消化性潰瘍患者外周血細(xì)胞免疫及體液免疫功能變化分析

吳 蓉，李國熊，李 麗，周 剛，吳建良，陳 晶

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消化性潰瘍患者外周血細(xì)胞免疫及體液免疫功能變化分析

吳 蓉，李國熊，李 麗，周 剛，吳建良，陳 晶

310015浙江省杭州市，杭州師范大學(xué)附屬醫(yī)院消化內(nèi)科

【摘要】目的探討消化性潰瘍患者外周血細(xì)胞免疫及體液免疫功能的變化及臨床意義。方法選擇2014年1月—2015年1月杭州師范大學(xué)附屬醫(yī)院門診及住院的連續(xù)經(jīng)胃鏡確診的消化性潰瘍患者122例為潰瘍組，另選擇同期體檢中心體檢健康者70例為對照組，潰瘍組患者根據(jù)是否感染幽門螺桿菌分為幽門螺桿菌陽性亞組(Hp+亞組)85例和幽門螺桿菌陰性亞組(Hp-亞組)37例，根據(jù)是否出血分為出血亞組58例和未出血亞組64例。檢測外周血細(xì)胞免疫指標(biāo)(包括CD3、CD4、CD8、CD4/CD8、CD19、CD56)及體液免疫指標(biāo)(IgG、IgA、IgM、IgE、C3、C4)水平，出血亞組中30例大便隱血試驗(yàn)陽性患者大便隱血試驗(yàn)轉(zhuǎn)陰后復(fù)查外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)。結(jié)果潰瘍組患者外周血CD3、CD4、CD4/CD8、CD19、CD56、C3、C4水平較對照組降低，IgG水平較對照組升高(P<0.05)；Hp+亞組患者外周血CD3、CD4、CD8、CD19、IgM、IgE水平較Hp-亞組升高，CD4/CD8較Hp-亞組降低(P<0.05)；出血亞組患者外周血CD3、CD4水平、CD4/CD8較未出血亞組降低(P<0.05)。大便隱血試驗(yàn)陽性患者轉(zhuǎn)陰后外周血CD3、CD4、CD8、CD19、CD56、C3、C4水平較治療前升高，IgG、IgM水平較治療前降低(P<0.05)。結(jié)論消化性潰瘍患者存在免疫功能調(diào)節(jié)紊亂，幽門螺桿菌感染可促使機(jī)體產(chǎn)生強(qiáng)烈的免疫反應(yīng)，急性出血后免疫功能處于抑制狀態(tài)，止血后免疫功能有所恢復(fù)。

【關(guān)鍵詞】消化性潰瘍；幽門螺桿菌；消化性潰瘍出血；細(xì)胞免疫；體液免疫

吳蓉，李國熊，李麗，等.消化性潰瘍患者外周血細(xì)胞免疫及體液免疫功能變化分析[J].中國全科醫(yī)學(xué)，2016，19(20)：2481-2485.[www.chinagp.net]

WU R,LI G X,LI L,et al.Changes of cellular immune function and humoral immune function in peripheral blood of patients with peptic ulcer[J].Chinese General Practice，2016，19(20)：2481-2485.

消化性潰瘍是一種多發(fā)病、常見病，好發(fā)于胃和十二指腸。免疫因素是否參與了消化性潰瘍的發(fā)生與發(fā)展，是目前國內(nèi)外研究的焦點(diǎn)之一[1-2]，且一直存在爭議，免疫功能調(diào)節(jié)具有雙向性，其亢進(jìn)或低下與消化性潰瘍的關(guān)系尚不清楚[3]。為此，本研究檢測消化性潰瘍患者外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)，觀察消化性潰瘍患者與體檢健康者、是否感染幽門螺桿菌及是否出血與免疫功能指標(biāo)的關(guān)系以及出血恢復(fù)期免疫功能指標(biāo)的變化，探討消化性潰瘍與細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)之間的關(guān)系。

1資料與方法

1.1臨床資料選擇2014年1月—2015年1月杭州師范大學(xué)附屬醫(yī)院門診及住院的連續(xù)經(jīng)胃鏡確診的消化性潰瘍患者122例為潰瘍組，其中男85例，女37例；年齡23～90歲，平均年齡(53.1±16.8)歲；胃潰瘍50例，十二指腸球部潰瘍52例，復(fù)合性潰瘍20例；病程1個(gè)月～3年，平均病程(1.1±0.8)年。排除肝硬化或伴食管胃底靜脈曲張破裂出血、門靜脈高壓性胃病出血、惡性腫瘤、自身免疫性疾病、高血壓、糖尿病、冠心病以及變態(tài)反應(yīng)性疾病者。另選擇同期體檢中心體檢健康者70例為對照組，其中男50例，女20例；年齡21～86歲，平均年齡(52.4±18.1)歲。對照組與潰瘍組性別、年齡、體質(zhì)指數(shù)、吸煙率、飲酒率比較，差異均無統(tǒng)計(jì)學(xué)意義(P>0.05，見表1)。本研究獲得杭州師范大學(xué)附屬醫(yī)院醫(yī)學(xué)倫理委員會批準(zhǔn)，受試者均簽署知情同意書。

