性激素受体在良性前列腺增生症中的发病机制研究

发布时间：2018-05-07 10:10

本文选题：良性前列腺增生症 + 微小RNA　；参考：《第三军医大学》2016年博士论文

【摘要】：研究背景:良性前列腺增生症(benign prostatic hyperplasia,BPH)是一种常见于老年男性的由前列腺移行区上皮和基质良性增生造成膀胱出口梗阻,从而引起患者一系列下尿路症状的良性疾病,其发病率与年龄呈正相关。目前我国即将进入老龄化社会,由BPH带来的卫生经济学压力更加严峻。目前认为BPH是一种由多因素参与的具有复杂发病机制的疾病,到目前为止其确切发病机制及病因仍不清楚。于是明确BPH的确切发病机制将有可能发现更加有效的预防及治疗方案,对提高老年男性的生活质量、减少社会医疗保障负担具有重要的意义。研究目的:本研究针对性激素受体包括雄激素受体(AR)、雌激素受体α(ERα)、雌激素受体β(ERβ)和孕激素受体(PGR)在BPH中的可能机制做一探索性研究,期望能找到一个突破口。鉴于BPH的可能发病机制的多因素性,在明确上述性激素受体在BPH组织中的确切表达变化后,拟从可能参与性激素受体m RNA调控的micro RNAs、可能受性激素受体调控且与细胞增殖或凋亡相关的细胞因子和生长因子、代谢综合征与性激素受体的相关性等多方面进行探索,在研究过程中根据实验发现结果及时调整方案,为进一步深入研究并揭示性激素受体在BPH中的确切发病机制提供一定的思路。研究方法:1.收集2015年2月~2015年6月在第三军医大学第一附属医院泌尿外科住院并行手术治疗且病例资料完整的BPH病例40例,同时收集器官捐献者以及其他手术切除前列腺并通过病理诊断明确为正常前列腺的病例5例。2.通过联合丙酸睾酮和苯甲酸雌二醇诱导去势SD雄性大鼠建立大鼠BPH模型。3.联合使用IHC、qRT-PCR和WB的方法确定性激素受体包括AR、ERα、ERβ和PGR在BPH组织中较正常前列腺组织的表达差异,然后采用同样方法检测大鼠BPH模型组织中这些受体的表达差异变化。4.通过芯片检测初步了解及筛查BPH组织中mi RNAs是否存在异常表达,然后通过qRT-PCR的方法对mi RNAs表达芯片提示可能具有异常表达的mi RNAs的确切表达进行验证。5.采用qRT-PCR的方法检测BPH组织中可能受性激素受体调控且有可能与BPH发病相关的细胞因子、生长因子在转录水平的表达差异情况。6.根据MetS的诊断标准对纳入研究的BPH病例进行分组,分为BPH伴MetS组和BPH不伴MetS组,并对比分析两组间临床资料差异性明确MetS与BPH的相关性,然后使用qRT-PCR及WB的研究方法明确两组各性激素受体的表达差异情况。研究结果:1.在人BPH组织中AR和PGR表达显著升高,ERα的表达显著下降;大鼠BPH模型实验组中PGR的表达显著上调,AR和ERα的表达显著下调;ERβ的表达未见显著性差异;IHC、qRT-PCR、WB在检测上述性激素受体表达的结果一致。2.mi RNAs芯片检测结果发现发现mi R-126-3p和mi R-4672在BPH组织中表达下调具有统计学意义,但是qRT-PCR的验证结果未见mi R-126-3p、mi R-4672、mi R-143-3p和mi R-145-3p在BPH组织中和正常前列腺组织中的表达具有显著性差异。3.采用qRT-PCR方法检测发现ETV1、TLR1基因转录水平的表达在BPH组织中显著性上调,而TGFβ1-SMAD2/3、IL6R-JAK2-STAT3、KRAS-ERK、TLR2基因的转录水平表达在BPH组织和正常前列腺组织中的差异无统计学意义。4.BPH不伴MetS组与BPH伴MetS组的临床资料分析:两组之间年龄相仿;在代谢综合征相关各诊断因素上,如BMI、空腹血糖、甘油三酯、高密度脂蛋白、血压,两组间具有显著性差异(P0.05);在BPH相关的检查检验结果方面,国际前列腺症状评分以及最大尿流率在两组间的差异并无统计学意义(P0.05),但总前列腺特异性抗原和前列腺体积在BPH伴MetS组显著性升高(P0.05);组织炎症情况对比方面,BPH不伴MetS组内的患者前列腺组织炎症检出率为46.7%(7/15),而BPH伴MetS组的患者前列腺组织炎症检出率为76.2%(16/21),但该差异无统计学意义(P0.05);采用qRT-PCR和WB法的检测发现在伴有MetS的BPH患者的前列腺组织中AR、ERα和PGR在转录水平和蛋白水平的表达均显著性升高(P0.05),而ERβ的表达在两组间未见显著性差异(P0.05)。研究结论:1.证实了BPH组织中AR和PGR的表达上调以及ERα的表达下调。推测在BPH中AR和PGR具有诱导疾病起病、促进疾病进展的作用,而ERα则具有抑制作用。激素诱导大鼠BPH模型与人BPH中雄激素/AR通路的活化机制具有差异性。2.就目前该研究的数据来看,尚不能认为mi RNAs与BPH组织中性激素受体的异常表达具有相关性,也不能认为mi RNAs在BPH发病机制中具有显著的作用。在筛查BPH组织中mi RNAs差异表达方面需要灵敏度和特异度更高的实验方法,同时需要扩大正常前列腺样本量继续验证以排除样本偏倚的影响。3.通过目前所得到的实验数据,发现BPH组织中ETV1、TLR1基因在转录水平的表达较正常前列腺组织显著性上调,而其余所检测的因子在转录水平表达则无统计学意义。但是尚不能就此实验结果予以下结论,还需要进一步研究明确这些因子蛋白水平及蛋白磷酸化水平的变化。初步推测AR-ETV1通路可能与BPH的发病具有相关性,也是我们深入研究的可能方向。4.证实了MetS和BPH的相关性,伴有MetS的BPH的患者前列腺总体积更大、血清PSA水平更高、组织炎症更显著,但目前尚不能认为MetS对BPH的相关症状有显著性的影响。发现伴MetS的BPH患者前列腺组织内AR、ERα和PGR表达显著性上调。推测MetS可诱导前列腺组织AR、ERα和PGR表达上调,促进BPH的发生及疾病进展。
[Abstract]:Background: benign prostatic hyperplasia (BPH) is a kind of benign disease common in elderly men, which is caused by epithelial and benign hyperplasia of the prostatic migrating area, causing a series of benign diseases of lower urinary tract symptoms. The incidence of the disease is positively related to age. The pressure of health economics brought by BPH is more severe. BPH is now considered to be a complex disease with multiple factors, and the exact pathogenesis and cause of the disease are still unclear so far. Therefore, it is clear that the exact pathogenesis of BPH will be possible to find more effective prevention and treatment programs and to improve the old. The quality of life of men is of great significance in reducing the burden