雌激素介导的Aquaporin2在女性压力性尿失禁患者尿道支撑组织成纤维细胞上的表达和意义

发布时间：2018-05-27 10:45

本文选题：压力性尿失禁 + AQP2　；参考：《浙江大学》2014年硕士论文

【摘要】：背景和目的：压力性尿失禁(stress urinary incontinence, SUI)是中老年妇女常见慢性病,是指在腹压增加时,出现不自主的尿液自尿道外口溢出。SUI的病因很复杂,年龄是其发病的主要诱因之一,但是发病机制迄今未明,最经典的“整体理论”和“吊床假说”均强调尿道支撑结构薄弱导致SUI的发生。尿道支撑结构由大量细胞外基质(extracellular matrix, ECM)和少量成纤维细胞等组成。有研究表明ECM重构障碍是SUI发病的关键因素,而雌激素直接参与了ECM的重构过程。水通道蛋白(Aquaporins AQPs)是选择性高效转运水分子的通道,同时还广泛参与人体细胞的迁移运动,与多种组织的损伤修复有关,而且有研究发现在水通道蛋白2(Aquaporin2,AQP2)的基因上存在雌激素的反应元件,证实了雌激素能直接调控AQP2的基因转录而调节其在子宫内膜细胞上的表达,从而促进内膜细胞的迁移、入侵、粘附及增殖。本研究拟通过原代培养获得女性尿道支撑组织成纤维细胞,采用免疫荧光技术证实AQP2在尿道支撑组织成纤维细胞上的表达,采用Western blot法观测绝经前后SUI患者与非SUI患者尿道支撑组织成纤维细胞上AQP2的表达差异,探讨AQP2在SUI发生发展中的作用；同时,通过测定血清雌二醇(estradiol,E2)水平,探讨尿道支撑组织成纤维细胞上AQP2的表达水平与血清E2水平的相关性。方法：收集浙江大学医学院附属妇产科医院2013年1月至2014年3月收治的18例SUI患者(SUI组)和18例非压力性尿失禁妇女(对照组)的阴道前壁组织(即尿道支撑组织),每组再分为绝经前后两组各9例,所有组织标本来源于SUI患者行抗尿失禁手术时和非SUI患者因尿道旁腺囊肿、尿道憩室、尿道附近阴道壁囊肿等疾病手术时,取材于尿道附近约0.5×1cm2的阴道前壁全层组织。取组织块进行成纤维细胞原代培养,并予免疫组化鉴定。采用免疫荧光法测定成纤维细胞上AQPs各亚型的表达,采用Western blot法检测成纤维细胞上AQP2蛋白表达水平,比较绝经前后SUI组和非SUI组之间尿道支撑组织成纤维细胞上AQP2蛋白表达水平的差异。所有36例入选对象均在外科手术前一天早晨7时空腹采静脉血2ml,采用电化学发光免疫检测(ECLIA)法检测血清E2水平,采用Pearson相关性分析法探讨尿道支撑组织成纤维细胞上AQP2蛋白表达水平与血清E2水平之间的相关性。结果： 1.免疫荧光技术和Western blot法均发现女性尿道支撑组织成纤维细胞上有AQP2的表达； 2.经t检验,绝经前后SUI组和对照组的年龄、分娩次数、体重指数以及绝经后SUI组和对照组的绝经年限差异均无统计学意义；绝经前血清E2水平SUI组(311.43±93.63nmol/L)显著低于对照组(530.30±232.21nmol/L)(P0.05).绝经后血清E2水平SUI组为39.91±16.76nmol/L,对照组为31.30±9.95nmol/L两组之间差异无统计学意义(P0.05)；经相关性分析,血清E2水平与所有标本尿道支撑组织成纤维细胞(r=—0.537,P0.05)及总的SUI组标本尿道支撑组织成纤维细胞(r=—0.860,P0.05)上AQP2的表达水平均存在显著的负相关,而与总的对照组标本尿道支撑组织成纤维细胞上AQP2的表达水平无显著相关(P0.05)。 3.经Western blot法检测,绝经前后SUI组及绝经前后对照组尿道支撑组织成纤维细胞上均有AQP2的表达,总的SUI组和总的对照组尿道支撑组织成纤维细胞上AQP2的蛋白表达水平分别为26.29±2.53和25.32±3.80,t检验显示两组间差异无统计学意义(P0.05)；总的绝经前组和总的绝经后组尿道支撑组织成纤维细胞上AQP2的蛋白表达水平分别为23.25±2.40和28.36±1.31,经t检验,总的绝经后组尿道支撑组织成纤维细胞上AQP2的表达明显升高(P0.05)绝经前SUI组和绝经后SUI组尿道支撑组织成纤维细胞上AQP2的蛋白表达水平分别为24.03±0.98和28.54±1.11,绝经前对照组和绝经后对照组尿道支撑组织成纤维细胞上AQP2的蛋白表达水平分别为22.46±3.15和28.19+1.54,经t检验,绝经前SUI组与绝经前对照组、绝经后SUI组与绝经后对照组上的表达差异均无统计学意义(P均0.05)；绝经后SUI组尿道支撑组织成纤维细胞上AQP2的表达较绝经前SUI组上的表达明显升高,两组间的比较差异有统计学意义(P0.05)；绝经后对照组尿道支撑组织成纤维细胞上AQP2的表达较绝经前对照组上的表达亦明显升高,两组间的比较差异有统计学意义(P0.05)。结论： 1.女性尿道支撑组织成纤维细胞上存在AQP2的表达； 2.AQP2的表达与雌激素水平密切相关,绝经后SUI组尿道支撑组织成纤维细胞上AQP2的表达较绝经前SUI组上的表达明显升高,可能是绝经后尿道支撑组织成纤维细胞上低雌激素水平上调AQP2的表达所致。
[Abstract]:Background and Purpose :

