AQP1在睾丸缺血再灌注损伤中的表达

发布时间：2018-04-10 15:46

本文选题：睾丸 + 水通道蛋白1　；参考：《新疆医科大学》2014年硕士论文

【摘要】：目的：通过建立睾丸缺血再灌注损伤的模型,观察水通道蛋白1mRNA表达的变化与睾丸组织水肿程度的关系,探讨水通道蛋白1在睾丸缺血再灌注损伤中的作用。方法：采用睾丸动静脉结扎1h/再开通的方法,建立睾丸缺血再灌注损伤的模型。分别在术后12h,24h,36h,48h,5d取材,HE染色观察睾丸组织的病理改变,免疫组织化学法定位AQP1蛋白的表达,干湿比重法测量睾丸组织的含水量,荧光定量PCR法检测水通蛋白1mRNA的表达。结果：再灌注12h：术侧睾丸生精小管管间间隙增宽,生精上皮层数减少,管腔有少量精子,Johnsen评分8分；睾丸组织干湿比重值(85.67%±0.76%)高于假手术组(83.96%±0.45%)；AQP1基因的相对表达量为假手术组的1.94倍。再灌注24h：术侧睾丸生精小管内生精细胞大量脱落,生精上皮层状结构消失,管腔内几乎见不到精子,Johnsen评分6.5分；干湿比重值(87.33%±0.26%),达到了再灌注5d之内的最高值：AQP1基因的相对表达量为假手术组的4.95倍。再灌注36h、48h:术侧睾丸生精小管内各级生精细胞数量减少,排列紊乱,管腔中有精子生成,Johnsen评分7.5分；干湿比重值(85.98%±0.35%、85.13%±0.15%)与再灌注24h相比逐渐降低,但仍高于假手术组(83.96%±0.45%)；AQP1基因相对表达量与再灌注24h相比逐渐减少,分别为假手术组的3.20倍和2.43倍。再灌注5d：术侧睾丸生精小管管间间隙不再增宽,生精上皮内可见到大量未成熟的精子细胞,管腔中精子较少,Johnsen评分8.5分；干湿比重值(83.38%±0.15%)降低,且高于假手术组(83.96%±0.45%)；AQP1基因相对表达量为假手术组的1.33倍。以上结果差异均有统计学意义(P0.05)。非术侧睾丸在光镜下无明显病理改变,但12h、24h、36h、48h时干湿比重值随着术侧的变化有小幅度波动,与假手术组比较,差异有统计学意义(P0.05)；非术侧睾丸AQP1基因的表达亦有小幅度波动。免疫组织化学结果显示AQP1定位在睾丸血管内皮细胞膜及睾丸的结缔组织。结论：AQP1可能参与了睾丸缺血再灌注损伤的病理生理过程,AQP1基因表达量的变化可能对睾丸组织的水代谢有一定的影响。
[Abstract]:Objective: to establish the testicular ischemia reperfusion injury model, observe the relationship between the changes of water channel protein 1mRNA expression and testicular tissue edema, study of aquaporin 1 in testicular ischemia-reperfusion injury in rats. Methods: testicular arteriovenous ligation of 1h/ re opened, the establishment of testicular ischemia reperfusion injury model. After the operation. 12h, 24h, 36h, 48h, 5D materials, HE staining to observe the pathological changes of testis tissue, the expression of immunohistochemical localization of AQP1 protein, moisture content of testicular tissue dry wet method measurement, fluorescent quantitative detection of PCR expression of water through protein 1mRNA. Results: 12h of reperfusion: the operation side of seminiferous tubule tube gap widened, spermatogenicepithelium was reduced, the lumen of small sperm, Johnsen score of 8; testis wet proportion value (85.67% + 0.76%) was higher than that of sham group (83.96% + 0.45%); AQP1 Because of the relative expression quantity was 1.94 times of sham operation group. Reperfusion 24h: operative side of seminiferous tubule spermatogenic cell loss, seminiferous epithelium within the lumen of the lamellar structure disappeared, almost no sperm, Johnsen score of 6.5; the proportion of wet and dry value (87.33% + 0.26%), and reached the highest value 5D perfusion within: the relative expression of AQP1 gene was 4.95 times of the sham operation group. Reperfusion 36h, 48h: side of seminiferous tubule spermatogenic cells, reduce the number of disordered in the lumen of spermatogenesis, Johnsen score of 7.5; the proportion of wet and dry value (85.98% + 0.35%, 85.13% + 0.15%) compared with the reperfusion 24h gradually decreased, but still higher than the sham operation group (83.96% + 0.45%); the relative expression of AQP1 gene was compared with reperfusion 24h decreased gradually, were sham operated group 3.20 times and 2.43 times of reperfusion. 5D: operative side of seminiferous tubule pipe interval no longer wider, spermatogenic In the epithelium can be seen a large number of immature sperm cells, sperm in the lumen of small, Johnsen score of 8.5; the proportion of wet and dry value (83.38% + 0.15%) lower and higher than the sham operation group (83.96% + 0.45%); AQP1 gene relative expression quantity was 1.33 times of sham operation group. The differences were statistically significant (P0.05) non operative side testis. No obvious pathological changes, but 12h, 24h, 36h in the light microscope, 48h value with the change of dry wet operation side have small fluctuations, compared with the sham group, the difference was statistically significant (P0.05); the expression of AQP1 gene in non operative testes also have small fluctuations. The immunohistochemistry results showed that AQP1 was located in the testis and testicular vascular endothelial cell membrane of connective tissue. Conclusion: AQP1 may be involved in the pathophysiological process of testicular ischemia reperfusion injury, AQP1 gene expression changes of water metabolism on testicular tissue may have a certain Influence.

【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R697.22

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