【摘要】： 第一部分蘇州市兒童腸易激綜合征(IBS)的流行病學(xué)危險(xiǎn)因素研究 目的探討蘇州市區(qū)小學(xué)生腸易激綜合征(IBS)的患病率及危險(xiǎn)因素。方法采用流行病學(xué)橫斷面研究方法對(duì)蘇州市滄浪、平江及金閶轄區(qū)內(nèi)公辦小學(xué)1～6年級(jí)小學(xué)生進(jìn)行問(wèn)卷調(diào)查。采用多階段、隨機(jī)整群抽樣的方法獲取調(diào)查樣本。在調(diào)查對(duì)象中,按羅馬Ⅱ診斷標(biāo)準(zhǔn),排除其他器質(zhì)性疾病后確診IBS病例,其他非IBS病例為對(duì)照。擬合非條件logistic多元回歸模型,分析影響IBS發(fā)病可能的危險(xiǎn)因素,為兒童IBS的預(yù)防提供科學(xué)的依據(jù)。結(jié)果共發(fā)放問(wèn)卷8000份,回收有效問(wèn)卷7472份,回收率93.4%;蘇州市區(qū)1～6年級(jí)小學(xué)生有癥狀符合羅馬Ⅱ診斷標(biāo)準(zhǔn)的IBS患病率為10.81%。男、女IBS患病率差異無(wú)統(tǒng)計(jì)學(xué)意義(10.3%vs11.3%)。多因素logistic回歸模型分析提示:有食物過(guò)敏史(OR=1.53,95%CI:1.13～2.07)、兒童期有腸炎史(OR=1.29,95%CI:1.00～1.63)、喜食油炸食物(OR=1.62,95%CI:1.34～1.96)、心情焦慮(OR=1.49,95%CI:1.16～1.93)、兒童幼年時(shí)有意外打擊史(OR=1.47,95%CI:1.02～2.20)及父母有便秘史(OR=1.81,95%CI:1.46～2.24)為男女兒童IBS發(fā)病最可能的危險(xiǎn)因素。兒童年齡越大,IBS發(fā)病危險(xiǎn)有逐漸下降的趨勢(shì)(OR=0.94,95%CI:0.89～0.99)結(jié)論IBS是蘇州市區(qū)小學(xué)生的常見(jiàn)病。影響蘇州市區(qū)小學(xué)生IBS可能的危險(xiǎn)因素是:年齡小、有食物過(guò)敏史、兒童期有腸炎史、喜食油炸食物、心情焦慮、兒童幼年時(shí)有意外打擊史及父母有便秘史。因此,提示臨床醫(yī)師應(yīng)重視此病,積極防治腸炎,調(diào)整兒童飲食,避免多食油炸食品及過(guò)敏的食物,以更好地防治本病。 第二部分幼鼠腸易激綜合征模型的建立及鑒定 目的建立適合兒童腸易激綜合征(IBS)研究的幼鼠IBS動(dòng)物模型并鑒定。方法應(yīng)用20只SD新生大鼠,隨機(jī)分為IBS模型組和正常對(duì)照組,IBS模型組進(jìn)行母子分離、改良機(jī)械束縛、結(jié)腸芥子油刺激等綜合因素干預(yù)幼鼠建立IBS的動(dòng)物模型;正常組不作干預(yù)。用自制的擴(kuò)張器對(duì)幼鼠進(jìn)行直腸擴(kuò)張,評(píng)估不同壓力下腹部收縮反射(AWR)閾值,測(cè)定幼鼠腹壁肌電活動(dòng);利用血管球囊擴(kuò)張導(dǎo)管以5ml、10ml分別擴(kuò)張IBS模型組、對(duì)照組幼鼠直腸,分別采集兩組鼠腦部的fMRI數(shù)據(jù),采用腦功能成像分析軟件進(jìn)行數(shù)據(jù)處理,應(yīng)用血氧水平依賴性功能性MRI(BOLD-fMRI)技術(shù),研究直腸球囊擴(kuò)張刺激下腸易激綜合征模型幼鼠腦內(nèi)臟痛覺(jué)中樞興奮情況及其興奮區(qū)分布情況;取乙狀結(jié)腸降段結(jié)腸用分層刮片技術(shù)刮取由外向內(nèi)取1-4層細(xì)胞,丫啶橙熒光染色觀察幼老細(xì)胞的比值并對(duì)腸粘膜上皮細(xì)胞進(jìn)行美蘭還原反應(yīng)時(shí)間測(cè)定;并行結(jié)腸組織病理檢測(cè);干預(yù)后稱兩組幼鼠體重,觀察糞便性狀,查糞常規(guī),并收集糞便稱濕重和干重,應(yīng)用Bristol分型觀察各組糞便性狀進(jìn)行評(píng)分。