本文選題：前列化濁方 + 前列腺炎　； 參考：《中國中醫(yī)科學(xué)院》2014年碩士論文

【摘要】：慢性前列腺炎(chronic prostatitis,CP)是中青年男性的常見病和多發(fā)病,約占泌尿外科門診患者的33%。該病纏綿難愈、反復(fù)發(fā)作,給患者造成很大的生理痛苦及心理負(fù)擔(dān)。美國國立衛(wèi)生研究院已將前列腺炎、心肌梗死、不穩(wěn)定性心絞痛和活動(dòng)性Crohn病等一起列為影響居民生活質(zhì)量最為嚴(yán)重的慢性疾病。然而,目前其發(fā)病機(jī)制尚未完全闡明,大量研究表明多種細(xì)胞因子構(gòu)成的網(wǎng)絡(luò)和級(jí)聯(lián)反應(yīng)可能是導(dǎo)致CP的主要原因。細(xì)胞因子信號(hào)轉(zhuǎn)導(dǎo)抑制蛋白(suppressor of cytokine signal,SOCS)能夠調(diào)節(jié)細(xì)胞對(duì)細(xì)胞因子的反應(yīng)。但是,SOCS在本病發(fā)生發(fā)展過程中發(fā)揮的作用至今尚未得到論證。 導(dǎo)師既往臨床研究證實(shí),前列化濁方(黃柏、茵陳、赤芍、紅花、三七、柴胡、香附)在改善前列腺炎患者癥狀,提高患者生活質(zhì)量方面具有滿意的療效和顯著的優(yōu)勢。很多學(xué)者研究認(rèn)為前列腺炎是一種自身免疫性疾病�；诖�,本研究以實(shí)驗(yàn)性自身免疫性前列腺炎(experimental autoimmune prostatitis,EAP)大鼠模型為載體,通過前列化濁方的干預(yù),采用RT-PCR和Western-blot方法檢測大鼠前列腺組織相關(guān)細(xì)胞因子及SOCS的表達(dá),揭示細(xì)胞因子與SOCS在CP發(fā)生發(fā)展中的共同作用,闡明前列化濁方治療CP的作用機(jī)理,為臨床應(yīng)用前列化濁方提供實(shí)驗(yàn)依據(jù)。 前列腺炎和良性前列腺增生癥(benign prostatic hyperplasia,BPH)常同時(shí)存在,相互影響。盡管有臨床研究報(bào)道慢性炎癥在BPH發(fā)病中發(fā)揮重要作用,與未患有前列腺炎男性相比,患有CP的患者更易發(fā)展為BPH。但是,目前CP和BPH的相關(guān)性研究大多以BPH患者為研究對(duì)象,研究者將BPH患者前列腺切除后獲取的標(biāo)本行病理學(xué)檢查,以分析炎癥和BPH的相互關(guān)系。有學(xué)者認(rèn)為這類研究屬回顧性研究,未能明確CP和BPH的發(fā)病順序,也未能排除年齡在BPH發(fā)病中的影響。所以炎癥和BPH之間的關(guān)系尚不明確,炎癥在BPH進(jìn)展中的作用機(jī)制尚缺乏強(qiáng)有力的證據(jù)。 Biochim Biophys Acta雜志2013年1月刊出Kim HJ等采用脂多糖注射大鼠前列腺方法建立前列腺炎大鼠模型,研究CP導(dǎo)致BPH的機(jī)制。鑒于此,本研究在國內(nèi)率先以EAP大鼠模型為載體,采用前瞻性研究方法初步探索炎癥在BPH進(jìn)展中的作用機(jī)制,既可明確炎癥與BPH的發(fā)病順序,也能夠排除年齡在BPH進(jìn)展中的影響,有利于揭示炎癥在BPH進(jìn)展中的作用機(jī)制及闡明前列化濁方干預(yù)BPH進(jìn)展的機(jī)理。 國內(nèi)外均有學(xué)者認(rèn)為有效防治前列腺炎,可以降低BPH的發(fā)生和減緩BPH的進(jìn)展,慢性非細(xì)菌性前列腺炎為防治BPH提供了新靶點(diǎn),為了延緩或阻斷BPH發(fā)生發(fā)展,未來的研究應(yīng)主要集中于細(xì)胞因子,它可能成為潛在的治療靶點(diǎn)。故本研究以細(xì)胞因子、SOCS、Bcl-2和病理形態(tài)學(xué)等多角度多層次開展深入研究,以期揭示炎癥在BPH進(jìn)展中的作用機(jī)制,干預(yù)BPH的發(fā)生發(fā)展。 本研究為導(dǎo)師承擔(dān)的國家自然基金面上項(xiàng)目(81072814),研究分為兩部分：文獻(xiàn)綜述和動(dòng)物實(shí)驗(yàn)研究。在導(dǎo)師指導(dǎo)下,我參與了課題前期準(zhǔn)備、建立動(dòng)物模型、灌胃給藥、實(shí)驗(yàn)取材、統(tǒng)計(jì)分析等工作。 1.文獻(xiàn)綜述：本部分分別從NIH-Ⅲ型前列腺炎與相關(guān)細(xì)胞因子的研究進(jìn)展,炎癥在BPH進(jìn)展中作用機(jī)制的研究進(jìn)展兩個(gè)方面進(jìn)行綜述。 2.動(dòng)物實(shí)驗(yàn)研究：炎癥對(duì)前列腺增生的影響及前列化濁方的干預(yù)效應(yīng)研究 目的：觀察前列化濁方對(duì)EAP大鼠的干預(yù)效應(yīng)。 方法：取240-300gSD雄性大鼠66只,隨機(jī)分為正常組12只,實(shí)驗(yàn)組54只。