【摘要】：背景： 盆腔臟器脫垂（Pelvic Organ Prolapse, POP）是中老年婦女中的常見婦科疾病，對患者的健康和生活質(zhì)量有著嚴(yán)重影響，隨著社會老齡化進程的加快，POP受到的關(guān)注與日俱增。 POP的病因復(fù)雜，涉及多個基因的改變和多種mRNA及其蛋白質(zhì)的表達。本實驗研究了LOXL1基因單核苷酸多態(tài)性（single nucleotide polymorphism,SNPs）以及β-catenin mRNA的表達與POP的關(guān)系，初步探討了它們在POP發(fā)病機制中的作用。 目的： 研究LOXL1基因SNPs以及β-catenin mRNA的表達與POP發(fā)生、發(fā)展之間的關(guān)系，從分子水平進一步解釋盆腔臟器脫垂的病因?qū)W機制。 方法： 選擇盆腔臟器脫垂患者29例，同時選擇無POP的良性婦科疾病患者29例作為對照組。從全血中提取基因組DNA，對LOXL1基因片段進行擴增，對擴增產(chǎn)物進行直接測序，分析rs1048661位點及rs3825942位點的基因多態(tài)性。 選擇因盆腔臟器脫垂行全子宮切除術(shù)的患者17例，同時選擇無POP的、因良性婦科疾病行全子宮切除術(shù)的患者17例作為對照組。取主韌帶組織2塊，1塊用于提取RNA，使用RT-PCR技術(shù)獲得β-catenin的cDNA,將產(chǎn)物進行電泳并計算相對吸光度。另1塊制成組織切片，使用麗春紅-苦味酸-維多利亞藍染色法進行染色，觀察其中膠原纖維及彈性纖維的含量。 結(jié)果： 1、rs1048661位點SNPs，盆腔臟器脫垂患者的等位基因構(gòu)成比、基因型構(gòu)成比與對照組無統(tǒng)計學(xué)差異（P值均0.05）。在rs3825942位點，盆腔臟器脫垂組等位基因G的構(gòu)成比顯著高于對照組，，P=0.029，兩組間差異具有統(tǒng)計學(xué)意義。無論是盆腔臟器脫垂患者組還是對照組，均未在rs3825942位點發(fā)現(xiàn)AA基因型。盆腔臟器脫垂患者GG基因型頻率顯著高于對照組，P=0.021。 2、盆腔臟器脫垂患者主韌帶中β-catenin mRNA含量顯著低于對照組，P=0.02。 3、盆腔臟器脫垂患者子宮主韌帶組織中膠原纖維及彈性纖維相對吸光度值明顯均低于對照組（P=0.010，P=0.006），差異有統(tǒng)計學(xué)意義。β-catenin mRNA在子宮主韌帶中的表達量與子宮主韌帶中的膠原纖維含量之間存在直線回歸關(guān)系，P 0.01；而β-catenin mRNA的表達量與彈性纖維的含量之間，不存在直線回歸關(guān)系，P0.05。 結(jié)論： 1、LOXL1的單核苷酸多態(tài)性與中國漢族女性盆腔臟器脫垂的發(fā)病有關(guān)。 2、 rs3825942單核苷酸多態(tài)性位點，等位基因G是盆腔臟器脫垂的危險等位基因，GG基因型是盆腔臟器脫垂的危險基因型。 3、盆腔臟器脫垂組患者子宮主韌帶組織中β-catenin mRNA表達量下降。 4、盆腔臟器脫垂組患者主韌帶中的彈性纖維和膠原纖維含量明顯降低。 5、子宮主韌帶中β-catenin mRNA表達量降低與膠原纖維含量降低呈直線相關(guān)關(guān)系。
[Abstract]:Background: pelvic viscera prolapse (Pelvic Organ Prolapse, POP) is a common gynecological disease in middle-aged and elderly women, which has a serious impact on patients' health and quality of life. With the acceleration of the aging process of society, POP is receiving increasing attention. The etiology of POP is complex, involving multiple gene changes and the expression of multiple mRNA and its proteins. In this study, we studied the relationship between LOXL1 gene single nucleotide polymorphisms (single nucleotide polymorphism,SNPs), 尾-catenin mRNA expression and POP, and discussed their roles in the pathogenesis of POP. Objective: to study the relationship between the expression of SNPs and 尾-catenin mRNA of LOXL1 gene and the occurrence and development of POP, and to further explain the etiological mechanism of pelvic organ prolapse at molecular level. Methods: 29 patients with pelvic organ prolapse and 29 patients with benign gynecological diseases without POP were selected as control group. Genomic DNA, was extracted from whole blood to amplify the LOXL1 gene fragment. The amplified product was sequenced directly and the polymorphism of rs1048661 locus and rs3825942 locus was analyzed. Seventeen patients with pelvic organ prolapse underwent total hysterectomy, 17 patients without POP and 17 patients with benign gynecologic diseases were selected as control group. The