the contractile response was produced by cholinergic fibres. Consistent with this was the finding that, while guanethidine blocked a greater proportion of the contraction in the control vas deferens, the contraction of the reinnervated transplant was less affected. Acetylcholine elicited a strong contraction in control vas deferens, but only a small response was obtained in the reinnervated transplant. However, the response to noradrenaline was greater in the transplant than in the control vas deferens. These results indicate that cholinergic nerves normally supplying skeletal muscle can reinnervate smooth muscle and that the alien somatomotor innervation altered the responsiveness of the smooth muscle of the vas deferens. Morphological studies confirm the shift from adrenergic to cholinergic fibres in the reinnervated vas deferens.     

The peritoneal cavity as a bioreactor for tissue engineering visceral organs: bladder, uterus and vas deferens

Our objective was to produce avascular, myofibroblast-rich tissue capsules for use as autologous grafts for hollow, smooth muscle-walled visceral organs-bladder, uterus and vas deferens. To produce tissue for grafting, templates of the appropriate shape were implanted in the peritoneal cavities of rats or rabbits. After 2-3 weeks, the templates were removed, the encapsulating myofibroblast-rich tissue harvested and grafted to replace resected segments of bladder, vas deferens or uterus of the same animals in which the tissue was grown. Bladder grafts showed 100% patency after 14 months and had developed a morphology similar to normal bladder. Tubes of myofibroblast tissue grafted unilaterally into resected rabbit vasa deferentia developed a morphology resembling native tissue, with sperm in the ejaculate indicative of normal function. At 12 weeks after grafting, uterine graft tissue had increased in thickness and developed the morphology of normal uterus, with endometrium overlying several layers of smooth muscle cells (myometrium-like) which were interspersed with collagen fibrils; grafted uterine horns supported embryos to the late stages of gestation. This study shows that myofibroblast tissue produced in the peritoneal cavity is sufficiently plastic to permit differentiation of cells into bladder, vas deferens or uterine smooth muscle. As a method for producing autologous graft material for repair/replacement of these organs, this approach has many benefits over conventional and current tissue-engineering strategies.     

The effects of lignocaine on actions of the venom from the yellow scorpion “Leiurus quinquestriatus” in vivo and in vitro

Fatani, A.J., Harvey, A.L., Furman, B.L., and Rowan, E.G. The effects of lignocaine on actions of the venom from the yellow scorpion, Leiurus quinquestriatus, in vivo and in vitro. Toxicon, 19. Many toxins from scorpion venoms activate sodium channels, thereby enhancing neurotransmitter release. The aim of the present work was to determine if the in vivo and in vitro effects of Leiurus quinquestriatus venom (LQQ) could be ameliorated by lignocaine, a sodium channel blocker. In urethane anaesthetised rabbits, LQQ venom (0.5 mg kg⁻¹, i.v.) caused initial hypotension and bradycardia followed by hypertension, pulmonary oedema, electrocardiographic changes indicating conduction defects, ischaemia, infarction, and then hypotension and death. Lignocaine (1 mg kg⁻¹ i.v. bolus initially, followed by i.v. infusion of 50 μg kg⁻¹ min⁻¹) significantly attenuated the majority of the venom-evoked effects and reduced mortality. Addition of LQQ venom (1, 3 and 10 μg ml⁻¹) to chick biventer cervicis, guinea pig ileum, and rat vas deferens preparations, increased the height of electrically-induced twitches, elevated resting tension, and caused autorhythmic oscillations. Lignocaine (3 × 10⁻⁴–1.2 × 10⁻³ M) greatly attenuated these venom-evoked actions in the three preparations. Antagonists of appropriate neurotransmitters were also tested to determine the contribution of released transmitters to LQQ effects. Atropine significantly decreased the venom-elicited effects on guinea pig ileum preparations, while prazosin and guanethidine significantly reduced the venom’s actions on rat vas deferens. In chick biventer cervicis preparations, tubocurarine and hexamethonium significantly attenuated the venom-induced effects. This study supports the hypothesis that many effects of LQQ venom involve the release of neurotransmitters and may be ameliorated by treatment with lignocaine.     