結(jié)果直腸擴(kuò)張時(shí),在不同壓力下模型組腹壁收縮反射(AWR)評(píng)分較對(duì)照組閾值顯著降低(P值均0.01);不同擴(kuò)張壓力下,腹壁肌電活動(dòng)隨壓力增加明顯增加(P0.01);直腸球囊刺激后可引起IBS模型組幼鼠腦內(nèi)與內(nèi)臟相關(guān)的腦區(qū)(腦島皮質(zhì)、額前皮質(zhì)、丘腦及扣帶前皮質(zhì))神經(jīng)元活動(dòng)增強(qiáng),表現(xiàn)為高興奮區(qū),而對(duì)照組幼鼠無(wú)相應(yīng)興奮區(qū)表現(xiàn);結(jié)腸黏膜分層刮片后丫啶橙染色見(jiàn)IBS模型組粘膜上皮最外層細(xì)胞幼老細(xì)胞比值較正常對(duì)照組明顯減少,美藍(lán)還原時(shí)間延長(zhǎng)(P0.05);結(jié)腸病理檢查兩組均未見(jiàn)明顯異常;IBS模型組腹瀉癥狀明顯較正常對(duì)照組增加(P0.05),模型組糞便多為軟的團(tuán)塊或泥漿樣,而對(duì)照組為柔軟的香腸狀或團(tuán)塊狀,兩組糞便濕重存在顯著差異(P0.05),而體重、糞便常規(guī)檢查兩組間無(wú)明顯差異。兩組Bristol分型評(píng)分比較有顯著性差異(P0.05)。結(jié)論用母子分離、改良機(jī)械束縛、結(jié)腸芥子油刺激等綜合因素干預(yù)幼鼠可成功建立適合兒童的幼鼠IBS動(dòng)物模型,其機(jī)制可能為內(nèi)臟敏感性增高。功能MRI是一種較為直觀且準(zhǔn)確的觀察腦功能活動(dòng)的腦顯像技術(shù),本研究顯示IBS幼鼠中樞內(nèi)腦島皮質(zhì)、額前皮質(zhì)及丘腦可能是參與內(nèi)臟感覺(jué)的主要部位。幼鼠IBS模型結(jié)腸粘膜上皮細(xì)胞功能處于抑制狀態(tài)。 第三部分ICC細(xì)胞和腦腸肽在幼鼠IBS模型中作用機(jī)制的實(shí)驗(yàn)研究 目的旨在通過(guò)檢測(cè)幼鼠結(jié)腸、脊髓和丘腦組織中腦腸肽NPY和cGRP的mRNA表達(dá)和結(jié)腸ICC細(xì)胞c-kit陽(yáng)性細(xì)胞的表達(dá),探討腦腸肽、腦腸軸及ICC細(xì)胞在幼鼠IBS模型發(fā)病機(jī)制中的作用。方法根據(jù)簡(jiǎn)單數(shù)字法隨機(jī)將乳鼠分成IBS模型組和正常對(duì)照組二組,各10只。IBS模型組用母子分離、改良機(jī)械束縛、結(jié)腸芥子油刺激等綜合因素干預(yù)的方法造模,對(duì)照組則不做任何干預(yù)。采用半定量逆轉(zhuǎn)錄多聚酶鏈?zhǔn)椒磻?yīng)法檢測(cè)二組幼鼠結(jié)腸、脊髓、丘腦組織中NPY和cGRP mRNA的表達(dá)。另取結(jié)腸組織通過(guò)免疫組化檢測(cè)肌間ICC細(xì)胞c-kit陽(yáng)性表達(dá)細(xì)胞數(shù)。