GYY4137对精索静脉曲张大鼠附睾上皮细胞凋亡的影响  

Effect of GYY4137 on epididymal epithelial cell apoptosis in rats with varicocele

作　　者：夏煜琦 程帆[1] 宁金卓[1] 余伟民[1] 饶婷[1] 阮远[1] XIA Yuqi;CHENG Fan;NING Jinzhuo;YU Weimin;RAO Ting;RUAN Yuan(Department of Urology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院泌尿外科

出　　处：《中华实用诊断与治疗杂志》2019年第11期1044-1048,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：中央高校基本科研业务专项资金(青年教师资助项目)(413000227)

摘　　要：目的探讨缓释硫化氢供体GYY4137对精索静脉曲张(varicocele,VC)大鼠附睾上皮细胞凋亡的影响及可能机制。方法雄性SD大鼠60只,随机分为假手术组、假手术+GYY4137组、VC组、VC+GYY4137组各15只。VC组与VC+GYY4137组结扎左肾静脉制备VC模型,假手术组、假手术+GYY4137组仅开腹不结扎左肾静脉。VC+GYY4137组、假手术+GYY4137组术后腹腔注射GYY4137(20mg/kg GYY4137溶于1mL生理盐水),1次/d,连续4周,VC组、假手术组腹腔注射等量生理盐水。药物处理4周后处死大鼠,取左侧附睾组织行病理检查,TUNEL法检测4组附睾上皮细胞凋亡指数(apoptotic index,AI),免疫组织化学法检测附睾上皮细胞细胞色素C(cytochrome C,CytC)、caspase-3阳性表达率,Western blot法检测附睾组织CytC、caspase-3蛋白相对表达量,实时荧光定量PCR法检测附睾组织CytC mRNA、caspase-3mRNA相对表达量。结果药物处理4周,假手术组、假手术+GYY4137组附睾结构正常、上皮细胞排列整齐,VC组附睾组织结构萎缩、附睾管腔结构破坏、间质水肿,VC+GYY4137组附睾结构破坏、间质水肿程度较VC组轻;药物处理4周,VC组、VC+GYY4137组附睾上皮细胞AI[(23.18±2.09)%、(12.73±1.14)%]、CytC阳性表达率[(25.24±1.99)%、(13.88±1.26)%]及caspase-3阳性表达率[(28.20±2.11)%、(17.33±1.28)%]高于假手术组[(5.46±0.46)%、(5.37±0.77)%、(4.52±0.63)%]、假手术+GYY4137组[(5.86±0.55)%、(5.53±0.52)%、(4.93±0.65)%](P<0.05),VC组高于VC+GYY4137组(P<0.05),假手术组与假手术+GYY4137组比较差异无统计学意义(P>0.05);VC组、VC+GYY4137组附睾组织CytC蛋白相对表达量(0.38±0.02、0.27±0.01)、CytC mRNA相对表达量(3.64±0.41、1.55±0.14)、caspase-3蛋白相对表达量(0.27±0.02、0.22±0.01)、caspase-3mRNA相对表达量(4.09±0.36、2.27±0.32)均高于假手术组(CytC蛋白:0.11±0.03;CytC mRNA:1.01±0.04;caspase-3蛋白:0.10±0.01;caspase-3mRNA:1.03±0.15)、假手术+GYY4137组(CytC蛋白:0.12±0.01;CytC mObjective To investigate the effect of GYY4137,a new hydrogen sulfide donor,on the epididymal epithelial cell apoptosis in rats with varicocele(VC)and its possible mechanism.Methods Sixty male rats were randomly divided into 4 groups:sham operation group,sham operation+GYY4137 group,VC group and VC+GYY4137 group,with 15 rats in each group.The left renal vein was ligated in VC group and VC+GYY4137 group to prepare the VC model,while the abdominal cavity was opened without ligating the left renal vein in sham operation group and sham operation+GYY4137 group.VC+GYY4137 group and sham operation+GYY4137 group received intraperitoneal injection of GYY413720 mg/kg dissolved in 1 mL normal saline,once a day,totally for 4 weeks.While in VC group and sham operation group,1 mL normal saline was intraperitoneally injected.After 4 weeks,the rats were sacrificed and epididymal tissue was taken for HE staining and TUNEL assay to observe pathological damage and measure the apoptotic index of epididymal epithelial cells.The positive rates and relative expression levels of CytC and caspase-3 proteins were detected by immunohistochemistry and Western blot.The relative expression levels of CytC and caspase-3 mRNA were detected by real-time quantitative PCR.Results After 4 weeks of modeling,the epididymal tissues in sham operation group and sham operation+GYY4137 group showed normal epididymal structure and orderly epithelial cells,while atrophy of epididymal structure,destruction of epididymal lumen structure and edema in VC group.In VC+GYY4137 group,the damages of epididymal structure and edema were mild.The apoptotic indexes of epididymal epithelial cells((23.18±2.09)%,(12.73±1.14)%),CytC positive rates((25.24±1.99)%,(13.88±1.26)%)and caspase-3 positive rates((28.20±2.11)%,(17.33±1.28)%)in VC group and VC+GYY4137 group were significantly higher than those in sham operation group((5.46±0.46)%,(5.37±0.77)%,(4.52±0.63)%)and sham operation+GYY4137 group((5.86±0.55)%,(5.53±0.52)%,(4.93±0.65)%)(P<0.05),and higher in VC group

关 键 词：精索静脉曲张 GYY4137 附睾上皮细胞凋亡 细胞色素C CASPASE-3 大鼠 

分 类 号：R73[医药卫生—肿瘤]