紫贻贝水提取物对大鼠离体子宫、平滑肌的作用

本文首次报道了紫贻贝水提取液对大鼠离体子宫、平滑肌的作用及其作用机制。结果表明:①贻贝水提取液对大鼠离体子宫有明显收缩作用,对场刺激输精管引起的收缩有明显增强作用,而这些收缩作用可被α—受体阻断剂酚妥拉明所对抗与阻断。②对大鼠离体胃条与回肠平滑肌有松弛作用,而这种松弛作用也可被α—受体阻断剂酚妥拉明所对抗与阻断。③对蟾蜍血管灌注则流量减少从47±11dr/min降至24±7dr/min,(P<0.05),故表明紫贻贝水提取液对大鼠离体子宫呈收缩作用,而这种作用是由于兴奋了α—受体的结果。     

输精管逆向注射30%乙醇引起大鼠精子活力低下的实验研究

目的:探讨输精管逆向注射30%乙醇至附睾导致大鼠精子活力低下的原因以及机制。方法:40只雄性SD大鼠随机分为注射组(n=15)、假手术对照组(n=15)和正常对照组(n=10),注射组在输精管靠近附睾2 cm处用1 m l注射器注射30%乙醇0.5 m l;假手术组只在输精管管壁相同部位用注射针刺一小孔;对照组未作任何处置。术后1个月,比较3组大鼠精子活力,苏木精-伊红(HE)染色观察输精管管壁的变化,检测附睾组织匀浆中白细胞介素6(IL-6)、γ干扰素(IFN-γ)和肉碱的含量变化。结果:1个月后,注射组的精子活动率为(30±14)%,假手术组(64±11)%,正常组为(68±9)%。注射组精子活力明显降低,与假手术对照组和正常对照组比较差异有极显著性(P<0.01)。IL-6、IFN-γ含量注射组分别为(772.7±211.0)、(350.7±39.0)pg/m l,假手术组分别为(308.5±121.0)、(172.2±61.0)pg/m l,正常对照组分别为(287.8±143.0)、(163.8±21.0)pg/m l,注射组显著高于假手术组和对照组(P<0.05)。注射组肉碱含量[(491.1±48.0)mol/L]显著低于假手术组[(664.6±45.0)mol/L]和对照组[(605.5±99.0)mol/L],P均<0.05。结论:输精管逆向注射低浓度乙醇能干扰附睾中的精子成熟环境,并激活生殖系统的免疫细胞分泌细胞因子,导致精子活力低下。     

Intracytoplasmic sperm injection using surgically retrieved epididymal and testicular spermatozoa in cases of obstructive and non-obstructive azoospermia

This was a retrospective study of 115 patients who underwent 124 cycles of ICSI using surgically retrieved spermatozoa. The objective was to compare the results of ICSI in patients with obstructive azoospermia using epididymal spermatozoa (36 cycles) or testicular spermatozoa (58 cycles) with ICSI in patients with non-obstructive azoospermia using testicular spermatozoa (30 cycles). When epididymal spermatozoa were used for ICSI, the fertilization rate per injected metaphase-II oocyte and the clinical pregnancy rate per ICSI cycle were 60.4 and 25%, respectively. When testicular spermatozoa were used in obstructive cases, the fertilization rate and pregnancy rate were 57.9 and 34.5%. In non-obstructive cases the fertilization and pregnancy rates were 41.2 and 16.6%. When patients with obstructive azoospermia were regrouped according to the cause of obstruction, the fertilization and pregnancy rates were 59.1 and 35.1% in acquired obstruction and 58.7 and 24.3% in congenital obstruction. The fertilization and pregnancy rates were not statistically different ( p  > 0.05) when testicular or epididymal spermatozoa were used in obstructive cases; neither was statistically different ( p  > 0.05) when compared in patients with congenital and acquired obstruction. On the other hand, the fertilization and pregnancy rates in cases with non-obstructive azoospermia were significantly lower ( p  < 0.05) than in obstructive cases.     

苯乙烯－马来酸酐聚合物的抗生育研究

采用自行研制的两批苯乙烯－马来酸酐（ＳＭＡ）聚合物，即ＳＭＡ－Ａ和ＳＭＡ－Ｃ进行抗生育实验。结果表明，大白鼠每侧输精管注入ＳＭＡ２．５ｍｇ，一年内８５～９０％的实验动物通而不育。雄兔每侧输精管注入ＳＭＡ３７．５ｍｇ，可造成阻塞性无精子。实验动物无不良反应。     

1-(α-萘甲基)2-甲基-6,7-二甲氧基-四氢异喹啉对大鼠α-肾上腺素受体的阻断作用

1—(α—萘甲基)2—甲基—6,7—二甲氧基—四氢异喹啉;;α—肾上腺素受体;;肛尾肌输精管