大鼠性成熟前期睾丸扭转对性成熟期健侧睾丸c-kit、PI3K表达的影响

发布时间：2018-01-27 09:20

  本文关键词： 睾丸扭转 c-kit PI3K 大鼠　出处：《遵义医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:了解大鼠性成熟前期一侧睾丸扭转后,对处于不同状态的扭转睾丸(扭转未坏死睾丸、扭转疑似坏死睾丸、扭转坏死睾丸),特别是扭转疑似坏死睾丸、扭转坏死睾丸,采取何种手术方式(复位/切除),可最大限度降低性成熟期对侧睾丸生精功能的损害。方法:(1)实验方法:采用国际通用的Turner法制作性成熟前期的睾丸扭转模型:56只3周龄雄性SD大鼠,随机分成7组,每组8只,任选一组行假手术,作为对照组(Control group,CG)。扭转未坏死睾丸复位组(Non-necrotic testis detorsion group,NNTD):左侧睾丸逆时针扭转90°固定4h后复位睾丸;扭转未坏死睾丸切除组(Non-necrotic testis orchiectomy group,NNTO):左侧睾丸逆时针扭转90°固定4h后切除睾丸。扭转疑似坏死睾丸复位组(Suspected necrotic testis detorsion group,SNTD):左侧睾丸逆时针扭转360°固定4h后复位睾丸;扭转疑似坏死睾丸切除组(Suspected necrotic testis orchiectomy group,SNTO):左侧睾丸逆时针扭转360°固定4h后切除睾丸。扭转坏死睾丸复位组(Necrotic testis detorsion group,NTD):左侧睾丸逆时针扭转720°固定4h后复位睾丸;扭转坏死睾丸切除组(Necrotic testis orchiectomy group,NTO):左侧睾丸逆时针扭转720°固定4h后切除睾丸。术后继续饲养6周至性成熟期,采用快速脱颈椎法处死SD大鼠,切取对侧睾丸石蜡包埋做光镜下组织形态学观察,以及免疫组织化学染色法检测睾丸精原干细胞因子受体(Stem cell factor receptor,c-kit)及磷酯酰肌醇-3-激酶(Phosphoinositide 3-kinase,PI3K)的表达。同时抽取心脏血液离心后取血清,ELISA法检测血清性激素水平(卵泡刺激素FSH、黄体生成素LH及睾酮T)。(2)统计学方法:实验数据采用Excel进行整理,数据分析采用SPSS18.0完成。计量资料用`X±S形式表示,各组间指标差异的比较采用单因素分析进行,若差异具统计学意义,则采用lsd法进一步作两两比较。以p0.05为差异具统计学意义。结果:(1)组织形态学改变:cg组健侧睾丸曲细精管内生精上皮完好,无脱落,各级生精细胞均排列有序,无生精小管萎缩;nntd组和nnto组:健侧睾丸均未出现明显形态学改变;sntd组和snto组:轻微的形态学改变,主要表现为生精上皮细胞层数减少,少量生殖细胞脱落于管腔中;ntd组和nto组:前者睾丸生精上皮细胞层数减少明显,部分生殖细胞脱落于管腔中,生殖细胞排列紊乱,可见生精小管萎缩,后者未出现明显的形态学改变。(2)睾丸组织中生精细胞及间质细胞的c-kit、pi3k的iod值结果:nntd组和nnto组间比较,差异无统计学意义(p0.05);sntd组和snto组比较,差异也无统计学意义(p0.05);但ntd组和nto组比较,ntd组中c-kit、pi3k的iod值均明显低于nto(p0.05)。与假手术对照组比较,nntd组、nnto组及nto组中c-kit、pi3k的iod值均无明显统计学差异(p0.05);与假手术对照组比较,sntd组、snto组以及ntd组中c-kit、pi3k的iod值均显著降低(p0.05)。(3)血清性激素水平(卵泡刺激素fsh、黄体生成素lh及睾酮t):fsh、lh在各组中升高,t水平降低,但各性激素水平(fsh、lh、t)在nntd组与nnto组间比较、以及sntd组与snto组间比较,差异均无统计学意义(p0.05);性激素水平(lh、fsh、t)在ntd组与nto组间比较,ntd组的lh、fsh显著升高,t明显降低(p0.05)。cg组各性激素水平(fsh、lh、t)分别与nntd组、nnto组以及nto组间比较,差异均无统计学意义(p0.05);与cg组相比较,sntd组、snto组以及ntd组fsh、lh均升高,t均降低(p0.05)。结论:1、大鼠性成熟前期扭转未坏死睾丸,无论复位还是切除,对性成熟期健侧睾丸均无损伤;性成熟前期扭转疑似坏死睾丸,无论复位还是切除,对性成熟期健侧睾丸均有轻微损害;2、性成熟前期扭转坏死睾丸的切除,对性成熟期健侧睾丸具有保护作用,应尽可能切除。
[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缁勪腑c-kit,pi3k鐨刬od鍊煎潎鏄捐憲闄嶄綆(p0.05).(3)琛，

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