本文關(guān)鍵詞： 睪丸扭轉(zhuǎn) c-kit PI3K 大鼠　出處：《遵義醫(yī)學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:了解大鼠性成熟前期一側(cè)睪丸扭轉(zhuǎn)后,對處于不同狀態(tài)的扭轉(zhuǎn)睪丸(扭轉(zhuǎn)未壞死睪丸、扭轉(zhuǎn)疑似壞死睪丸、扭轉(zhuǎn)壞死睪丸),特別是扭轉(zhuǎn)疑似壞死睪丸、扭轉(zhuǎn)壞死睪丸,采取何種手術(shù)方式(復(fù)位/切除),可最大限度降低性成熟期對側(cè)睪丸生精功能的損害。方法:(1)實驗方法:采用國際通用的Turner法制作性成熟前期的睪丸扭轉(zhuǎn)模型:56只3周齡雄性SD大鼠,隨機分成7組,每組8只,任選一組行假手術(shù),作為對照組(Control group,CG)。扭轉(zhuǎn)未壞死睪丸復(fù)位組(Non-necrotic testis detorsion group,NNTD):左側(cè)睪丸逆時針扭轉(zhuǎn)90°固定4h后復(fù)位睪丸;扭轉(zhuǎn)未壞死睪丸切除組(Non-necrotic testis orchiectomy group,NNTO):左側(cè)睪丸逆時針扭轉(zhuǎn)90°固定4h后切除睪丸。扭轉(zhuǎn)疑似壞死睪丸復(fù)位組(Suspected necrotic testis detorsion group,SNTD):左側(cè)睪丸逆時針扭轉(zhuǎn)360°固定4h后復(fù)位睪丸;扭轉(zhuǎn)疑似壞死睪丸切除組(Suspected necrotic testis orchiectomy group,SNTO):左側(cè)睪丸逆時針扭轉(zhuǎn)360°固定4h后切除睪丸。扭轉(zhuǎn)壞死睪丸復(fù)位組(Necrotic testis detorsion group,NTD):左側(cè)睪丸逆時針扭轉(zhuǎn)720°固定4h后復(fù)位睪丸;扭轉(zhuǎn)壞死睪丸切除組(Necrotic testis orchiectomy group,NTO):左側(cè)睪丸逆時針扭轉(zhuǎn)720°固定4h后切除睪丸。術(shù)后繼續(xù)飼養(yǎng)6周至性成熟期,采用快速脫頸椎法處死SD大鼠,切取對側(cè)睪丸石蠟包埋做光鏡下組織形態(tài)學(xué)觀察,以及免疫組織化學(xué)染色法檢測睪丸精原干細胞因子受體(Stem cell factor receptor,c-kit)及磷酯酰肌醇-3-激酶(Phosphoinositide 3-kinase,PI3K)的表達。同時抽取心臟血液離心后取血清,ELISA法檢測血清性激素水平(卵泡刺激素FSH、黃體生成素LH及睪酮T)。(2)統(tǒng)計學(xué)方法:實驗數(shù)據(jù)采用Excel進行整理,數(shù)據(jù)分析采用SPSS18.0完成。計量資料用`X±S形式表示,各組間指標差異的比較采用單因素分析進行,若差異具統(tǒng)計學(xué)意義,則采用lsd法進一步作兩兩比較。以p0.05為差異具統(tǒng)計學(xué)意義。結(jié)果:(1)組織形態(tài)學(xué)改變:cg組健側(cè)睪丸曲細精管內(nèi)生精上皮完好,無脫落,各級生精細胞均排列有序,無生精小管萎縮;nntd組和nnto組:健側(cè)睪丸均未出現(xiàn)明顯形態(tài)學(xué)改變;sntd組和snto組:輕微的形態(tài)學(xué)改變,主要表現(xiàn)為生精上皮細胞層數(shù)減少,少量生殖細胞脫落于管腔中;ntd組和nto組:前者睪丸生精上皮細胞層數(shù)減少明顯,部分生殖細胞脫落于管腔中,生殖細胞排列紊亂,可見生精小管萎縮,后者未出現(xiàn)明顯的形態(tài)學(xué)改變。(2)睪丸組織中生精細胞及間質(zhì)細胞的c-kit、pi3k的iod值結(jié)果:nntd組和nnto組間比較,差異無統(tǒng)計學(xué)意義(p0.05);sntd組和snto組比較,差異也無統(tǒng)計學(xué)意義(p0.05);但ntd組和nto組比較,ntd組中c-kit、pi3k的iod值均明顯低于nto(p0.05)。與假手術(shù)對照組比較,nntd組、nnto組及nto組中c-kit、pi3k的iod值均無明顯統(tǒng)計學(xué)差異(p0.05);與假手術(shù)對照組比較,sntd組、snto組以及ntd組中c-kit、pi3k的iod值均顯著降低(p0.05)。(3)血清性激素水平(卵泡刺激素fsh、黃體生成素lh及睪酮t):fsh、lh在各組中升高,t水平降低,但各性激素水平(fsh、lh、t)在nntd組與nnto組間比較、以及sntd組與snto組間比較,差異均無統(tǒng)計學(xué)意義(p0.05);性激素水平(lh、fsh、t)在ntd組與nto組間比較,ntd組的lh、fsh顯著升高,t明顯降低(p0.05)。cg組各性激素水平(fsh、lh、t)分別與nntd組、nnto組以及nto組間比較,差異均無統(tǒng)計學(xué)意義(p0.05);與cg組相比較,sntd組、snto組以及ntd組fsh、lh均升高,t均降低(p0.05)。結(jié)論:1、大鼠性成熟前期扭轉(zhuǎn)未壞死睪丸,無論復(fù)位還是切除,對性成熟期健側(cè)睪丸均無損傷;性成熟前期扭轉(zhuǎn)疑似壞死睪丸,無論復(fù)位還是切除,對性成熟期健側(cè)睪丸均有輕微損害;2、性成熟前期扭轉(zhuǎn)壞死睪丸的切除,對性成熟期健側(cè)睪丸具有保護作用,應(yīng)盡可能切除。
[Abstract]:Objective: To investigate the immature rats after unilateral testicular torsion, torsion of the testis in different condition (no necrosis of testis torsion, torsion of testicular torsion and necrosis of testis necrosis suspected, especially, suspected) torsion necrosis of testis, testicular torsion and necrosis, adopt surgical methods (reduction / removal), can reduce sexual maturity on the side of spermatogenic dysfunction. Methods: (1) experimental methods: immature using the Turner method of testicular torsion model: 56 3 week old male SD rats were randomly divided into 7 groups, 8 rats in each group, each group underwent sham operation as control group (Control group, CG). No necrosis of testis torsion reduction group (Non-necrotic testis detorsion group, NNTD): the left testicular torsion counter clockwise 