of social health care. Objective: This study aims to explore the mechanism of the ability of sex hormone receptors, including androgen receptor (AR), estrogen receptor alpha (ER alpha), estrogen receptor beta (ER beta) and progestin receptor (PGR) in BPH, to find a breakthrough. The multiple factors of the possible pathogenesis of BPH, after identifying the exact changes in the expression of the sex hormone receptor in the BPH tissue, are intended to be regulated by the micro RNAs, which may be regulated by the sex hormone receptor m RNA, and may be regulated by the hormone receptor and the cytokines and growth factors associated with cell proliferation or apoptosis, metabolic syndrome and sex hormone receptor In order to further study and reveal the exact pathogenesis of sex hormone receptor in BPH, the research method: 1. collected in the Department of Urology, the First Affiliated Hospital of Third Military Medical University, February 2015, and in the Department of Urology of the First Affiliated Hospital of Third Military Medical University. 40 cases of BPH cases with complete surgical treatment and case data were collected and 5 cases of.2. were combined with testosterone propionate and estradiol benzoate to establish rat BPH model.3. combined with IHC, qRT-PCR and WB using IHC, qRT-PCR and WB. Methods deterministic hormone receptors, including AR, ER a, ER beta and PGR, were different in the expression of normal prostate tissue in BPH tissues. Then the same method was used to detect the differences in the expression of these receptors in the rat BPH model tissue..4. was detected by chip detection and screened for the existence of abnormal expression of MI RNAs in BPH tissues and then through qRT-P. CR's approach to the MI RNAs expression chip suggesting the exact expression of MI RNAs that may have abnormal expression..5. uses qRT-PCR to detect cytokines in BPH tissues that may be regulated by the sex hormone receptor and may be associated with the pathogenesis of BPH. The difference of the growth factor at the transcriptional level of.6. is based on MetS diagnostic criteria The BPH cases included in the study were divided into groups of BPH with MetS and BPH without MetS, and the correlation between MetS and BPH was clearly defined in the two groups. Then the expression difference between the two groups of sex hormone receptors was determined by qRT-PCR and WB. The results were as follows: 1. the AR and PGR expressions were significantly increased in the human BPH tissue. The expression of ER alpha was significantly decreased, and the expression of PGR in the experimental group of BPH was significantly up-regulated, and the expression of AR and ER alpha was significantly down, and there was no significant difference in the expression of ER beta. IHC, qRT-PCR, and WB detected the expression of the above sex hormone receptors by.2.mi RNAs chips. The down regulation was statistically significant, but the results of qRT-PCR's verification were not mi R-126-3p, MI R-4672, MI R-143-3p and MI R-145-3p in the BPH tissues and normal prostate tissues. The transcriptional levels of IL6R-JAK2-STAT3, KRAS-ERK, and TLR2 genes were expressed in BPH tissues and normal prostate tissues with no statistical significance,.4.BPH was not associated with the clinical