儿童功能性消化不良流行病学和发病机制的研究

发布时间：2018-03-04 03:20

本文选题：功能性消化不良　切入点：流行病学　出处：《苏州大学》2012年博士论文　论文类型：学位论文

【摘要】：研究背景随着医学模式由单一的生物模式向生物—心理—社会模式的转变，功能性胃肠病(functional gastrointestinal disorders,FGIDs)越来越受到人们的重视，功能性消化不良（functional dyspepsia，FD）是最常见的功能性胃肠病之一，是一组以反复发作的餐后饱胀、早饱、厌食、嗳气、恶心、呕吐、上腹痛、上腹烧灼感或反酸为主要表现而经各项检查排除器质性、系统性或代谢性疾病的一组常见临床症候群。其解剖定位为胃及十二指肠。罗马委员会根据主要症状的不同将FD分为餐后不适综合征（postprandial distress syndrome, PDS）和上腹痛综合征（epigastric painsyndrome, EPS）两个亚型，越来越多的证据表明FD存在不同的亚型，其临床表现和发病机制不尽相同，建议对不同亚型进行独立评估和治疗。功能性消化不良在全球范围内发病十分普遍，国外文献报道，普通人群中FD患病率约26～34%，而国内尚缺乏较系统的儿童FD流行病学调查。迄今为止，其病因及病理生理机制尚不清楚，近年来研究发现幽门螺杆菌(Helicobacter pylori，Hp)、胃肠激素、炎症细胞等在FD的发病中起重要作用。目的本课题研究目的：了解苏州市儿童FD的患病率及相关因素，调查FD儿童的Hp感染率；研究胃肠激素Ghrelin、Leptin及PYY在儿童FD发病机制中的作用；探讨嗜酸性粒细胞及趋化因子Eotaxin和RANTES与儿童FD临床症状的相关性。为临床儿童功能性消化不良病理生理机制的进一步阐明和临床开发治疗FD的新型靶药提供理论依据。方法 1.流行病学调查： 2010年10～12月对苏州市沧浪区22所小学12923名小学生进行横断面调查，其中男生7265名，女生5658名，年龄6～13岁。FD诊断符合罗马Ⅲ标准，并将FD分为PDS和EPS两个亚型，采用问卷调查和体格检查的方法调查儿童FD的患病率及相关因素，比较不同年龄、不同性别之间FD的患病率是否存在差异,诊断为FD的患儿，进行13C呼气试验检查Hp感染。 2. ELISA检测血浆中Ghrelin、Leptin及PYY水平： 124例FD患儿（男71例，女53例，年龄7～12岁）和27例健康儿童（男15例，女12例，年龄7～12岁）纳入本项研究。FD组根据主要症状和是否伴Hp感染分为PDS组61例，EPS组63例； Hp+和Hp-各62例。入选儿童均在上午8～10时采集清晨空腹静脉血2ml,注入含有EDTA和抑肽酶的试管中，充分混匀离心后收集血浆，-70℃低温冰箱冻存。标本收齐后用ELISA方法检测血浆中Ghrelin、Leptin及PYY水平，并比较各组结果是否存在差异。 3.RT-PCR法检测胃黏膜Ghrelin、Leptin及PYY的mRNA表达： 43例FD患儿（Hp阳性20例，Hp阴性23例）纳入本研究，在上午8～10时行常规胃镜检查，于胃窦部取粘膜两块，一块做快速尿素酶试验检测Hp感染，另一块放入EP管后，于-70℃低温冰箱冻存，待标本集齐后,RT-PCR法检测Ghrelin、Leptin及PYY的mRNA表达,并比较Hp+组和Hp-组的结果是否有差异。 4.HE染色和免疫组化： 71例FD患儿，Hp+23例，Hp-48例（包括EPS25例、PDS23例），21例健康儿童为对照组。取胃窦粘膜固定、切片、HE染色后选取5个清晰的不同高倍视野，400倍光镜下统计嗜酸性粒细胞的数量并求和。采用免疫组化方法检测胃粘膜中趋化因子Eotaxin和RANTES的表达，每张切片随机选取3个有代表性的视野，400倍光镜下测量并计算每张切片免疫组化阳性染色的平均灰度值，并比较各组之间的结果。结果 1.流行病学调查结果：(1)调查12923例儿童，诊断FD1034例，其中EPS646例，PDS388例，苏州市儿童FD总的患病率为8%。其中EPS为5%，PDS为3%。(2) FD儿童1034例经13C呼气试验检查，诊断Hp阳性282例，Hp的感染率为27.3%。(3) PDS患病率的相关因素:年龄越小，父母文化程度越低，龋齿越多，体型偏瘦，患病率越高。(4) EPS患病率的相关因素:女性，龋齿越多，父亲文化程度越高，体型消瘦或超重,患病率越高。 2.血浆中Ghrelin水平：FD组低于对照组，PDS组低于EPS组，Hp+组低于Hp-组，差异均有统计学意义(P＜0.001）；血浆Leptin水平：FD组高于对照组，PDS组高于EPS组，Hp+组高于Hp-组，差异均有统计学意义(P＜0.001)；血浆中PYY水平：FD组高于对照组，差异有统计学意义(P＜0.001)，PDS组与EPS组，Hp+组与Hp-组比较，差异无统计学意义。 3.胃粘膜中， Hp+组ghrelin mRNA水平显著低于Hp-组，Hp+组leptinmRNA水平显著高于Hp-组的表达，差异均有统计学意义(P＜0.01)， PYY在儿童胃粘膜中几乎未见表达。 4.胃粘膜嗜酸性粒细胞计数、Eotaxin和RANTES表达水平在Hp+和EPS/Hp-组明显高于PDS/Hp-组和对照组，差异均有统计学意义(P＜0.001)。趋化因子Eotaxin主要分布在胃黏膜的上皮细胞和腺体上皮细胞的胞浆内，而炎症细胞内未见分布。RANTES在胃黏膜主要分布于胃粘膜上皮细胞、腺体细胞、淋巴细胞的细胞浆和细胞核内，Eotaxin和RANTES表达水平与嗜酸性粒细胞数量呈正相关结论 1.FD是苏州地区儿童的常见病，主要亚型是EPS。儿童FD发病的危险因素有女性、年龄小、父母文化程度、Hp感染、龋齿、BMI降低。 2Ghrelin、Leptin及PYY三种胃肠激素参与了儿童功能性消化不良的发病机制,Ghrelin和Leptin在餐后不适（PDS）亚型中变化更加明显，推测PDS早饱、厌食等症状与Ghrelin和Leptin有更加直接的关系，Ghrelin可能成为治疗PDS的新型靶药。 3.嗜酸性粒细胞及趋化因子eotaxin和RANTES主要参与了儿童功能性消化不良上腹痛（EPS）的发病和疾病进展，为临床研发治疗EPS的新型药物提供了实验依据。
[Abstract]:Research background
With the medical model from a single biological model to bio psycho social model, functional gastrointestinal disease (functional gastrointestinal, disorders, FGIDs) has attracted more and more attention, functional dyspepsia (functional dyspepsia FD) is one of the most common functional gastrointestinal disorders, is a recurrent meal after the fullness, early satiety, belching, nausea, anorexia, vomiting, epigastric pain, epigastric burning sensation or acid reflux is the main and the performance of the examination to exclude organic system, or a group of metabolic diseases common clinical syndrome. The anatomic location for the stomach and duodenum. The Rome committee will be divided into FD postprandial distress syndrome according to the main symptoms of the different (postprandial distress syndrome, PDS) and epigastric pain syndrome (epigastric, painsyndrome, EPS) two subtypes, there is growing evidence that FD has its different subtypes. The clinical manifestations and pathogenesis are different. It is suggested that different subtypes should be evaluated and treated independently.