表1對照組與潰瘍組一般資料比較

Table 1Comparison of general data between control group and ulcer group

組別例數(shù)性別(男/女)年齡(歲)體質(zhì)指數(shù)(kg/m2)吸煙〔n(%)〕飲酒〔n(%)〕對照組7050/2052.4±18.123.3±5.634(48.6)37(52.9)潰瘍組12285/3753.1±16.822.4±4.860(49.2)58(47.5)χ2(t)值0.066-1.051a0.374a0.0070.503P值0.7980.3220.7230.9350.478

注：a為t值；吸煙定義為：每天吸煙不少于1支，持續(xù)1年，或總量不少于18包/年；飲酒定義為：每周飲酒不少于2次，白酒不少于50 g/次或者其他酒類不少于500 ml/次

1.2分組潰瘍組患者根據(jù)是否感染幽門螺桿菌分為幽門螺桿菌陽性亞組(Hp+亞組)85例和幽門螺桿菌陰性亞組(Hp-亞組)37例，幽門螺桿菌測定采用胃黏膜組織切片蘇木素-伊紅(HE)染色法和14C呼氣試驗(yàn)，其中一項(xiàng)陽性即為幽門螺桿菌感染；根據(jù)是否出血分為出血亞組58例和未出血亞組64例，入院前有嘔血和/或排柏油樣大便，嘔吐物或大便隱血試驗(yàn)陽性判斷為出血。Hp+亞組與Hp-亞組患者性別、年齡、體質(zhì)指數(shù)、吸煙率、飲酒率、疾病種類、病程比較，差異均無統(tǒng)計(jì)學(xué)意義(P>0.05，見表2)。出血亞組與未出血亞組患者性別、年齡、體質(zhì)指數(shù)、吸煙率、飲酒率、疾病種類、病程比較，差異均無統(tǒng)計(jì)學(xué)意義(P>0.05，見表3)。

1.3檢測方法清晨空腹抽取潰瘍組患者及對照組受試者外周靜脈血2 ml加入乙二胺四乙酸二鉀(EDTA-K2)抗凝管中，充分混勻后進(jìn)行細(xì)胞免疫指標(biāo)檢測，同時(shí)抽取4 ml全血，3 500 r/min離心10 min(離心半徑20.8 cm)，分離血清，進(jìn)行體液免疫指標(biāo)檢測。使用FACS Clibur 流式細(xì)胞儀(美國BD公司)和Cell Quest軟件測定受試者外周血細(xì)胞免疫指標(biāo)〔包括T淋巴細(xì)胞(CD3)、輔助性T細(xì)胞(CD4)、抑制性T細(xì)胞(CD8)、CD4/CD8、B淋巴細(xì)胞(CD19)、自然殺傷細(xì)胞(CD56)〕水平。采用Immage雙光徑濁度分析儀(美國Beckman-Coulter公司)測定血清體液免疫指標(biāo)(包括免疫球蛋白IgG、IgA、IgM、IgE，補(bǔ)體C3、C4)水平。出血亞組中30例大便隱血試驗(yàn)陽性患者大便隱血試驗(yàn)轉(zhuǎn)陰后復(fù)查外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)。

1.4治療患者均予抑制胃酸、補(bǔ)液治療，合并大出血休克患者輸注紅細(xì)胞懸液200～400 ml。Hp+亞組患者予抗幽門螺桿菌四聯(lián)療法，使用泮托拉唑鈉腸溶膠囊(杭州中美華東制藥有限公司，藥品生產(chǎn)批號131104)40 mg口服，2次/d；膠體果膠鉍膠囊(浙江得恩德制藥有限公司，藥品生產(chǎn)批號20131017)0.2 g口服，3次/d；克拉霉素分散片(江蘇揚(yáng)子江藥業(yè)集團(tuán)有限公司，藥品生產(chǎn)批號1406041)0.5 g口服，2次/d；阿莫西林膠囊(香港澳美制藥廠，藥品生產(chǎn)批號19961)1.0 g口服，2次/d。青霉素過敏者予呋喃唑酮片(天津力生制藥股份有限公司，藥品生產(chǎn)批號1404009)0.1 g口服，2次/d替代治療2周，停用抗生素，4周后復(fù)查胃鏡及14C呼氣試驗(yàn)。

