劉寶珍, 屈冬冬, 金世祿, 劉京運(yùn)

(1. 山東省濱州市人民醫(yī)院 消化科; 山東 濱州, 256610;2. 濱州醫(yī)學(xué)院臨床學(xué)院, 山東 濱州, 256603)

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細(xì)胞毒素相關(guān)蛋白A和白細(xì)胞介素-9與消化性潰瘍的關(guān)系研究

劉寶珍1, 屈冬冬2, 金世祿1, 劉京運(yùn)2

(1. 山東省濱州市人民醫(yī)院 消化科; 山東 濱州, 256610;2. 濱州醫(yī)學(xué)院臨床學(xué)院, 山東 濱州, 256603)

目的探討細(xì)胞毒素相關(guān)蛋白A(CagA)、白細(xì)胞介素-9(IL-9)與消化性潰瘍發(fā)病的關(guān)系。方法選取消化性潰瘍患者40例為研究組，慢性胃炎患者40例為對(duì)照組，檢測(cè)2組血清CagA、IL-9水平及幽門螺桿菌(Hp)感染情況等，并對(duì)消化性潰瘍進(jìn)行單因素、多因素Logistic回歸分析。結(jié)果研究組血清CagA水平明顯高于對(duì)照組, IL-9水平明顯低于對(duì)照組(P<0.001); 研究組Hp感染率及CagA+Hp感染率均明顯高于對(duì)照組(P<0.01); CagA+患者血清IL-9水平明顯低于CagA-患者(P<0.01)。Logistic回歸分析顯示, CagA+Hp感染、吸煙、生活不規(guī)律、Ⅰ級(jí)親屬共患病、精神因素、飲酒與消化性潰瘍的發(fā)生密切相關(guān)(P<0.01), CagA+Hp感染與所選分層因素之間在消化性潰瘍的發(fā)生中具有協(xié)同作用。結(jié)論CagA+幽門螺桿菌感染可能與消化性潰瘍的發(fā)生有關(guān), IL-9可能對(duì)Hp感染所致的消化性潰瘍具有抑制作用。消化性潰瘍的發(fā)生是眾多因素共同作用的結(jié)果。

消化性潰瘍; 細(xì)胞毒素相關(guān)蛋白A; 白細(xì)胞介素-9; 幽門螺桿菌

自1983年首次從人胃黏膜活檢標(biāo)本中分離培養(yǎng)出幽門螺桿菌(Hp)后，諸多研究[1-2]表明Hp感染是消化性潰瘍的重要病因之一，與消化性潰瘍的發(fā)生密切相關(guān)，但感染Hp后是否致病取決于不同的毒力株?，F(xiàn)已證實(shí)，一半以上的Hp菌株均表達(dá)細(xì)胞毒素相關(guān)蛋白A(CagA), CagA是Hp的重要毒力因子之一，會(huì)引起更為嚴(yán)重的炎癥反應(yīng)、消化道黏膜損傷等[3-4]。白細(xì)胞介素-9(IL-9)能通過抑制細(xì)胞免疫活性，發(fā)揮抗炎效應(yīng)，具有多效性免疫功能[5]。本研究探討CagA、IL-9與消化性潰瘍發(fā)病的關(guān)系。

1　資料與方法

1.1一般資料

選擇2013年9月—2014年9月就診的消化性潰瘍患者40例為研究組，慢性胃炎患者40例為對(duì)照組，均經(jīng)胃鏡結(jié)合組織病理學(xué)檢查確診，排除患有其他消化道疾病、其他急慢性疾病、惡性腫瘤的患者。其中研究組男30例，女10例；年齡38～66歲，平均(46.5±5.6)歲。對(duì)照組男28例，女12例；年齡37～68歲，平均(47.2±5.3)歲。2組一般資料比較差異無統(tǒng)計(jì)學(xué)意義，具有可比性(P>0.05)。研究方案經(jīng)本院倫理委員會(huì)研究并批準(zhǔn)進(jìn)行，入組患者及家屬均知情同意并書面簽署同意書。

