CAJ | 학술논문

目的研究单侧隐睾大鼠对侧睾丸病理变化及SCF/c-kit基因表达,探讨单侧隐睾致对侧睾丸损害的机制.方法 30只SD大鼠随机分为对照组和实验组,实验组复制单侧(左侧)腹腔隐睾模型.3个月后分别取两组右侧睾丸组织进行real-time RT-PCR、Western blot及免疫组化检测干细胞生长因子(SCF)和其受体c-kit基因及其蛋白表达变化,TUNEL法检测细胞凋亡.结果所有动物均存活,与对照组相比实验组对侧睾丸明显缩小,光镜下观察其曲细精管发生退化,生精上皮变薄,管腔较空,生殖细胞明显减少,细胞凋亡增加.两组凋亡指数分别为14.4±0.63和4.45±0.37,差异有统计学意义(P＜0.05).荧光实时定量PT-PCR检测SCF、c-kit基因mRNA含量,实验组对侧睾丸明显降低,两组相比差异有统计学意义(P＜0.05).Western blot检测SCF及c-kit蛋白表达含量,实验组对侧睾丸同样明显降低,两组相比差异有统计学意义(P＜0.05).免疫组化染色显示各级生精细胞膜均有c-kit表达,SCF主要表达于支持细胞膜表面,但实验组两者的表达均较对照组减弱.相关性检验SCF与AI相关系数r=-0.941,P＜0.01;c-kit与AI相关系数r=-0.908,P＜0.01;SCF与c-kit相关系数r=0.956,P＜0.01,均有统计学意义.结论单侧隐睾可致对侧睾丸SCF/c-kit基因表达减弱,生精细胞凋亡增加引起不育.
목적 연구단측은고대서대측고환병리변화급SCF/c-kit기인표체,탐토단측은고치대측고환손해적궤제.방법 30지SD대서수궤분위대조조화실험조,실험조복제단측(좌측)복강은고모형.3개월후분별취량조우측고환조직진행real-time RT-PCR、Western blot급면역조화검측간세포생장인자(SCF)화기수체c-kit기인급기단백표체변화,TUNEL법검측세포조망.결과 소유동물균존활,여대조조상비실험조대측고환명현축소,광경하관찰기곡세정관발생퇴화,생정상피변박,관강교공,생식세포명현감소,세포조망증가.량조조망지수분별위14.4±0.63화4.45±0.37,차이유통계학의의(P＜0.05).형광실시정량PT-PCR검측SCF、c-kit기인mRNA함량,실험조대측고환명현강저,량조상비차이유통계학의의(P＜0.05).Western blot검측SCF급c-kit단백표체함량,실험조대측고환동양명현강저,량조상비차이유통계학의의(P＜0.05).면역조화염색현시각급생정세포막균유c-kit표체,SCF주요표체우지지세포막표면,단실험조량자적표체균교대조조감약.상관성검험SCF여AI상관계수r=-0.941,P＜0.01;c-kit여AI상관계수r=-0.908,P＜0.01;SCF여c-kit상관계수r=0.956,P＜0.01,균유통계학의의.결론 단측은고가치대측고환SCF/c-kit기인표체감약,생정세포조망증가인기불육.
Objective To investigate the pathologic changes and expressions of SCF and c-kit in the contralateral testes in rat model of unilateral cryptorchidism. Methods Thirty male SD rats were maintained under controlled temperature and constant photoperiodic conditions with access to food and water. The rats were randomly assaigned to the control group and the experimental unilateral cryptorchidism group. The left testis of the rats in the unilateral cryptorchidism group was placed into the abdominal cavity. The control group rats were subjected to sham surgery. Three months later, the rats were sacrificed and their right testes were harvested. The pathological changes were observed under microscope. The mRNA and protein expression of SCF and c-kit were also investigated using quantitative Real-time RT-PCR, Western blotting and immunohistochemical staining. Apoptotic germ cells were detected by TUNEL staining. Results All rats survived to the endpoints. The right testes of rats of the unilateral cryptorichid group were smaller than those of the control group. Normal testes of the control group rats manifested active spermatogenesis and orderly arrangement of germ cells.However, disordered and sloughed germinal cells with less distinct seminiferous tubule borders were observed in the contralateral testes of rats with experimental unilateral cryptorichid under the microscope. Cmpared with the control testes, the mRNA and protein expression of SCF and c-kit of the contralateral testes of rats with experimental unilateral cryptorichid was decreased (P＜0.05). The apoptotic germ cells were increased (19.7±3.83 vs 5. 4 ± 1.02, P＜0. 05). The SCF/c-kit expression was positively correlated with the germ cell apoptosis (P＜0. 01 ). Conclusions The decreased expression of SCF and c-kit and increased germ cell apoptosis are found in the contralateral testes, which may contribute to thc infertility in the rat model of experimental unilateral cryptorchidism.