麒麟丸对无精子症模型小鼠睾丸生精功能的影响  

Qilin Pills promote testicular spermatogenesis in azoospermia mice

作　　者：刘纳川 王一洲 姚晨成 赵亮宇[1] 朱子珏[1] 黄煜华[1] 智二磊[1] 万众[1] 陈慧兴[1] 田汝辉[1] 李铮[1] LIU Na-chuan;WANG Yi-zhou;YAO Chen-cheng;ZHAO Liang-yu;ZHU Zi-jue;HUANG Yu-hua;ZHI Er-lei;CHEN Hui-xing;TIAN Ru-hui;LI Zheng(Department of Andrology,Center of Urology / Center for Men＇s Health,Shanghai Jiao Tong University Institute of Urology / Shanghai Key Lab of Reproductive Medicine,Shanghai General Hospital of Shanghai Jiao Tong Universi-ty,Shanghai 200080,China)

机构地区：[1]上海交通大学附属第一人民医院泌尿外科中心男科/上海交通大学泌尿外科研究所男性健康评估中心/上海市生殖医学重点实验室,上海200080

出　　处：《中华男科学杂志》2018年第9期811-818,共8页National Journal of Andrology

基　　金：国家自然科学基金(81671512);上海市申康新兴前沿课题(SHDC12015122)~~

摘　　要：目的:探讨麒麟丸是否促进无精子症模型小鼠生精功能恢复,揭示其调控睾丸生精功能的机制。方法:取15只4周龄ICR雄性小鼠,随机分成模型组、麒麟丸低剂量组、麒麟丸高剂量组,每组5只。采用白消安建立无精子症小鼠模型。建模后麒麟丸组每天给予不同剂量麒麟丸灌胃给药[低剂量组和高剂量组分别为2 000、8 000 mg/(kg·d)],模型组予正常饮食处理,28 d后颈椎脱臼法处死小鼠。HE染色检测小鼠附睾和睾丸组织显微结构,Western印迹检测小鼠睾丸中各级生精细胞、支持细胞和间质细胞特异性标志物表达情况,免疫荧光染色检测睾丸内这些标志物的定位与表达情况。结果:模型组小鼠睾丸中各级生精细胞数量明显减少,大多数附睾管中未见精子; Johnsen评分(5. 2±0. 5)分。麒麟丸高剂量组小鼠生精小管腔内生精细胞排列紧密、层次分明,附睾管腔中见大量精子,未见非精子细胞成分; Johnsen评分(9. 4±0. 6)分。麒麟丸低剂量组小鼠睾丸中各级生精细胞数量相比较模型组有所提升,但是对比高剂量组仍明显减少,部分附睾管中可见精子; Johnsen评分(7. 6±0. 6)分。Johnsen评分模型组显著低于麒麟丸高、低剂量组(P <0. 01),而且麒麟丸高、低剂量组间也有统计学差异(P <0. 05)。Western印迹结果显示,支持细胞标志物GATA4、WT1、SCF、BMP4的表达水平模型组均显著低于麒麟丸高、低剂量组(P <0. 01),高剂量组显著高于低剂量组(P <0. 05或<0. 01);精原干细胞和未分化精原细胞标志物UCHL1、STRA8、NGN3、PLZF 3组小鼠间无显著差异;精母细胞标志物DMC1、SYCP3模型组也均显著低于麒麟丸高、低剂量组(P <0. 05或P <0. 01),高剂量组显著高于低剂量组(P <0. 05或P <0. 01)。免疫荧光染色结果显示,SYCP3的表达与Western印迹结果一致; Ki67荧光信号分布在精原细胞,其荧光信号强度模型组低于麒麟丸高、低剂量组;精子顶体标�Objective： To investigate the effect of Qilin Pills （QP） in facilitating the recovery of spermatogenic function in azoospermia （AS） mice and to explore its mechanism of regulating testicular spermatogenesis. Methods： Fifteen 4-week-old male mice were equally randomized into an AS model control, a low-dose QP and a high-dose QP group. The AS model was established in the mice by intraperitoneal injection of busulfan at 35 mg/kg. After modeling, the animals in the low- and high-dose QP groups were treated with Qilin Pills intragastrically at 2 000 and 8 000 mg/kg/d respectively while those in the model control group fed on a normal diet, all for 28 days. Then, all the mice were sacrificed for examination of the uhrastructures of the epididymis and testis by HE stai-ning, detection of the specific markers of spermatogenic, Sertoli and Leydig cells by Western blot, and determination of the expressions of these markers in the testis tissue by immunofluorescence assay. Results ： The number of spermatogenic cells in the testis tissue was significantly decreased in the AS model eontrols, with no spermatozoa in most of the seminiferous tubules in the epididymis （ Johnsen＇s score ： 5.2 ± 0.5）. In the high-dose QP group, spermatogenic cells were tightly arranged with distinct layers in the seminiferous tu-bules, with a large number of spermatozoa but no non-sperm ceils in the lumens of the epididymis （Johnsen＇s score ： 9.4 ± 0.6）. The number of spermatogenic cells in the testis was increased in the low-dose QP group with some spermatozoa in the seminiferous tubules as compared with that in the model control, but lower than in the high-dose group （Johnsen＇s score： 7.6 ± 0.6）. The Johnsen＇s score was significantly lower in the model control than in the high- and low-dose QP groups （P 〈 0.01 ）, and higher in the high-dose than in the low-dose QP group （P 〈 0.05）. The expressions of the specific markers of Sertoli cells SCF, BMP4, SYCP3, DMC1 and Ki67 were also remarkably lo

关 键 词：麒麟丸 无精子症 白消安 精子发生 支持细胞 小鼠 

分 类 号：R698.2[医药卫生—泌尿科学]