臨床消化病雜本2001年第13卷第4期乙醇對(duì)胃乙醇脫氫酶活性變化的影響R5 A中國醫(yī)科大學(xué)第一臨床附屬醫(yī)院消化內(nèi)科(沈陽10001)王國祥程云友李軍劉事榮犢萋目的:探討乙醇對(duì)胃乙醇脫氫膨( Alcohol dehydrogenase,ADH活性變化的彩響。方法: Wistar大鼠52隨機(jī)分成兩組:模型組32只,對(duì)照組20只。用乙醇直接灌胃的方法建立大鼠模型;用酶組織細(xì)胞化學(xué)染色技術(shù),觀察胃DH活性變化,并用 LUZEX-F灰度圖像分析儀進(jìn)行半定量。結(jié)果:隨砦乙醇灌注量的增加和灌注時(shí)間的延長,胃ADH活性進(jìn)行性下降,與對(duì)照組有顯著性差異(P<0.05);停止癟注后1個(gè)月,胃ADH活性明顯恢復(fù),且與正常對(duì)照組無顯著性差異(P>0.05)。結(jié)論:乙醇導(dǎo)致胃ADH活性進(jìn)行性下降從而降低了乙醇首過代謝,增加了乙醇對(duì)肝臟等器官的毒害作用關(guān)鍵調(diào)乙醇脫氫酶乙醇組織細(xì)胞化學(xué)胃Effect of Alcohol on Active Alteration of Gastric Alcohol DehydrogenaseWang Guo-xiang, Cheng Yun-you, LI Jun, et alDigestion Department of Internal Medicine, The First Armlliated Clinical Haspital, China Medical Uaiversity, Shenyangl1000lAbstract Purpose: Effect of alcohol on activity variety of gastric alcohol dehydrogenase( ADH)wasinvestigated. Methods: Fifty-two rats were divided into two groups in model one( 32/52)and control one(20/52), at random. Model one was made of directly perfusing alcohol into stomach. The activity varietiesof gastrobserved by the optical microscopy after gastric tissues were stained by enzyme histo-cytochemistry assay. The activity varieties were determined by LuzEx-F analysis instrument as a semimethod. Results: It was shown that the activities of gastric adh gradually decreased alongthlume of alcohol increased and the duration of perfusion prolonged. There were significant differences between model group and control one( P <0. 05). The activity of gastric ADH significantly recoveredone month after abstinence from alcohol. There were not significant differences between two groups(P>0. 05). Conclusion: Alcohol resulted in activity descending of gastric ADH, as a result of decreasing firatpass metabolism of gastric alcohol and increasing alcoholic poisonous roles in liver and other organsKey words Alcohol dehydrogenase Alcohol Enzyme histo-cytochemistry assay Stomach乙醇脫氫酶( Alcohol dehydrogenase,ADH)是參下,用平衡飼料喂養(yǎng)。