采用注射自身抗原的方法建立實(shí)驗(yàn)性自身免疫性前列腺炎(experimental autoimmune prostatitis,EAP)大鼠模型,實(shí)驗(yàn)組分別于0、7、14、21天腹腔注射百白破疫苗0.5ml,并同時(shí)皮內(nèi)多點(diǎn)注射大鼠前列腺蛋白提純液(濃度15g/L)和福式完全佐劑的混懸液(比例1：1)1.0ml。正常組分別行腹腔及皮內(nèi)多點(diǎn)注射0.9%生理鹽水注射液0.5、1.0ml。在第35天,實(shí)驗(yàn)組處死6只大鼠,將前列腺組織送西苑醫(yī)院病理室檢驗(yàn),病理結(jié)果證實(shí)成功建立了前列腺炎大鼠模型。剩余48只大鼠隨機(jī)分為模型組、前列化濁方低劑量組(以下簡稱低劑量組)、前列化濁方中劑量組(以下簡稱中劑量組)、前列化濁方高劑量組(以下簡稱高劑量組),每組12只大鼠。除模型組和正常組給予蒸餾水外,低劑量組、中劑量組、高劑量組分別給予低劑量前列化濁方(按5.25g/kg體重給藥)、中劑量前列化濁方(按10.5g/kg體重給藥)和高劑量前列化濁方(按21.0g/kg體重給藥),每天灌胃給藥一次,連續(xù)干預(yù)30天后,腹腔注射20%烏拉坦(用量按體重計(jì)算,0.4ml/100g)麻醉大鼠。局部皮膚消毒,在無菌條件下,下腹部正中切口,根據(jù)膀胱及精囊的位置找到前列腺,取出前列腺組織,剪除多余脂肪組織,用冷生理鹽水洗凈,摘取左側(cè)葉前列腺分別置固定液中,用于常規(guī)HE染色,光鏡下觀察各組大鼠前列腺腺腔及間質(zhì)的炎性細(xì)胞浸潤和增生情況。迅速取右側(cè)前列腺組織置液氮中,用于RT-PCR和Western-blot檢測。RT-PCR法測定大鼠前列腺組織Interleukin-1(IL-1)、Interleukin-6(IL-6)、Interleukin-10(IL-10) Interleukin-17(IL-17)、Tumor necrosis factor-a(TNF-a)和B cell leukemia-2(Bcl-2)的mRNA表達(dá)水平,Western-blot法檢測大鼠前列腺組織SOCS-1和SOCS-3表達(dá)水平。 結(jié)果： (1)組織病理學(xué)觀察顯示：模型組腺體排列紊亂,緊密,大量腺體呈乳頭狀增生,腺體上皮由立方狀變?yōu)楦咧鶢?腺腔內(nèi)可見中性粒細(xì)胞；間質(zhì)增生,血管擴(kuò)張充血,組織水腫,大量中性粒細(xì)胞及少量淋巴細(xì)胞浸潤。低劑量組、中劑量組和高劑量組炎癥和腺體增生均呈不同程度減輕。組織病理學(xué)結(jié)果證實(shí)：成功建立了EAP大鼠模型,前列化濁方具有治療前列腺炎和干預(yù)BPH進(jìn)展的作用。 (2)炎癥因子檢測顯示：與正常組比較,模型組大鼠前列腺組織IL-10表達(dá)下降(P0.01),IL-1、IL-6、IL-17和]NF-α表達(dá)均增高(P0.01)。表明EAP大鼠前列腺組織存在強(qiáng)烈的炎癥反應(yīng)。 與模型組比較,治療組(低劑量組、中劑量組和高劑量組)大鼠前列腺組織IL-6、IL-17和TNF-α的表達(dá)均有下降(P0.01)；其中,中劑量組下降尤為顯著(P0.01)。 與模型組比較,IL-1在中劑量組大鼠前列腺組織表達(dá)下降明顯(P0.01),但高劑量組和低劑量組前列腺組織中IL-1水平與模型組比較無顯著性差異(P0.05)。 與模型組比較,治療組大鼠前列腺組織IL-10表達(dá)均上升(P0.01),中劑量組和低劑量組上升明顯。提示前列化濁方對(duì)炎癥因子的影響可能與劑量有關(guān)。 (3)凋亡抑制因子Bcl-2檢測顯示：與正常組比較,模型組大鼠前列腺組織Bcl-2的表達(dá)明顯升高(P0.01)；與模型組比較,中劑量組大鼠前列腺組織Bcl-2的表達(dá)明顯降低(P0.01)。 (4)SOCS-1和SOCS-3檢測顯示：與正常組比較,模型組大鼠前列腺組織SOCS-1和SOCS-3表達(dá)顯著升高(P0.01)；與模型組比較,治療組大鼠前列腺組織SOCS-1和SOCS-3表達(dá)均下降(P0.01),中劑量組下降最顯著。 結(jié)論： 模型組大鼠病理形態(tài)學(xué)檢測結(jié)果及前列腺組織炎癥因子的表達(dá)水平均證實(shí)成功建立了EAP大鼠模型；治療組大鼠前列腺組炎癥和增生緩解,從形態(tài)學(xué)證實(shí)了前列化濁方治療CP和干預(yù)BPH進(jìn)展的療效。 模型組大鼠前列腺組織前炎癥因子表達(dá)顯著高于正常組,治療組大鼠前列腺組織前炎癥因子表達(dá)顯著低于模型組。提示前列化濁方可能通過抑制前炎癥細(xì)胞因子表達(dá)發(fā)揮治療作用。 模型組大鼠前列腺組織IL-10表達(dá)顯著低于正常組,治療組大鼠前列腺組織IL-10表達(dá)顯著高于模型組。提示前列化濁方治療CP的作用可能通過升高抗炎癥因子表達(dá)和調(diào)節(jié)免疫功能實(shí)現(xiàn)的。 模型組大鼠前列腺組織前炎癥因子、SOCS-1和SOCS-3表達(dá)顯著高于正常組,證實(shí)炎癥因子與SOCS可能共同影響著CP的發(fā)生發(fā)展,JAK/STAT信號(hào)通路在CP的發(fā)生發(fā)展中可能扮演了重要的角色,有待深入研究。 模型組大鼠前列腺組織Bcl-2表達(dá)顯著高于正常組,中劑量組大鼠前列腺組織Bcl-2表達(dá)顯著低于模型組。病理形態(tài)學(xué)檢測顯示模型組大鼠前列腺組織大量腺體呈乳頭狀增生,腺體上皮由立方狀變?yōu)楦咧鶢�；治療組大鼠前列腺極少量腺體乳頭狀增生,間質(zhì)增生不明顯。提示炎癥在BPH進(jìn)展中可能扮演重要角色,前列化濁方可能通過減輕前列腺組織炎癥反應(yīng)和下調(diào)Bcl-2表達(dá)干預(yù)BPH進(jìn)展。