結(jié)果IBS模型組中NPY在結(jié)腸、脊髓、丘腦組織中mRNA表達(dá)明顯較正常對(duì)照組降低,t值分別為9.66,8.55,12.65,(均P0.01);而cGRP則在腦腸軸各個(gè)層面(結(jié)腸、脊髓、丘腦)的表達(dá)中,IBS組明顯較正常對(duì)照組增加,t值分別為14.41,12.85,12.41,(均P0.01)。二組幼鼠結(jié)腸ICC細(xì)胞c-kit陽(yáng)性表達(dá)細(xì)胞數(shù)有差異(t=5.294,P0.01)。結(jié)論ICC細(xì)胞、腦腸肽可能在幼鼠IBS模型的發(fā)生、發(fā)展中起重要作用,本研究為深入研究?jī)和疘BS發(fā)病機(jī)制提供了實(shí)驗(yàn)依據(jù)。 第四部分馬來(lái)酸曲美布汀干預(yù)幼鼠IBS模型的療效觀察 目的探討馬來(lái)酸曲美布汀干預(yù)幼鼠腸易激綜合征(IBS)模型后的治療療效。方法用母子分離、改良機(jī)械束縛、結(jié)腸芥子油刺激等綜合的方法建立幼鼠IBS模型共20只,隨機(jī)選10只在干預(yù)同時(shí)用馬來(lái)酸曲美布汀3mg/kg.d灌服治療為治療組,另10只為模型組,取同期正常生長(zhǎng)幼鼠10只為正常對(duì)照組。分別對(duì)三組幼鼠行肛門直腸球囊擴(kuò)張刺激下測(cè)腹壁肌電活動(dòng),AWR評(píng)分;處死后取結(jié)腸對(duì)其腸粘膜上皮細(xì)胞進(jìn)行分層刮片、熒光染色和還原能力測(cè)定,RT-PCR檢測(cè)結(jié)腸、脊髓、丘腦中NPY和cGRP的mRNA表達(dá)。結(jié)果IBS模型組和正常對(duì)照組間不同壓力下AWR評(píng)分均有顯著性差異(P0.05),而治療組和正常對(duì)照組間差異無(wú)顯著性(P0.05);在8mmHg壓力時(shí),IBS模型組腹壁收縮次數(shù)增加達(dá)(6.65±1.04)次/3 min ,肌電圖振幅增高,與另二組比較有顯著性(P0.01),而治療組、正常對(duì)照組間腹壁收縮活動(dòng)和肌電圖振幅差異無(wú)顯著性(p0.05);在12mmHg、15mmHg擴(kuò)張壓力下,IBS模型組腹壁收縮活動(dòng)增加,肌電圖振幅增高(P0.05),但正常對(duì)照組與治療組間無(wú)顯著差異(p0.05);在28mmHg壓力擴(kuò)張下,三組幼鼠腹壁收縮次數(shù)及肌電圖振幅增高差異均無(wú)顯著性(P0.05);三組結(jié)腸粘膜由外向內(nèi)1-4層取材標(biāo)本中,IBS模型組的腸粘膜上皮細(xì)胞美藍(lán)還原反應(yīng)時(shí)間最長(zhǎng)(11.8±1.45)min ;NPY在IBS模型組結(jié)腸、脊髓、丘腦三個(gè)部位的表達(dá)均較另二組低,F值分別為11.29,34.15,27.13(均P0.05),而治療組和對(duì)照組間差異無(wú)顯著性;cGRP在模型組三個(gè)部位的表達(dá)均較正常對(duì)照組和治療組增強(qiáng),F值分別為98.58,39.12,82.67(均P0.01),而治療組和對(duì)照組間表達(dá)無(wú)顯著性差異(P0.05)。結(jié)論馬來(lái)酸曲美布汀治療幼鼠IBS模型有效,可改善其腸道運(yùn)動(dòng)功能及內(nèi)臟高敏感性。
[Abstract]:Part one: epidemiological risk factors for irritable bowel syndrome (IBS) in children in Suzhou.