1.2方法

采集所有患者空腹末梢血3 mL, 離心后分離血清，采用ELISA法檢測(cè)各組血清CagA、IL-9水平。根據(jù)14C-UBT、RUT和病理性特殊染色確定Hp陽(yáng)性(至少有2項(xiàng)測(cè)定結(jié)果陽(yáng)性則判定為Hp陽(yáng)性)；對(duì)Hp感染者采用斑點(diǎn)金免疫滲濾法檢測(cè)血清抗Hp-CagA IgG。根據(jù)設(shè)計(jì)假設(shè)，選擇可能與消化性潰瘍發(fā)生有關(guān)的危險(xiǎn)因素并制定統(tǒng)一調(diào)查表[4], 內(nèi)容主要包括個(gè)人資料、生活習(xí)慣、健康狀況、家族史、社會(huì)經(jīng)濟(jì)、衛(wèi)生狀況、精神因素等，對(duì)消化性潰瘍進(jìn)行單因素、多因素Logistic回歸分析。

1.3統(tǒng)計(jì)學(xué)分析

2　結(jié)　果

2.12組血清CagA、IL-9水平及Hp感染情況

研究組血清CagA水平明顯高于對(duì)照組, IL-9水平明顯低于對(duì)照組(P<0.01)。研究組Hp感染率為92.5%(37/40), 明顯高于對(duì)照組的47.5%(19/40)(P<0.01)。見表1。

與對(duì)照組比較, **P<0.01。

2.2血清CagA+HP感染率及IL-9水平比較比較

研究組CagA+HP感染率為90.0%(36/40), 明顯高于對(duì)照組的22.5%(9/40)；研究組、對(duì)照組CagA+Hp血清IL-9水平均明顯低于CagA-Hp患者(P<0.01)。見表2。

表2　2組CagA+比例及其血清IL-9水平比較[n(%)]

與CagA-比較, **P<0.01。

2.3消化性潰瘍單因素條件Logistic回歸分析

Logistic回歸分析顯示, CagA+Hp感染、吸煙、生活不規(guī)律、Ⅰ級(jí)親屬共患病、精神因素、飲酒與消化性潰瘍的發(fā)生密切相關(guān)(P<0.01), 其中CagA+Hp感染與消化性潰瘍的關(guān)系最為密切, OR值最大，生活不規(guī)律次之。見表3。

表3　消化性潰瘍單因素條件Logistic回歸分析

2.4消化性潰瘍多因素條件Logistic回歸分析

對(duì)單因素分析有意義的因素進(jìn)行多元Logistic回歸分析，結(jié)果顯示CagA+Hp感染與所選分層因素之間在消化性潰瘍的發(fā)生中具有協(xié)同作用。見表4。

表4　消化性潰瘍多因素條件Logistic回歸分析

3　討　論

Hp廣泛存在于人體，與消化性潰瘍、胃炎、胃癌等多種消化系統(tǒng)疾病密切相關(guān)[6]。研究[7]認(rèn)為Hp產(chǎn)生的毒素CagA雖然含量甚微，但能通過白細(xì)胞趨化及激活，引起強(qiáng)烈的免疫反應(yīng)和組織損傷。本研究通過ELISA法檢測(cè)患者血清CagA, 結(jié)果顯示研究組血清CagA水平明顯高于對(duì)照組，提示CagA與消化性潰瘍的關(guān)系較慢性胃炎更為密切。CagA基因表達(dá)分子量為120～145 KDa的CagA蛋白，能夠通過PAI構(gòu)成的分泌系統(tǒng)進(jìn)入消化道上皮細(xì)胞，破壞細(xì)胞頂端連接復(fù)合體的結(jié)構(gòu)及功能，干擾上皮細(xì)胞分化，降解基膜。研究[8]認(rèn)為, CagA陽(yáng)性與胃十二指腸疾病有關(guān)。Caliskan等[9]將492例患者分為無癥狀對(duì)照組、慢性萎縮性胃炎組、胃腺癌組、十二指腸球部潰瘍組及胃潰瘍組5組，計(jì)算各組Hp及CagA+Hp的感染率，結(jié)果顯示, Hp相關(guān)性疾病如慢性萎縮性胃炎、胃腺癌及消化性潰瘍時(shí)，以CagA+Hp感染為主, CagA+Hp感染率較無癥狀對(duì)照組更高。近年來，Hp感染與消化性潰瘍的關(guān)系受到學(xué)者們的普遍關(guān)注，諸多研究結(jié)果均證實(shí)Hp感染是消化性潰瘍發(fā)病諸因素中的危險(xiǎn)因素之一。國(guó)外學(xué)者Saber等[10]采用ELSIA法檢測(cè)患者對(duì)CagA抗體的免疫反應(yīng)，發(fā)現(xiàn)超過70%的胃潰瘍患者和100%的十二指腸潰瘍患者呈陽(yáng)性。Cheng等[11]也報(bào)道，十二指腸潰瘍患者血清中的CagA抗體檢出率為84%, 顯著高于非潰瘍性消化道疾病患者。本研究結(jié)果顯示研究組與對(duì)照組Hp感染率分別為92.5%、47.5%, CagA陽(yáng)性HP感染率分別為90.0%、22.5%, 研究組Hp感染率及CagA+HP感染率均明顯高于對(duì)照組。提示CagA陽(yáng)性Hp感染與消化道潰瘍的發(fā)生密切相關(guān)，抗CagA抗體陽(yáng)性的檢測(cè)結(jié)果能夠作為消化性潰瘍的臨床診斷參考指標(biāo)。

