离体睾丸缺血再灌注损伤的实验研究

目的 探讨人离体睾丸缺血再灌注损伤的变化规律。了解益生注射液对其变化的干预效果。方法 采用13人摆脱赠尸睾丸26只，用4℃250ml高渗枸橼酸盐嘌呤液冷保存，再以此液500ml再灌注，实验组用含500μg/ml益生注射液同法操作，同一睾丸于单纯冷保存（对照组睾丸8只，实验组8只）6、12、18、24、36、48、60和72小时连续取材，冷保存6、12、18、24和36小时（对照组与实验组每时间点各1只，共10只）后，37）复温再灌注6和12小时连续取材，作组织学及酶组织化学染色。结果 实验组睾丸组织4℃冷保存24小时内无明显病理改变；36和48小时后出现精索血管内皮细胞肿胀，变性及部分脱落，曲细精管与生精上皮剥离，间质轻度水肿等损伤；冷保存12小时乳酸脱氢酶与琥珀酸脱氢酶活性增高，24小时后又降低；一氧化氮合成酶活性于冷保存18小时后降低再灌注后组织损伤加重，对照组睾丸单纯冷保存12小时即出现精索血管内皮细胞肿胀，变性，24小时后加重，再灌注后精索血管大量内皮细胞变性脱落，曲细精管基膜剥离和间质水肿严重。结论 4℃冷保存24小时内仅有内皮细胞轻微损伤，超过24小时可引起睾丸血管及组织形态结构明显异常，37℃复温再灌注使损伤程度加重，益生注射液使人离体睾丸在4℃冷保存24小时内保持正常形态结构。且对再灌注损伤有保护作用。

EXPERIMENTAL STUDY ON ISOLATED TESTES WITH ISCHEMIA/ REPERFUSION INJURY

OBJECTIVE: To investigate the injury on isolated testes induced by ischemia/reperfusion(I/R), and the protective effect of Yisheng injection on the injury. METHODS: Twenty-six isolated cadaver testes contributed by 13 persons were preserved with 4 degrees C 250 ml hypertonic citrate alloxuric (HCA) solution and then reperfused with 37 degrees C 500 ml HCA. Solution of experimental group contained 500 micrograms/ml Yisheng injection. In simple cold preservation test, involving in 8 experimental and 8 control testes, a series of time points (6, 12, 18, 24, 36, 48, 60, 72 hours) were set to harvest. 10 testes (1 testis respectively on 6, 12, 18, 24 and 36 hours in experimental and control groups) were reperfused with 37 degrees C HCA for 6 and 12 hours. Histological and histochemical changes were observed. RESULTS: In the experimental testes, 4 degrees C cold preservation in 24 hours could not induce obvious pathologic changes. After 24 hours, changes such as swelling, vacuolar degeneration or detachment of endothelial cells (ECs), separation between basement membrane and seminiferous epithelium, mal-alignment of spermatogenous cell and edema of mesenchyme could be observed. In the testes preserved for 12 hours, the activity of lactic dehydrogenase(LDH) and succinic dehydrogenase (SDH) increased, then fallen after 24 hours. The activity of Nitric oxide synthetase(NOS) decreased after 18 hours. All changes were more obvious after following 37 degrees C reperfusion. In the control testes, swelling and vacuolar degeneration of ECs occurred on 12 hours cold preservation, and injury was worse along with the prolongation of cold preservation time. Pathologic changes of ECs, seminiferous epithelium and mesenchyme were serious after 37 degrees C reperfusion. CONCLUSION: 4 degrees C cold preservation in 24 hours can only cause mild ECs' injury, and obvious abnormal testes' histological profile can be observed beyond 24 hours. 37 degrees C reperfusion will make injury worse. Yisheng injection can keep isolated testes histologic structure well in 24 hours cold preservation, and it has protective effect on I/R injury.