90 DEG 4h after reduction of testicular torsion fixed; no necrosis testical resection group (Non-necrotic testis, orchiectomy group, NNTO): the left testis inverse 90 degrees clockwise twist fixed 4h after the resection of the testis. Testicular torsion suspected necrosis reduction group (Suspected necrotic testis detorsion group, SNTD): the left testicular torsion 360 degrees counterclockwise fixed 4H reset after testicular torsion mimicking necrosis; testical resection group (Suspected necrotic testis orchiectomy group, SNTO): the left testicular torsion 360 degrees counterclockwise fixed 4h after resection of the testis. Testicular torsion and necrosis (Necrotic testis detorsion group reduction group, NTD): the left testicular torsion 720 degrees counterclockwise fixed 4H reset after testicular torsion and necrosis; testicular resection group (Necrotic testis orchiectomy group, NTO): the left testicular torsion counter clockwise 720 DEG 4h after resection of fixed testes. After feeding for 6 weeks to continue sexual maturity, rapid removal of cervical SD rats were killed by cutting the contralateral testicular paraffin embedded for histological observation under light microscope, and immunohistochemical staining of testis Spermatogonial stem cell factor receptor (Stem cell factor receptor, c-kit) and phosphatidylinositol -3- kinase (Phosphoinositide, 3-kinase, PI3K) expression. At the same time from heart blood serum after centrifugation, serum sex hormone levels detected by ELISA (follicle stimulating hormone FSH, Huang Tisheng LH and testosterone T) (2). Statistical methods: experimental data using Excel collation, data analysis is completed by SPSS18.0. The measurement data is expressed by `X + S, was used to compare the differences among groups of single factor analysis, if the difference is statistically significant, LSD method was used to further compare with P0.05 22. The difference was statistically significant. Results: (1) change of tissue morphology: in group CG, the contralateral testis seminiferous tubules seminiferous epithelium intact, no shedding, spermatogenic cells were arranged in order, no seminiferous tubule atrophy; nntd group and nnto group: the contralateral testis showed no obvious morphological change Variable; sntd group and SNTO group: slight alteration in morphology, mainly for the seminiferous epithelial cell layers decreased, a small amount of germ cells shed from the lumen; NTD group and nto group: the seminiferous epithelial cell layers decreased obviously, part of germ cell shedding in the lumen, germ cell disorder, visible seminiferous tubule the latter did not shrink, there were obvious changes. (2) in the testis spermatogenic cells and interstitial cells of c-kit, PI3K IOD results: nntd group and nnto group comparison, no statistically significant difference (P0.05); sntd group and SNTO group, there was no significant difference (P0.05); but the NTD group and NTO group, c-kit NTD group, PI3K IOD was obviously lower than that of NTO (P0.05). The control group compared with the sham nntd group, c-kit nnto group and nto group, there were no statistically significant differences in PI3K values of IOD (P0.05); the control group compared with sham operation sntd group, SNTO group 鍙妌td緇勪腑c-kit,pi3k鐨刬od鍊煎潎鏄捐憲闄嶄綆(p0.05).(3)琛，

本文編號：1468129