Functional dyspepsia worldwide disease is very common, foreign literature reports, the prevalence rate of about 26 ~ 34% FD in the general population, and the lack of children's FD epidemiological survey systematically. So far, the etiology and pathophysiology remains unclear, recent studies found that Helicobacter pylori (Helicobacter pylori, Hp), gastrointestinal hormones, inflammatory cells play an important role in the pathogenesis of FD.
objective
This study is to investigate the prevalence rate and related factors of children in Suzhou FD children, investigation of FD infection rate of Hp; Study on gastrointestinal hormones Ghrelin, Leptin and PYY in the pathogenesis of FD in children; to investigate the relationship between eosinophil and chemokine Eotaxin and RANTES in children with FD clinical symptoms. The theoretical basis of new target drugs and further clarify the clinical development for clinical treatment of FD in children with functional dyspepsia pathophysiology.
Method
1. epidemiological survey:
In 2010 10~12 a cross-sectional survey was conducted in 22 primary school in Canglang District of Suzhou City, 12923 students, including 7265 boys and 5658 girls, aged 6~13 years old.FD diagnostic Rome III standard and FD were divided into PDS and EPS two subtypes, the prevalence rate and related factors of questionnaire survey and physical examination method the survey of children FD, different age, different gender differences exist between the FD prevalence rate is, children who were diagnosed FD, 13C breath test for Hp infection.
The levels of Ghrelin, Leptin and PYY in plasma were measured by 2. ELISA.
In 124 FD patients (male 71 cases, female 53 cases, age 7~12 years) and 27 healthy children (male 15 cases, female 12 cases, age 7~12) into the.FD group in this study according to the main symptoms and with Hp infection were divided into PDS group of 61 cases, 63 cases in group EPS; Hp+ and Hp- the 62 cases of children were selected. Fasting venous blood 2ml in the afternoon 8~10 when collecting, injected with EDTA and aprotinin in vitro, and mixing collected after centrifugation of plasma, -70 low temperature refrigerator frozen. After collecting samples by the method of ELISA Ghrelin detection in plasma Leptin and PYY levels, and compare the results whether there are differences.
3.RT-PCR was used to detect the mRNA expression of Ghrelin, Leptin and PYY in gastric mucosa.
In 43 FD patients (20 cases were Hp positive and 23 Hp negative cases) were included in this study, in the morning of 8~10 were performed with conventional gastroscopy in the gastric antrum mucosa of two blocks, a rapid urease test to detect Hp infection, another piece in the EP tube, -70 low temperature refrigerator to be frozen specimens collect after the detection of Ghrelin RT-PCR method, the expression of Leptin and PYY mRNA, were compared between Hp+ group and Hp- group the result of whether there is a difference.
4.HE staining and immunohistochemistry:
The 71 FD cases, Hp+23 cases, Hp-48 cases (including EPS25 cases, PDS23 cases), 21 cases of healthy children as control group. The gastric mucosa is fixed, sliced, HE stained to select 5 clear different HPF, 400 times microscope statistics the number of eosinophils and summation. The expression of factor Eotaxin and RANTES method to detect the chemotaxis in gastric mucosa, each slice randomly selected 3 representative field, average gray 400 times under light microscope to measure and calculate each section immunohistochemical staining, and compare the results.
Result
1. epidemiological survey results: (1) the investigation of 12923 cases of children diagnosed FD1034 cases, EPS646 cases, PDS388 cases, Suzhou city children's FD total prevalence rate was 8%. EPS for 5%, PDS for 3%. (2) FD 1034 cases of children by 13C breath test and diagnosis of 282 cases were Hp positive Hp infection rate of 27.3%. (3) PDS related factors of prevalence of parents: the younger, lower educational level, caries more slim, the higher the prevalence (4) related factors of EPS prevalence rate of dental caries: female, more, the higher education level of father, lean or overweight, prevalence rate the higher.
The level of Ghrelin 2. in plasma: FD group than in the control group, PDS group than in EPS group, Hp+ group than in Hp- group, the differences were statistically significant (P < 0.001); plasma Leptin level of FD group was higher than that of control group, PDS group than in EPS group, Hp+ group than in Hp- group, the differences were statistically significant (P < 0.001); the level of plasma PYY in FD group was higher than the control group, the difference was statistically significant (P < 0.001), PDS group and EPS group, Hp+ group and Hp- group, the difference was not statistically significant.
3., in gastric mucosa, the level of ghrelin mRNA in group Hp+ was significantly lower than that in group Hp-, and the level of leptinmRNA in group Hp+ was significantly higher than that in group Hp- (P < 0.01), and PYY was hardly expressed in gastric mucosa of children.
4. gastric mucosal eosinophil count, the expression level of Eotaxin and RANTES in Hp+ and EPS/Hp- group was significantly higher than that of PDS/Hp- group and control group, the differences were statistically significant (P < 0.001). The trend of cytoplasmic Eotaxin gene mainly distributed in gastric mucosa epithelial cells and glandular epithelial cells, and inflammatory cells were not found in distribution.RANTES in the gastric mucosa mainly distributed in gastric mucosal epithelial cells, gland cells, cell cytoplasm and nucleus, the expression level of Eotaxin and RANTES were positively associated with the number of eosinophil
conclusion
1.FD is a common disease of children in Suzhou area. The main subtype is EPS. children. The risk factors for FD are female, younger age, parental education level, Hp infection, dental caries and BMI decrease.
2Ghrelin, Leptin and PYY three kinds of gastrointestinal hormones involved in the pathogenesis of functional dyspepsia in children, Ghrelin and Leptin in the postprandial discomfort (PDS) subtypes in more obvious changes, suggesting that PDS early satiety, anorexia and other symptoms have direct relation with Ghrelin and Leptin, Ghrelin may become a new target of drug therapy PDS.
3. eosinophils and chemokines eotaxin and RANTES are mainly involved in the pathogenesis and disease progression of children with functional dyspepsia and upper abdominal pain (EPS), which provide experimental evidence for clinical research and development of new drugs for EPS.

【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R725.7

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