Aquaporins ( AQPs ) is a key factor for selective high - efficiency transport of water molecules , and it has been found that estrogen can regulate the expression of AQP2 directly and regulate the expression of AQP2 on endometrial cells .
At the same time , the level of serum estradiol ( E2 ) was measured , and the correlation between AQP2 expression level and serum E2 level was investigated .

Method :

The expression of AQP2 protein in fibroblasts from January 2013 to March 2014 was collected from January 2013 to March 2014 in the Affiliated Hospital of Zhejiang University Medical College from January 2013 to March 2014 .

The serum levels of E2 were measured by electrochemical luminescence immunoassay ( ECLIA ) and Pearson correlation analysis was used to investigate the correlation between AQP2 protein expression level and serum E2 level .

Results :

1 . The expression of AQP2 was found in the fibroblasts of female urethra by immunofluorescence technique and Western blot .

2 . After t test , there was no significant difference between the age , the number of deliveries , the body mass index and the menopausal period in the control group before and after menopause .
The serum level of E2 in the postmenopausal serum was significantly lower than that of the control group ( 530.30 卤 232.21nmol / L ) ( P0.05 ) . The serum level of E2 in postmenopausal serum was 39.91 卤 16.76nmol / L and 31.30 卤 9.95nmol / L in the control group ( P0.05 ) .
The level of serum E2 was negatively correlated with the expression level of AQP2 in all specimens of urethral support tissue fibroblasts ( r = - 0.537 , P0.05 ) , and the expression level of AQP2 was not significantly correlated with the total control group ( P0.05 ) .

3 . The expression level of AQP2 was detected by Western blot , and the expression level of AQP2 was 26.29 卤 2.53 and 25.32 卤 3.80 respectively , and there was no significant difference between the two groups ( P0.05 ) .
The expression levels of AQP2 were 23.25 卤 2.40 and 28.36 卤 1.31 , respectively , and the protein expression levels of AQP2 were 24.03 卤 0.98 and 28.54 卤 1.11 respectively .
The expression of AQP2 was significantly higher than that in premenopausal women , but there was significant difference between the two groups ( P0.05 ) .
Compared with the control group , the expression of AQP2 in the control group was significantly higher than that in the control group ( P0.05 ) .

Conclusion :

1 . The expression of AQP2 was found in the fibroblasts of female urethra .

2 . The expression of AQP2 is closely related to the level of estrogen , and the expression of AQP2 is obviously higher than that in premenopausal women , which may be the result of the up - regulation of AQP2 by low estrogen level in the post - menopausal urethral support tissue fibroblasts .
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R711.59

【相似文献】