IL-9是分子量為14 KDa的糖蛋白，具有雙重調(diào)節(jié)功能，既可負(fù)向調(diào)節(jié)炎癥反應(yīng)、增加T細(xì)胞免疫抑制活性，又可以通過激活肥大細(xì)胞、巨噬細(xì)胞等促過敏反應(yīng)及炎癥反應(yīng)[12]。Defendenti等[13]在實(shí)驗(yàn)研究中構(gòu)建了Hp感染的小鼠模型，結(jié)果顯示Hp感染組血清IL-9水平明顯低于Hp未感染組；進(jìn)一步在臨床上檢測(cè)消化系統(tǒng)疾病患者血清標(biāo)本，結(jié)果發(fā)現(xiàn)Hp陰性的受試者血清IL-9水平明顯高于Hp陽(yáng)性的患者。上述結(jié)果提示, IL-9表達(dá)減少與Hp感染相關(guān)，兩者具有互相抑制的作用。本研究通過檢測(cè)對(duì)2組血清IL-9水平，結(jié)果提示IL-9可能對(duì)消化性潰瘍及慢性胃炎均具有抑制作用。根據(jù)是否存在CagA+Hp感染，進(jìn)一步比較2組間血清IL-9水平，結(jié)果顯示CagA+患者血清IL-9水平均明顯低于CagA-患者，提示CagA可能在Hp抑制IL-9的表達(dá)中發(fā)揮重要作用。

研究[14]認(rèn)為，消化性潰瘍可能是多種因素共同作用的結(jié)果，本研究通過Logistic回歸分析顯示, CagA+Hp感染、吸煙、生活不規(guī)律、Ⅰ級(jí)親屬共患病、精神因素、吸煙與消化性潰瘍的發(fā)生密切相關(guān)，且CagA+Hp感染與所選分層因素之間在消化性潰瘍的發(fā)生中具有協(xié)同作用。

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Study on relationship of cytotoxin associated gene A and interleukin-9 with peptic ulcer

LIU Baozhen1, QU Dongdong2, JIN Shilu1, LIU Jingyun2

(1.DepartmentofGastroenterology,BinzhouPeople′sHospital,Binzhou,Shandong, 256610;2.ClinicalSchool,BinzhouMedicalUniversity,Binzhou,Shandong, 256603)

ObjectiveTo explore the relationship of cytotoxin associated gene A (CagA) and interleukin-9 (IL-9) with peptic ulcer. MethodsA total of 40 patients with peptic ulcer were selected as research group while another 40 patients with chronic gastritis as control group. Serum CagA and IL-9 levels as well as helicobacter pylori (Hp) infection condition were detected in both groups, and univariate and multivariate Logistic regression analysis were conducted to patients with peptic ulcer. ResultsResearch group was markedly higher in serum CagA level but evidently lower in IL-9 level than control group (P<0.001). Research group was notably higher in Hp infection rate and CagA-positive Hp infection rate than control group (P<0.01). Patients with positive CagA+were prominently lower in serum IL-9 level than those with negative CagA-(P<0.01). Logistic regression analysis indicated that CagA+Hp infection, smoking, irregular lifestyle, co-illness with relatives in degree Ⅰ, mental factors and alcoholic consumption were closely associated with the occurrence of peptic ulcer (P<0.01), and CagA+Hp infection had synergistic effect with selected stratification factors in the development of peptic ulcer. ConclusionCagA+Hp infection may be associated with the development of peptic ulcer, and IL-9 has potential inhibitory effect in peptic ulcer induced by Hp infection. And the occurrence of peptic ulcer is in close association with multiple factors.

peptic ulcer; cytotoxin associated gene A; interleukin-9; helicobacter pylori

2016-04-13

中國(guó)高校醫(yī)學(xué)期刊臨床專項(xiàng)資金(11527692)

屈冬冬, E-mail: Qddcool@163.com

R 573.1

A

1672-2353(2016)17-057-03

10.7619/jcmp.201617018