[Abstract]:Chronic prostatitis (chronic prostatitis, CP) is a common and frequently occurring disease of young and middle-aged men. It accounts for the 33%. of the outpatients in the Department of Urology, which is difficult to recover and relapse, causing great physical pain and psychological burden to the patients. The National Institutes of health of the United States has put forward the prostatitis, myocardial infarction, unstable angina and active C. Rohn disease, etc., is listed as the most serious chronic disease affecting the quality of life of the residents. However, its pathogenesis has not yet been fully elucidated. A large number of studies have shown that the network and cascade reaction of multiple cytokines may be the main causes of CP. The cytokine signal transduction inhibitor (suppressor of cytokine signal, SOCS) can be the main cause of the disease. Regulation of cell response to cytokines. However, the role of SOCS in the pathogenesis of this disease has not yet been demonstrated.
Previous clinical studies have confirmed that prostoriate turbid recipe (cypress, Artemisia, red peony, red flower, 37, bupleurum, and fragrant) has a satisfactory effect and significant advantage in improving the symptoms of prostatitis and improving the quality of life of the patients. Many scholars have studied that prostatic inflammation is a autoimmune disease. Based on this, this study is experimental The rat model of experimental autoimmune prostatitis (EAP) was used as a carrier. Through the intervention of Prostant turbid recipe, the expression of cytokines and SOCS in the prostate tissue of rats was detected by RT-PCR and Western-blot methods. The common effect of cytokines and SOCS in the development of CP was revealed, and the prostad turbid recipe was clarified. The therapeutic mechanism of CP is to provide experimental evidence for clinical application of prostaglandin turbid prescription.