Objective To investigate the prevalence and risk factors of irritable bowel syndrome (IBS) among pupils in Suzhou. Methods A cross-sectional epidemiological study was conducted among the pupils of grade 1 to 6 in Canglang, Pingjiang and Jinlong primary schools. According to Rome II diagnostic criteria, IBS cases were diagnosed after excluding other organic diseases, and other non-IBS cases were compared. Unconditional logistic multiple regression model was fitted to analyze the possible risk factors affecting the incidence of IBS and provide scientific basis for the prevention of IBS in children. There was no significant difference in the prevalence of IBS between males and females (10.3% vs 11.3%). Multivariate logistic regression analysis showed that there was a history of food allergy (OR = 1.53, 95% CI: 1.13-2.07), a history of enteritis in childhood (OR = 1.29, 95% CI: 1.00-1.63), and a preference for food. Fried food (OR = 1.62,95% CI: 1.34-1.96), mood anxiety (OR = 1.49,95% CI: 1.16-1.93), history of unintentional attack in childhood (OR = 1.47,95% CI: 1.02-2.20) and history of constipation in parents (OR = 1.81,95% CI: 1.46-2.24) were the most likely risk factors for IBS in boys and girls. Conclusion IBS is a common disease among pupils in Suzhou. The possible risk factors affecting IBS among pupils in Suzhou are: younger age, history of food allergy, history of enteritis in childhood, preference for fried food, anxiety, history of accidental attack in childhood and history of constipation in parents. We should actively prevent and treat enteritis, adjust children's diet, avoid eating more fried food and allergic food, so as to prevent and cure the disease better.
The second part is the establishment and identification of irritable bowel syndrome model in young rats.
Methods Twenty SD neonatal rats were randomly divided into IBS model group and normal control group. The infant rats in IBS model group were intervened by maternal-fetal separation, improved mechanical restraint and colon mustard oil stimulation to establish an IBS animal model. Intervention: Rectal dilatation was performed with a self-made dilator to evaluate the abdominal contractile reflex (AWR) threshold under different pressures, and the abdominal wall electromyographic activity was measured. The IBS model group was dilated with a balloon dilator catheter of 5ml and 10ml respectively, and the control group was dilated with rectum. The fMRI data of the brains of the two groups were collected and analyzed by functional brain imaging. Data were processed by software and blood oxygen level-dependent functional MRI (BOLD-fMRI) technique was used to study the excitability of visceral pain center and the distribution of excitatory areas in the brains of irritable bowel syndrome (IBS) rats induced by rectal balloon dilatation. The ratio of young and old cells was observed by fluorescence staining and the methylene blue reduction reaction time of intestinal mucosal epithelial cells was determined. The colon histopathological examination was carried out. After intervention, the body weight of the two groups of young rats was weighed, the stool characteristics were observed, the stool routine was checked, and the wet and dry weights were collected. The stool characteristics of each group were scored by Bristol typing. The abdominal wall systolic reflex (AWR) score in the model group was significantly lower than that in the control group at different pressure (P 0.01); the abdominal wall electromyographic activity was significantly increased with the increase of pressure (P 0.01); rectal balloon stimulation could induce the viscera-related brain areas (insular cortex, prefrontal cortex, thalamus and anterior cortex) in the brain of the IBS model group. The activity of neurons in the anterior cingulate cortex was enhanced, showing the excitatory zone, but there was no corresponding excitatory zone in the control group. The symptoms of diarrhea in IBS model group were significantly higher than those in normal control group (P There was significant difference (P 0.05). Conclusion Infant mice with IBS can be successfully established by the intervention of maternal-fetal separation, modified mechanical restraint and colon mustard oil stimulation. The mechanism may be increased visceral sensitivity. Functional MRI is a more intuitive and accurate brain imaging technique for observing brain function. It is suggested that the insular cortex, prefrontal cortex and thalamus may be the main sites involved in visceral sensation in young rats with IBS.