2結(jié)果

2.1對照組與潰瘍組外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)比較潰瘍組外周血CD3、CD4、CD4/CD8、CD19、CD56、C3、C4水平較對照組降低，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；潰瘍組外周血IgG水平較對照組升高，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；對照組與潰瘍組外周血CD8、IgA、IgM、IgE水平比較，差異無統(tǒng)計(jì)學(xué)意義(P>0.05，見表4)。

2.2Hp+亞組與Hp-亞組患者外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)比較Hp+亞組患者外周血CD3、CD4、CD8、CD19、IgM、IgE水平較Hp-亞組升高，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；Hp+亞組患者外周血CD4/CD8較Hp-亞組降低，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；Hp+亞組與Hp-亞組外周血CD56、IgG、IgA、C3、C4水平比較，差異無統(tǒng)計(jì)學(xué)意義(P>0.05，見表5)。

2.3出血亞組與未出血亞組患者外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)比較出血亞組患者外周血CD3、CD4水平、CD4/CD8較未出血亞組降低，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；出血亞組與未出血亞組外周血CD8、CD19、CD56、IgG、IgA、IgM、IgE、C3、C4水平比較，差異無統(tǒng)計(jì)學(xué)意義(P>0.05，見表6)。

2.4大便隱血試驗(yàn)陽性患者治療前及轉(zhuǎn)陰后外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)比較大便隱血試驗(yàn)陽性患者轉(zhuǎn)陰后外周血CD3、CD4、CD8、CD19、CD56、C3、C4水平較治療前升高，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；轉(zhuǎn)陰后外周血IgG、IgM水平較治療前降低，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；治療前與轉(zhuǎn)陰后外周血CD4/CD8、IgA、IgE水平比較，差異無統(tǒng)計(jì)學(xué)意義(P>0.05，見表7)。

3討論

淋巴細(xì)胞在免疫應(yīng)答中起核心作用，T淋巴細(xì)胞亞群是構(gòu)成機(jī)體免疫防御系統(tǒng)的重要細(xì)胞，免疫球蛋白及補(bǔ)體與淋巴細(xì)胞同樣是機(jī)體免疫防御機(jī)制的重要組成部分，在體液免疫中起著重要作用。目前消化性潰瘍病因尚未完全明了，較為明確的病因是幽門螺桿菌感染，幽門螺桿菌感染是否使機(jī)體免疫功能紊亂，導(dǎo)致消化性潰瘍的發(fā)生，目前的研究結(jié)果并不一致[1-3]。

表2　Hp+亞組與Hp-亞組患者一般資料比較

注：a為t值；Hp+亞組為幽門螺桿菌陽性亞組，Hp-亞組為幽門螺桿菌陰性亞組

表3　出血亞組與未出血亞組患者一般資料比較

注：a為t值

表4　對照組與潰瘍組外周血細(xì)胞免疫指標(biāo)及體液免疫指標(biāo)比較( ±s)

Table 5Comparison of the indexes of cellular immune function and humoral immune function in peripheral blood between Hp+ subgroup and Hp- subgroup

組別例數(shù)CD3(×106/L)CD4(×106/L)CD8(×106/L)CD4/CD8CD19(×106/L)CD56(×106/L)IgG(g/L)IgA(g/L)IgM(g/L)IgE(U/ml)C3(g/L)C4(g/L)Hp+亞組851159±518727±356433±2711.91±0.78233±123211±13011.91±2.822.36±0.791.21±0.77193±730.91±0.220.22±0.07Hp-亞組37780±323534±220255±1482.55±1.40168±83196±10912.90±3.942.50±1.050.93±0.46175±820.84±0.200.21±0.07t值5.2824.0105.123-2.8944.1020.852-1.362-0.8142.5921.9311.4300.718P值<0.001<0.001<0.0010.006<0.0010.3920.1620.4030.0140.0480.1550.474

Table 6Comparison of the indexes of cellular immune function and humoral immune function in peripheral blood between bleeding subgroup and non-bleeding subgroup

組別例數(shù)CD3(×106/L)CD4(×106/L)CD8(×106/L)CD4/CD8CD19(×106/L)CD56(×106/L)IgG(g/L)IgA(g/L)IgM(g/L)IgE(U/ml)C3(g/L)C4(g/L)出血亞組58848±515532±361314±1811.85±0.85181±108173±9612.21±3.502.42±1.081.07±0.68275±1280.84±0.220.22±0.06未出血亞組641127±473730±291396±2982.35±1.19221±120226±13712.18±3.082.35±0.701.12±0.70146±940.91±0.200.21±0.07t值-2.316-2.453-1.402-2.017-1.446-1.9050.0490.328-0.2851.477-1.303-0.523P值0.0240.0170.1660.0480.1530.0610.9610.7440.7770.1460.1970.602

Table 7Comparison of the indexes of cellular immune function and humoral immune function in peripheral blood of patients with positive fecal occult blood testing results between before treatment and after their testing results turned negative