main ligaments were extracted from two pieces of tissue and one piece was used to extract RNA,. The 尾-catenin cDNA, was obtained by RT-PCR technique. The product was electrophoretic and the relative absorbance was calculated. The other one was made into tissue section and stained with Victorian blue staining method. The contents of collagen and elastic fibers were observed. Results: 1 the allelic and genotypic ratios of patients with SNPs, pelvic organ prolapse at rs1048661 were not significantly different from those of the control group (P < 0. 05). At the rs3825942 locus, the G ratio of allele G in pelvic organ prolapse group was significantly higher than that in control group, and the difference between the two groups was statistically significant. Neither the pelvic organ prolapse group nor the control group found AA genotypes at rs3825942 locus. The frequency of GG genotype in patients with pelvic organ prolapse was significantly higher than that in control group (P < 0.021). 2, the content of 尾-catenin mRNA in the main ligament of pelvic viscera prolapse patients was significantly lower than that in the control group (P < 0. 02). 3. The relative absorbance of collagen fiber and elastic fiber in the main ligament of uterus in patients with prolapse of pelvic viscera was significantly lower than that in the control group (P0. 010, P0. 006). There was a linear regression relationship between the expression of 尾-catenin mRNA in the main uterine ligament and the content of collagen fiber in the main uterine ligament (P 0.01). However, there was no linear regression relationship between the expression of 尾-catenin mRNA and the content of elastic fiber, P0.05. Conclusion: 1 the single nucleotide polymorphism of LOXL1 is associated with pelvic organ prolapse in Chinese Han women. 2, rs3825942 single nucleotide polymorphism locus, allele G is the dangerous allele of pelvic organ prolapse, GG genotype is the dangerous genotype of pelvic organ prolapse. 3. The expression of 尾-catenin mRNA in the main ligament of uterus decreased in pelvic organ prolapse group. 4. The content of elastic fiber and collagen fiber in the main ligament of pelvic viscera prolapse group was significantly decreased. 5. There was a linear correlation between the decrease of 尾-catenin mRNA expression and the decrease of collagen fiber content in the main ligament of uterus.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R711.2

【參考文獻】

中國期刊全文數(shù)據(jù)庫 前4條

1 張琳琳;于浩;張師前;;絕經(jīng)前盆腔器官膨出患者子宮主韌帶中Elastin、LOX基因表達的研究[J];山東大學(xué)學(xué)報(醫(yī)學(xué)版);2008年02期

2 龍素萍;張振武;李義星;;MMP-3和CollagenⅢ在盆底器官脫垂患者陰道壁組織中表達的研究[J];中國醫(yī)藥指南;2008年19期

3 任琛琛;;POP患者盆底組織中MMP-3、TIMP-3的研究[J];中國婦幼保健;2008年20期

4 朱蘭,郎景和,馮瑞娥,陳杰;絕經(jīng)后壓力性尿失禁患者盆底支持結(jié)構(gòu)雌激素受體的研究[J];中華婦產(chǎn)科雜志;2004年10期

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