本文選題：睪丸神經(jīng) + Leydig細(xì)胞�。� 參考：《南京醫(yī)科大學(xué)》2009年碩士論文

【摘要】：目的 研究睪丸去神經(jīng)支配對(duì)Leydig細(xì)胞凋亡的影響及其分子機(jī)制。 方法 健康成年雄性SD大鼠80只,隨機(jī)分為精索上神經(jīng)切除假手術(shù)組(SSG)、精索上神經(jīng)切除組(SDG)、精索下神經(jīng)切除假手術(shù)組(ISG)和精索下神經(jīng)切除組(IDG),解剖顯微鏡下建立睪丸去神經(jīng)支配大鼠模型,于術(shù)后14天、21天各組均取10只大鼠處死,取雙側(cè)睪丸及血液標(biāo)本,采用脫氧核糖核酸末端轉(zhuǎn)移酶介導(dǎo)的缺口末端標(biāo)記技術(shù)(TUNEL)、透射電鏡及Annexin V-FITC/PI雙標(biāo)流式細(xì)胞術(shù)檢測(cè)Leydig細(xì)胞的凋亡,放射性免疫技術(shù)檢測(cè)血清中睪酮(T)、卵泡刺激素(FSH)和黃體生成素(LH)水平,免疫熒光技術(shù)及熒光激活細(xì)胞分選術(shù)(FACS)檢測(cè)Leydig細(xì)胞caspase-3, 8, 9表達(dá)。 結(jié)果 精索上神經(jīng)或精索下神經(jīng)切除后14天和21天,TUNEL與Annexin V-FITC/PI雙標(biāo)流式細(xì)胞術(shù)顯示大鼠睪丸中凋亡的Leydig細(xì)胞顯著增多,電鏡分析顯示Leydig細(xì)胞出現(xiàn)染色質(zhì)凝聚、細(xì)胞皺縮和凋亡小體形成等凋亡特征性超微結(jié)構(gòu)改變;術(shù)后14天、21天T水平進(jìn)行性下降,LH水平手術(shù)后持續(xù)升高, FSH水平無顯著改變;免疫熒光術(shù)和FACS進(jìn)一步顯示切除精索神經(jīng)后,caspase-3和caspase-8的表達(dá)與活性在Leydig細(xì)胞中顯著增加,而caspase-9表達(dá)與活性無顯著改變。 結(jié)論 睪丸支配神經(jīng)是Leydig細(xì)胞重要的生存因子,切除該類神經(jīng)將經(jīng)由caspase-8介導(dǎo)的死亡受體通路誘發(fā)Leydig細(xì)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.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R346

【共引文獻(xiàn)】

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5 Albaha Barqawi,Herald Trummer,Randall Meacham;Effect of prolonged cryptorchidism on germ cell apoptosisand testicular sperm count[J];Asian Journal of Andrology;2004年01期

6 ;Involvement of nuclear factor-kappa B on corticosteroneinduced rat Leydig cell apoptosis[J];Asian Journal of Andrology;2006年06期

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本文編號(hào)：1916230