Prostatitis and benign prostatic hyperplasia (benign prostatic hyperplasia, BPH) often exist and interact with each other. Although there are clinical reports that chronic inflammation plays an important role in the pathogenesis of BPH, patients with CP are more likely to develop into BPH. than those with no prostatitis, but the current correlation of CP and BPH is mostly BPH. The patients were studied. The researchers examined the pathological examination of specimens obtained after the prostatectomy in BPH patients to analyze the relationship between inflammation and BPH. Some scholars believe that this kind of study is a retrospective study that does not define the sequence of CP and BPH, nor does it exclude the influence of age in the pathogenesis of BPH. So there is no relationship between inflammation and BPH. It is clear that there is no strong evidence for the role of inflammation in the progression of BPH.
Biochim Biophys Acta magazine published Kim HJ and Kim HJ in January 2013 to establish prostatitis model in rats by injecting lipopolysaccharide into the prostatitis rat model and study the mechanism of BPH caused by CP. In view of this, this study took the lead in the domestic EAP rat model as the carrier, and explored the mechanism of inflammation in the progression of BPH by prospective study methods. It is true that inflammation and the sequence of BPH can also exclude the influence of age on the progress of BPH, and to reveal the mechanism of inflammation in the progress of BPH and to clarify the mechanism of the intervention of prostaglandin turbid prescription in the progress of BPH.
Both domestic and foreign scholars believe that effective prevention and control of prostatitis can reduce the occurrence of BPH and slow down the progress of BPH. Chronic non bacterial prostatitis provides a new target for the prevention and control of BPH. In order to delay or block the development of BPH, the future research should focus mainly on the cytokine, and it may be a potential target for treatment. In order to reveal the mechanism of inflammation in the progress of BPH and to interfere with the development of BPH, many angles and multiple levels, such as cytokine, SOCS, Bcl-2 and Pathomorphology, are carried out in depth.
This study is the national natural fund project (81072814) undertaken by the tutor. The study is divided into two parts: literature review and animal experiment. Under the guidance of the tutor, I participated in the preparation of the project, the animal model, the gavage, the experimental materials, the statistical analysis and so on.
1. literature review: this part is a review of the progress of NIH- III prostatitis and related cytokines, and the progress of the research on the mechanism of inflammation in the progress of BPH.
2. animal experimental study: the effect of inflammation on benign prostatic hyperplasia and the intervention effect of prostaglandin turbid recipe
Objective: To observe the intervention effect of Qian lie Hua Zhuo Fang on EAP rats.
Methods: 66 male 240-300gSD rats were randomly divided into 12 rats in the normal group and 54 in the experimental group. The experimental autoimmune prostatitis (experimental autoimmune prostatitis, EAP) rat model was established by injection of self antigen. The experimental group was injected with a pertussis vaccine 0.5ml in the abdominal cavity on 0,7,14,21 days, and the Pinedo point was given at the same time. The rat prostatic protein purification fluid (concentration 15g/L) and the mixed suspension of full adjuvant (1:1) in the normal group were injected into the abdominal cavity and the intraperitoneal injection of 0.9% saline injection of 0.9% physiological saline injection 0.5,1.0ml. for thirty-fifth days. The experimental group died of 6 rats. The prostate tissue was sent to the Xiyuan Hospital pathology laboratory, and the pathological results confirmed the successful establishment of the test. The remaining 48 rats were randomly divided into model group, low dose group (hereinafter referred to as low dose group), low dose group of prostad turbid prescription (hereinafter referred to as low dose group), middle dose group (hereinafter referred to as middle dose group), high dose group (hereinafter referred to as high dose group), each group of high dose group (hereinafter referred to as high dose group), each group was given the low dose group except the model group and the normal group. In the middle dose group, the high dose group was given a low dose of prostapido turbid (5.25g/kg weight), the middle dose of prostapido turbid prescription (according to 10.5g/kg weight) and the high dose of prostachiated turbid prescription (according to 21.0g/kg weight) was administered every day, and after 30 days of continuous intervention, the intraperitoneal injection of 20% urantan (the dosage according to weight, 0.4ml/100g) anesthesia Intoxicated rats, local skin disinfection, under the aseptic condition, the lower abdomen is median incision, according to the location of the bladder and seminal vesicle to find the prostate, remove the prostate tissue, remove the excess fat tissue, clean the prostate with cold physiological saline, and take the left lobe prostate respectively in the fixed solution, which is used for routine HE staining, and the prostate gland cavity of each group is observed under light microscope. RT-PCR and Western-blot were used to detect Interleukin-1 (IL-1) in the prostate tissue of rats, Interleukin-6 (IL-6), Interleukin-10 (IL-10) Interleukin-17 (IL-17). The expression level of mRNA and Western-blot were used to detect the expression level of SOCS-1 and SOCS-3 in rat prostate tissue.
Result錛，

本文編號(hào)：1836117