組別例數(shù)CD3(×106/L)CD4(×106/L)CD8(×106/L)CD4/CD8CD19(×106/L)CD56(×106/L)IgG(g/L)IgA(g/L)IgM(g/L)IgE(U/ml)C3(g/L)C4(g/L)治療前30486±180312±55174±901.50±0.7690±31139±1610.60±0.422.33±0.750.70±0.13144±300.63±0.130.15±0.05轉(zhuǎn)陰后30537±153350±94186±931.54±0.5699±52171±2210.13±0.672.32±0.870.67±0.12143±410.66±0.160.16±0.06t值-9.089-7.543-13.342-1.543-3.231-15.67214.620-1.38927.698-1.731-8.080-5.392P值<0.001<0.001<0.0010.161<0.001<0.001<0.0010.185<0.0010.092<0.001<0.001

綜上所述，本研究結(jié)果說明免疫功能參與了消化性潰瘍的發(fā)生、發(fā)展，臨床可通過調(diào)節(jié)免疫功能達(dá)到治愈和預(yù)防消化性潰瘍的目的。關(guān)于免疫功能紊亂與消化性潰瘍之間的關(guān)系，尚待大樣本研究進(jìn)一步證實(shí)。

作者貢獻(xiàn)：吳蓉進(jìn)行課題設(shè)計(jì)與實(shí)施、資料收集整理、撰寫論文、成文并對文章負(fù)責(zé)；李麗、周剛、吳建良、陳晶進(jìn)行課題實(shí)施、評估、資料收集；李國熊進(jìn)行質(zhì)量控制及審校。

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(本文編輯：陳素芳)

Changes of Cellular Immune Function and Humoral Immune Function in Peripheral Blood of Patients With Peptic Ulcer

WURong,LIGuo-xiong,LILi,etal.

DepartmentofGastroenterology,theAffiliatedHospitalofHangzhouNormalUniversity,Hangzhou310015,China

【Abstract】ObjectiveTo investigate the changes of cellular immune function and humoral immune function in peripheral blood of patients with peptic ulcer and the clinical significance.MethodsFrom January 2014 to January 2015,we enrolled 122 patients who were definitely diagnosed as peptic ulcer in the Affiliated Hospital of Hangzhou Normal University as ulcer group,and enrolled 70 healthy people who received physical examination as control group.According to whether infection of helicobacter pylori occurred,the patients in ulcer group were divided into helicobacter pylori positive subgroup(Hp+subgroup)(n=85) and helicobacter pylori negative subgroup(Hp-subgroup)(n=37),and were divided into bleeding subgroup(n=58) and non-bleeding subgroup(n=64) according to whether bleeding occurred.The indexes of cellular immune function(including CD3，CD4，CD8，CD4/CD8，CD19and CD56) and indexes of humoral immune function(including IgG，IgA，IgM，IgE，C3 and C4) in peripheral blood were detected.In bleeding subgroup,30 patints who had positive results in fecal occult blood testing received reexamination of the indexes of immune function and humoral immune function in peripheral blood after fecal occult blood testing showed negative results.ResultsThe levels of CD3，CD4，CD4/CD8，CD19，CD56，C3 and C4 in peripheral blood were lower in ulcer group than those in control group,the levels of IgG in peripheral blood was higher in ulcer group than that in control group(P<0.05)；the levels of CD3，CD4，CD8，CD19，IgM and IgE in peripheral blood were higher in Hp+ subgroup than those in Hp- subgroup，the CD4/CD8 in peripheral blood was lower in Hp+ subgroup than that in Hp- subgroup(P<0.05)；the levels of CD3，CD4 and CD4/CD8 in peripheral blood were lower in bleeding subgroup than those in non-bleeding subgroup(P<0.05).The levels of CD3，CD4，CD8，CD19，CD56，C3 and C4 in peripheral blood were higher in patients who had positive results in fecal occult blood testing first but later turned negative than those before treatment(P<0.05);the levels of IgG,IgM in peripheral blood were lower in patients who had positive results in fecal occult blood testing first but later turned negative than those before treatment(P<0.05).ConclusionPatients with peptic ulcer have disorder in immune function,and infection of helicobacter pylori may induce strong immune reactions.Acute hemorrhage may cause a inhibitory state of immune function which may be relieved after hemostasis.

【Key words】Peptic ulcer；Helicobacter pylori；Peptic ulcer hemorrhage；Cellular immunity;Humoral immunity

通信作者:李國熊，310015浙江省杭州市，杭州師范大學(xué)附屬醫(yī)院消化內(nèi)科；E-mail：guoxiongli849@hotmail.com

【中圖分類號】R 573.1

【文獻(xiàn)標(biāo)識碼】B

DOI：10.3969/j.issn.1007-9572.2016.20.025

(收稿日期：2015-11-12；修回日期：2016-03-20)

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