睾丸去神经支配经由caspase-8通路诱导Leydig细胞凋亡

发布时间：2018-05-20 21:04

本文选题：睾丸神经 + Leydig细胞　；参考：《南京医科大学》2009年硕士论文

【摘要】：目的研究睾丸去神经支配对Leydig细胞凋亡的影响及其分子机制。方法健康成年雄性SD大鼠80只,随机分为精索上神经切除假手术组(SSG)、精索上神经切除组(SDG)、精索下神经切除假手术组(ISG)和精索下神经切除组(IDG),解剖显微镜下建立睾丸去神经支配大鼠模型,于术后14天、21天各组均取10只大鼠处死,取双侧睾丸及血液标本,采用脱氧核糖核酸末端转移酶介导的缺口末端标记技术(TUNEL)、透射电镜及Annexin V-FITC/PI双标流式细胞术检测Leydig细胞的凋亡,放射性免疫技术检测血清中睾酮(T)、卵泡刺激素(FSH)和黄体生成素(LH)水平,免疫荧光技术及荧光激活细胞分选术(FACS)检测Leydig细胞caspase-3, 8, 9表达。结果精索上神经或精索下神经切除后14天和21天,TUNEL与Annexin V-FITC/PI双标流式细胞术显示大鼠睾丸中凋亡的Leydig细胞显著增多,电镜分析显示Leydig细胞出现染色质凝聚、细胞皱缩和凋亡小体形成等凋亡特征性超微结构改变;术后14天、21天T水平进行性下降,LH水平手术后持续升高, FSH水平无显著改变;免疫荧光术和FACS进一步显示切除精索神经后,caspase-3和caspase-8的表达与活性在Leydig细胞中显著增加,而caspase-9表达与活性无显著改变。结论睾丸支配神经是Leydig细胞重要的生存因子,切除该类神经将经由caspase-8介导的死亡受体通路诱发Leydig细胞凋亡。
[Abstract]:Purpose To study the effect of testicular denervation on apoptosis of Leydig cells and its molecular mechanism. Method 80 healthy adult male SD rats, The rat models of testicular denervation were established under anatomic microscope. The rats were randomly divided into three groups: the supraspermatic nerve resection group (SSG), the superior spermatic nerve resection group (SDG), the subseminal nerve resection group (ISG) and the subseminal nerve resection group (IDGN), and the testicular denervated rat model was established under anatomic microscope. Ten rats in each group were killed on the 14th day and 21st day after operation. Bilateral testis and blood samples were collected. The apoptosis of Leydig cells was detected by means of terminal deoxyribonucleic acid transferase-mediated Nick end labeling (Tunel), transmission electron microscopy (TEM) and Annexin V-FITC/PI double labeled flow cytometry. The levels of testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) in serum were detected by radioimmunoassay, and the expression of caspase-3,8,9 in Leydig cells was detected by immunofluorescence and fluorescence activated cell sorting. Result Tunel and Annexin V-FITC/PI double labeled flow cytometry showed that the number of apoptotic Leydig cells in testis of rats was significantly increased at 14 and 21 days after superior or inferior spermatic nerve resection. Electron microscopic analysis showed that Leydig cells showed chromatin condensation. The ultrastructural changes of apoptosis such as cell shrinkage and formation of apoptotic corpuscles continued to increase after 14 days and 21 days after operation, but the level of FSH did not change significantly. Immunofluorescence and FACS showed that the expression and activity of caspase-3 and caspase-8 increased significantly in Leydig cells after spermatectomy, but caspase-9 expression and activity did not change significantly. Conclusion Testicular innervation is an important survival factor for Leydig cells. Excision of these nerves will induce apoptosis of Leydig cells via caspase-8 mediated death receptor pathway.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R346

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