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AQP1在睪丸缺血再灌注損傷中的表達(dá)

發(fā)布時(shí)間:2018-04-10 15:46

  本文選題:睪丸 + 水通道蛋白1; 參考:《新疆醫(yī)科大學(xué)》2014年碩士論文


【摘要】:目的:通過建立睪丸缺血再灌注損傷的模型,觀察水通道蛋白1mRNA表達(dá)的變化與睪丸組織水腫程度的關(guān)系,探討水通道蛋白1在睪丸缺血再灌注損傷中的作用。方法:采用睪丸動(dòng)靜脈結(jié)扎1h/再開通的方法,建立睪丸缺血再灌注損傷的模型。分別在術(shù)后12h,24h,36h,48h,5d取材,HE染色觀察睪丸組織的病理改變,免疫組織化學(xué)法定位AQP1蛋白的表達(dá),干濕比重法測量睪丸組織的含水量,熒光定量PCR法檢測水通蛋白1mRNA的表達(dá)。結(jié)果:再灌注12h:術(shù)側(cè)睪丸生精小管管間間隙增寬,生精上皮層數(shù)減少,管腔有少量精子,Johnsen評分8分;睪丸組織干濕比重值(85.67%±0.76%)高于假手術(shù)組(83.96%±0.45%);AQP1基因的相對表達(dá)量為假手術(shù)組的1.94倍。再灌注24h:術(shù)側(cè)睪丸生精小管內(nèi)生精細(xì)胞大量脫落,生精上皮層狀結(jié)構(gòu)消失,管腔內(nèi)幾乎見不到精子,Johnsen評分6.5分;干濕比重值(87.33%±0.26%),達(dá)到了再灌注5d之內(nèi)的最高值:AQP1基因的相對表達(dá)量為假手術(shù)組的4.95倍。再灌注36h、48h:術(shù)側(cè)睪丸生精小管內(nèi)各級(jí)生精細(xì)胞數(shù)量減少,排列紊亂,管腔中有精子生成,Johnsen評分7.5分;干濕比重值(85.98%±0.35%、85.13%±0.15%)與再灌注24h相比逐漸降低,但仍高于假手術(shù)組(83.96%±0.45%);AQP1基因相對表達(dá)量與再灌注24h相比逐漸減少,分別為假手術(shù)組的3.20倍和2.43倍。再灌注5d:術(shù)側(cè)睪丸生精小管管間間隙不再增寬,生精上皮內(nèi)可見到大量未成熟的精子細(xì)胞,管腔中精子較少,Johnsen評分8.5分;干濕比重值(83.38%±0.15%)降低,且高于假手術(shù)組(83.96%±0.45%);AQP1基因相對表達(dá)量為假手術(shù)組的1.33倍。以上結(jié)果差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。非術(shù)側(cè)睪丸在光鏡下無明顯病理改變,但12h、24h、36h、48h時(shí)干濕比重值隨著術(shù)側(cè)的變化有小幅度波動(dòng),與假手術(shù)組比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05);非術(shù)側(cè)睪丸AQP1基因的表達(dá)亦有小幅度波動(dòng)。免疫組織化學(xué)結(jié)果顯示AQP1定位在睪丸血管內(nèi)皮細(xì)胞膜及睪丸的結(jié)締組織。結(jié)論:AQP1可能參與了睪丸缺血再灌注損傷的病理生理過程,AQP1基因表達(dá)量的變化可能對睪丸組織的水代謝有一定的影響。
[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.

【學(xué)位授予單位】:新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2014
【分類號(hào)】:R697.22

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