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背景:大量研究证明,一氧化氮合酶/一氧化氮不仅对维持睾丸支持细胞正常功能起重要作用,而且影响精子的发生、活动及受精能力。目的:观察羊睾丸提取液对重金属铅致损小鼠睾丸支持细胞一氧化氮合酶活性的影响。设计:随机对照动物实验。单位:吉林医药学院组织胚胎学教研室。材料:实验于2004-03/2005-08在吉林医药学院组织胚胎学实验室(解放军总后勤部重点实验室)完成。选用健康昆明种雄性小鼠30只,按随机数字表法分为3组,即对照组、睾丸损伤模型组和羊睾丸提取液组,每组10只。方法:睾丸损伤模型组和羊睾丸提取液组小鼠给予100g/L醋酸铅0.2mL/(只·d),5次/周,连续2周,停药1周。羊睾丸提取液组同时腹部皮下注射羊睾丸提取液0.5mL/(只·d)。对照组给予与实验组溶剂等量的重蒸馏水。染毒期满禁食12h后,称体质量后断头处死,解剖取出双侧睾丸,立即称质量,甲醛固定后切片,应用还原型烟酰胺腺嘌呤二核苷酸磷酸-黄递酶组织化学方法结合显微图像分析,观察各组小鼠睾丸支持细胞的一氧化氮合酶活性变化。主要观察指标:染毒期满后各组小鼠的体质量、双侧睾丸质量及睾丸支持细胞一氧化氮合酶吸光度值。结果:30只小鼠全部进入结果分析,无脱失。①染毒期满后羊睾丸提取液组小鼠的体质量、双侧睾丸质量及一氧化氮合酶A值均显著高于睾丸损伤模型组[(22.47±3.49)g,(0.113±0.021)g,0.236±0.020;(19.13±3.46)g,(0.089±0.017)g,0.146±0.023(t=2.151～3.314,P<0.05～0.01)]。②睾丸损伤模型组睾丸一氧化氮合酶阳性支持细胞肿胀变性;羊睾丸提取液组的一氧化氮合酶阳性支持细胞形态接近对照组。结论:羊睾丸提取液可以提高重金属铅致损小鼠睾丸支持细胞的一氧化氮合酶活性,对睾丸损伤有一定的修复和保护作用,可为临床治疗男性不育提供新思路。
BACKGROUND: A large number of studies have demonstrated that nitric oxide synthase/nitric oxide not only plays an important role in maintaining the normal function of testicular Sertoli cells, but also affects the occurrence, activity, and fertilization of sperm. OBJECTIVE: To observe the effect of sheep testicular extract on the nitric oxide synthase activity in mouse testicular Sertoli cells induced by lead-induced heavy metal. Design: Randomized controlled animal experiments. Unit: Department of Histology and Embryology, Jilin Medical College. MATERIALS: The experiment was performed at the Organizational Embryology Laboratory (Key Laboratory of the PLA General Logistics Department) of Jilin Medical College from March 2004 to August 2005. Thirty healthy Kunming male mice were selected and divided into three groups according to the random number table method, namely the control group, the testicular injury model group and the sheep testis extract group, with 10 mice in each group. METHODS: The mice in the testicular injury model group and the sheep testis extract group were given 100g/L lead acetate 0.2mL/(d·d) 5 times per week for 2 weeks and stopped for 1 week. The sheep’s testis extract group was also injected subcutaneously with the sheep’s testicle extract 0.5 mL/(only d). The control group was given an equal amount of double distilled water to the experimental group. After fasting for 12 hours after the expiry of the drug, the body was weighed and was killed by decapitation. The bilateral testis was dissected and immediately weighed. After fixation with formaldehyde, the reduced nicotinamide adenine dinucleotide phosphate diaphorase histochemistry was applied. Methods Combined with microscopic image analysis, the changes of nitric oxide synthase activity in mouse testicular Sertoli cells were observed. MAIN OUTCOME MEASURES: Body weight, bilateral testicular mass, and nitric oxide synthase (NOS) absorbance of testis support cells in the mice after the expiration of the exposure period. RESULTS: All 30 mice were involved in the results analysis without any loss. 1 The body weight, bilateral testis mass, and nitric oxide synthase A were significantly higher in the testes extract group than in the testicular injury model group [(22.47±3.49) g, (0.113±0.021) g, 0.236±0.020; (19.13±3.46) g, (0.089±0.017) g, 0.146±0.023 (t=2.151 to 3.314, P<0.05 to 0.01)]. 2The testicular nitric oxide synthase positive support cells in the testicular injury model group were swollen and degeneration. The positive testicular nitric oxide synthase support cells in the sheep testis extract group were similar to the control group. Conclusion: The sheep’s testis extract can improve the nitric oxide synthase activity of testis supporting cells in mice with lead-induced damage by heavy metals. It has certain repair and protection effects on testicular injuries and can provide new ideas for clinical treatment of male infertility.