睾丸动脉血流障碍对小鼠睾丸生精上皮和血－睾屏障的影响

Effect of unilateral testicular artery ligating/reperfusion on seminiferous epithelium and blood-testis barrier in mice

目的：探讨睾丸动脉血流障碍对小鼠睾丸生精上皮和血-睾屏障的影响.方法：成熟雄性BALB/c小鼠,随机分为对照组和6个实验组,对照组仅暴露睾丸动脉而不进行结扎,实验组显微镜下行单侧睾丸动脉结扎, H-E染色观察睾丸组织的病理改变,电镜观察血-睾屏障的变化.结果：对照组睾丸生精小管无明显病理改变;1h组睾丸生精小管上皮细胞排列不均匀,细胞界限清楚,线粒体轻微肿胀,核周隙轻微增宽,睾丸生精小管的基膜完整;2h和3h组生精小管生精细胞层数减少,线粒体轻微空泡改变,线粒体嵴扩张,支持细胞内质网扩张明显,有局部液化灶,生精小管基膜轻微波纹状;睾丸间质可见炎性细胞浸润,多为分叶的中性粒细胞;4h和5h组睾丸间质血管中充血明显,血管内壁有大量的炎性细胞贴壁现象,血管壁增厚呈玻璃样变,精原细胞有核碎裂现象,细胞界限不清,基膜间隙增宽,基膜皱折,精子细胞顶体位置未见顶体,精子细胞局部有空泡,界限不清,细胞膜不完整;6h组睾丸生精小管内主要为精原细胞和初级精母细胞,精原细胞与基膜脱离,基膜有＂分层＂现象,支持细胞、精母细胞肿胀明显,细胞膜不完整,界限不清,生精小管内可见多核巨细胞.结论：单侧睾丸动脉结扎/再通可引起睾丸生精细胞的缺血性损伤和坏死,生精上皮细胞和血-睾屏障的病理变化随缺血时间的延长而加重.

Objective： To explore the effect of testicular artery hemodynamic disorder on seminiferous epithelium and blood- testis barrier in mice. Methods： Sixty male BALB/e mice were euthanized under chlorine hydrate and were randomly divided into seven groups.. ①control group：the mice were treated with sham operation （no ligating testicular artery） ;②experimental group： six subgroups were established （reperfusion for 1 h, 2 h, 3 h, 4 h, 5 h, 6 h after undergoing ligation of testicular artery）. The mice were killed 12 h after operation. Pathological change in testicular tissue was observed using hematoxylin and eosin （HE） staining and blood-testis barrier change was investigated by electron microscope. Results： No obvious patho- logical change with seminiferous tubule had been detected in the control group, epithelial cell arrangement was disordered, cytochondriome was swelled, perinuclear cisterna was widened and basement membrane was maintained integrity in 1 h experimental group. Pathologically, less layers and disorder inarrangement of spermatogenic cells, extension of mitochondrial crista, expansion of sustentacular cell endoplasmic reticulum, microwave stripe-like change of seminiferous tubule basement membrane and infiltrating inflammatory cells （sublobe neutrophilic granulocyte） in interstitial tissues were found in 2 h and 3 h expermental groups. The blood vessel hyperemiaed in the interstitial tissues of the testis, a great quantity inflammatory cells were adhered to the blood vessel inner wall, blood vessel inner wall was thickened, spermatogonium developed nuclear fragmentation, and spermatid apicle and bulla disappeared in 4 h and 5 h experimental groups. In 6 h experimental group, abundant spermatogonium and primary spermatocyte were observed in the seminiferous tubule. Germ cells were separated from the basal membrane, spermatocytes were obviously swelled, the cell membrane was absent, and Warthin-Finkeldey cells generated in the seminiferous tubule. Conclusion： Unilateral ligation/reperfusion of the testicular artery cause spermatogenie cell ischemic injury and necrosis. The pathological change of seminiferous epithelium cells and blood-testis barrier is aggratating gradually with prolongation of ischemic times.