The third part is the experimental study on the mechanism of ICC cell and brain gut peptide in the IBS model of young rats.
Objective To investigate the role of brain-gut peptide, brain-gut axis and ICC cells in the pathogenesis of infant rat IBS model by detecting the expression of brain-gut peptide NPY and cGRP mRNA in colon, spinal cord and thalamus and the expression of c-kit positive cells in colon ICC cells. Rats in the IBS model group were established by the method of maternal-fetal separation, modified mechanical restraint and colon mustard oil stimulation, while those in the control group were given no intervention. The expressions of NPY and cGRP mRNA in colon, spinal cord and thalamus were detected by Semi-quantitative reverse transcriptase polymerase chain reaction. Results The expression of NPY in colon, spinal cord and thalamus of IBS model group was significantly lower than that of normal control group, with T values of 9.66, 8.55 and 12.65 respectively (all P 0.01), while the expression of cGRP in all levels of brain-gut axis (colon, spinal cord, thalamus) was significantly higher in IBS model group than that of normal control group. There were significant differences in the number of c-kit positive cells between the two groups (t = 5.294, P 0.01). Conclusion ICC cells and brain-gut peptides may play an important role in the development of IBS model in young rats. This study provides experimental basis for further study of the pathogenesis of IBS in children.
The fourth part of trimebutine maleate intervened in the IBS model of young rats.
Objective To investigate the therapeutic effect of trimebutine maleate on irritable bowel syndrome (IBS) in young rats. Methods Twenty IBS models were established by maternal-fetal separation, modified mechanical restraint and colon mustard oil stimulation. Ten rats were randomly selected as the treatment group and 10 rats were treated with trimebutine maleate at the same time. The abdominal myoelectric activity and AWR score were measured under anorectal balloon dilatation stimulation, and the intestinal mucosal epithelial cells were scraped, stained and reduced by fluorescence. NPY and cGRP in colon, spinal cord, thalamus were detected by RT-PCR. Results There was significant difference in AWR score between IBS model group and normal control group (P 0.05), but there was no significant difference between treatment group and normal control group (P 0.05). At 8 mmHg pressure, the number of abdominal wall contractions in IBS model group increased by (6.65 (1.04) times/3 minutes, and the amplitude of electromyogram increased significantly (P 0.0). 1) There was no significant difference in abdominal wall contraction and EMG amplitude between the treatment group and the control group (p0.05); the abdominal wall contraction and EMG amplitude increased in the IBS model group under 12 mmHg and 15 mmHg dilatation pressure (P 0.05), but there was no significant difference between the normal control group and the treatment group (p 0.05); the abdominal wall contraction of the three groups of young rats under 28 mmHg dilatation pressure. There was no significant difference in frequency and amplitude of EMG (P 0.05). Among the three groups, methylene blue reduction reaction time of intestinal epithelial cells in IBS model group was the longest (11.8 The expression of cGRP in the three parts of the model group was higher than that of the normal control group and the treatment group, F values were 98.58, 39.12 and 82.67 respectively (all P 0.01), but there was no significant difference between the treatment group and the control group (P 0.05). Conclusion Trimebutine maleate is effective and can be modified in the treatment of IBS model in young rats. Good bowel movement function and visceral Gao Min sensibility.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R725.7;R-332

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