顺铂染毒大鼠睾丸定量组织学分析及睾丸酶活力的变化  

Quantitative histological analysis of testes and effect on testicular enzymes induced by cisplatin in rats

作　　者：孙铱钒 关朕[1] 孙应彪[1] 苏莉[1] 朱安[1] 常旭红[1] 刘志飞[1] 郭芳[1] 李晋[1] 

机构地区：[1]兰州大学公共卫生学院,甘肃兰州730000

出　　处：《毒理学杂志》2015年第3期172-176,共5页Journal of Toxicology

基　　金：国家自然科学基金(81402703);国家大学生创新训练项目(821003153);中央高校基本科研业务费专项资金(lzu-jbky-2015-182)

摘　　要：目的探讨顺铂(CP)染毒大鼠睾丸组织病理学和睾丸酶活力的变化。方法选择健康成年雄性Wistar大鼠,随机分为CP 1.0、2.5和5.0 mg/kg组及对照组,连续腹腔注射染毒3 d。染毒结束后,制备PAS(Periodic AcidSchiff)染色病理切片,观察睾丸组织病理变化并行定量组织学分析。分光光度法测定琥珀酸脱氢酶(SDH)、苹果酸脱氢酶(MDH)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、诱导型一氧化氮合酶(i NOS)和Na+-K+-ATPase活力。实时荧光定量PCR检测i NOS和Na+-K+-ATPasemRNA表达水平。结果 CP染毒后大鼠睾丸组织病理学改变主要见于睾丸生精上皮精子发生周期第Ⅱ-Ⅲ期。定量组织学分析发现,在生精上皮第Ⅱ-Ⅲ期CP 2.5和5.0 mg/kg组大鼠睾丸生精小管相对面积和直径变小,管腔相对面积增加(P<0.05)。CP各剂量组生精上皮占生精小管面积百分比和生精小管管腔直径均变小(P<0.05),CP5.0 mg/kg组睾丸间质血管面积增加(P<0.01)。与对照组比较,CP 5.0 mg/kg组精子数减少,精子活力降低(P<0.05)。CP各染毒组SDH活力均低于对照组,而MDH和i NOS活力仅5.0 mg/kg组低于对照组(P<0.05)。CP 5.0 mg/kg组ACP、AKP和Na+-K+-ATPase活力均明显高于对照组(P<0.05)。RT-PCR结果显示,Na+-K+-ATPase和i NOS mRNA表达水平与酶活力改变一致。结论顺铂可能参与改变睾丸细胞能量代谢相关酶活力而致睾丸细胞损伤。Objective To investigate the effects of cisplatin（ CP） on the changes of testicular tissue pathology and enzyme activities in rats. Methods Mature male Wistar rats were randomly divided into four groups： control group,and CP 1. 0 mg / kg,2. 5 mg / kg,5. 0mg / kg groups. Rats were intoxicated by intraperitoneal injection once a day for three consecutive days,respectively. Periodic Acid-Schiff staining was used to observe the changes of testicular tissue pathology with quantitative histological analysis. The activities of succinate dehydrogenase（ SHD）,malate dehydrogenase（ MDH）,acid phosphatase（ ACP）,alkaline phosphatase（ AKP）,inducible nitric oxide synthase（ i NOS） and Na＋-K＋-ATPase were detected by spectrophotometry. The levels of i NOS and Na＋-K＋-ATPase mRNA were measured with real-time qantitive PCR. Results Pathological examination showed that the changes of testicular tissue pathology mainly emerged in Ⅱ-Ⅲ stage of seminiferous epithelium cycle in rats. Quantitative histological analysis showed that relative area and diameter of seminiferous tubule were smaller than control group,but the relative lumen area of seminiferous tubule increased in CP 2. 5 mg / kg and 5. 0 mg / kg groups（ P〈0. 05）. The ratio of seminiferous epithelium and seminiferous tubule area as well as lumen diameter of seminiferous tubule reduced in all exposure groups（ P〈0. 05）,but the vascular area of testicular interstitium increased in CP 5. 0 mg / kg group in Ⅱ-Ⅲ stage of seminiferous epithelium cycle（ P〈0. 01）. The spermatozoa density and sperm motility were reduced in CP 5. 0 mg / kg group（ P〈0. 05）.CP inhibited SDH activity in all exposure groups as well as MDH and i NOS in CP 5. 0 mg / kg group（ P〈0. 05）. The activities of ACP,AKP and Na＋-K＋-ATPase obviously increased in CP 5. 0mg / kg group（ P〈0. 05）. The levels of i NOS and Na＋-K＋-ATPase mRNA expression were consistent with its enzyme activity. Conclusion CP-inhibited the activity of testicular en

关 键 词：顺铂 定量组织学分析 睾丸酶 

分 类 号：R996.3[医药卫生—毒理学]