data of the MetS group and the BPH with the MetS group: the age of the two groups was similar; in the metabolic syndrome related diagnostic factors, such as BMI, fasting blood glucose, triglycerides, and high-density lipoprotein There was a significant difference between the two groups (P0.05). In the BPH related examination results, the International Prostatic Symptom Score and the maximum urinary flow rate were not statistically significant between the two groups (P0.05), but the total prostatic specific antigen and the prostate volume were significantly higher in the BPH group (P0.05), and the tissue inflammation was compared. The positive rate of prostatic tissue inflammation in the patients with BPH without MetS was 46.7% (7/15), while the rate of prostatic tissue inflammation in the BPH and MetS group was 76.2% (16/21), but the difference was not statistically significant (P0.05). The detection of the prostate tissue with qRT-PCR and WB was found in the prostate tissue of BPH patients with MetS and at the transcriptional level and at the transcriptional level. The expression of protein level was significantly increased (P0.05), but the expression of ER beta was not significantly different between the two groups (P0.05). 1. confirmed the up regulation of AR and PGR expression in BPH tissue and the downregulation of ER a. It is suggested that AR and PGR have the effect of inducing disease onset and promoting disease progression in BPH, while ER alpha has inhibitory effect. The activation mechanism of the BPH model in rats and the androgen /AR pathway in human BPH is different from that of the human BPH. In the present study, it is not yet believed that the MI RNAs has a correlation with the abnormal expression of the neutrosteroid receptor in the BPH tissue, nor is it that MI RNAs has a significant role in the pathogenesis of BPH. The difference expression requires a higher sensitivity and specificity, and it is necessary to expand the sample size of the normal prostate to exclude the effect of the sample bias to exclude the effect of the bias of the sample.3. through the present experimental data, it is found that the expression of ETV1 in the BPH tissue is significantly up-regulated at the transcriptional level compared with the normal prostate tissue. There is no statistical significance in the expression of the transcription factors at the transcriptional level. However, the results of this experiment are not yet given, and further studies are needed to clarify the changes in the level of these proteins and the levels of protein phosphorylation. It is preliminarily presumed that the AR-ETV1 pathway may be related to the pathogenesis of BPH and is the possible direction of our in-depth study. .4. confirmed the correlation between MetS and BPH. The total volume of the prostate was larger in patients with BPH with MetS, the level of serum PSA was higher, and the tissue inflammation was more significant. However, it is still not considered that MetS has a significant effect on the related symptoms of BPH. The expression of AR, ER, and PGR in prostate tissue is upregulated, which promotes BPH and disease progression.

【学位授予单位】：第三军医大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R697.3

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