與乙醇代謝的重要酶類之一(。它主要氧化乙醇成1.2實(shí)驗(yàn)方法乙醛,乙醛又在乙醛脫氫酶作用下生成乙酸最后分解1.2.1動(dòng)物模型建立:實(shí)驗(yàn)動(dòng)物隨機(jī)分為兩組,其中成二氧化碳和水。近年來,胃ADH在乙醇首過代謝對(duì)照組20只模型組32只。正常喂養(yǎng)1周后開始建( First pass metabolism,FPM)中的作用倍受人們矚立模型。(1)模型組給予40%的乙醇(國產(chǎn)分析醇),8目認(rèn)為胃ADH在乙醇FPM中起主要作用23。然g/(kgd),分3次灌胃;至第4周末將乙醇濃度升至而目前關(guān)于長期飲酒后的胃ADH活性變化的研究較50%,9g/(kgd,分3次灌胃;至第8周末,乙醇量升少,另外,胃ADH活性的變化與胃粘膜病理改變的關(guān)至10g/(kgd),分3次灌胃;至第12周末模型建立成系亦未確定,為此我們進(jìn)行了相關(guān)的實(shí)驗(yàn)性研究功。(2)對(duì)照組給予相同體積的蒸餾水灌胃,日3次。兩組均于開始灌胃后4周末、8周末、12周末分批處1對(duì)象和方法死模型組各8只,對(duì)照組各5只。兩組余下的動(dòng)物用平衡飼料判1個(gè)目后處研聯(lián)酪后迅速切取新鮮胃1,1實(shí)驗(yàn)對(duì)象6周齡雄性wsar大鼠52只,體重體組織00±10g,由中國醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物部提供,每籠8查。CNMHG只,分籠飼養(yǎng)。按晝夜時(shí)程,在室溫及穩(wěn)定濕度條件1.2.2胃組織病理學(xué)檢查:胃組織行常規(guī)HE染色臨床消化病雜志201年第13第4期159光鏡下觀察胃組織(主要是粘膜層)的病理改變。表2停止灌注乙醇后1個(gè)月模型組與對(duì)照組胃ADH灰度值1.23ADH酶組織化學(xué)檢測方法1:(1)迅速把組織x±s)切成適當(dāng)大小(約5×5×3mm),放入OCT包埋劑,用組別ADH(x±)干冰丙酮或液氮迅速凍結(jié)。(2)用 Cryostat(-1516周對(duì)照組129.09±11.7020℃)制成12m之冰凍切片。(3)用0.1MPBS(p型組130.01±9.104)稀釋2%~4%的戊二醛固定1分鐘。(4)用0.1與對(duì)照組相比P>005MPBs洗45分鐘或過夜。(5)切片在下述孵育液中孵育,用NBT法為37℃,60分鐘。乙醇四唑鹽還原3討論酶的孵育液:0.1MPBS(pH7.4)10m,NAD’60mg,45%乙醇3ml,NBT6ml,蒸餾水1ml,PMS3ADH是乙醇代謝通路上的重要酶類之一,它主mg。(6)用蒸餾水沖洗。(7)用甘油膠封固。(8)對(duì)照要氧化乙醇成乙醛,乙醛又在乙醛脫氫酶作用下生成實(shí)驗(yàn):在孵育液中,不加乙醇或n丁醇而用蒸餾水代乙酸最后代謝成二氧化碳和水。目前已發(fā)現(xiàn)的ADH替。光鏡下通過觀察反應(yīng)產(chǎn)物沉淀密度的變化來反映有7個(gè)基因型表現(xiàn)型分為5類。ADH主要為aBy、各組之間ADH的活性變化;并用 LUZXC-F灰度圖像xx、0亞單位組成二聚體,分布廣泛,主要分布于胃腸分析儀(日本產(chǎn))測定ADH的灰度值進(jìn)行指標(biāo)量化。和肝臟內(nèi)。胃ADH主要包括3類即I類、Ⅲ類Ⅳ類1.3統(tǒng)計(jì)學(xué)處理:結(jié)果經(jīng)計(jì)算機(jī)SAS6.11統(tǒng)計(jì)軟件ADH。在1985年, Julknmen等6發(fā)現(xiàn)靜脈給予和進(jìn)行方差分析和q檢驗(yàn)?？诜葦?shù)量的乙醇后其血乙醇和乙酸水平是不同的。他們發(fā)現(xiàn)靜脈給乙醇組的血乙醇水平明顯高于口2結(jié)果服組的血乙醇水平;而且血乙酸即乙醇代謝產(chǎn)物的水平在口服給予乙醇者比靜脈給予乙醇者明顯升高。由21胃組織病理學(xué)改變光鏡下模型組與對(duì)照組此推論:乙醇部分容量沒有進(jìn)入全身循環(huán),而在胃腸道HE染色結(jié)果對(duì)比可見胃粘膜層細(xì)胞(包括主細(xì)胞和代謝這種部分乙醇在胃腸內(nèi)的代謝叫乙醇首過代謝壁細(xì)胞)水腫變性;且48、12周末模型組主細(xì)胞和壁( First pass metabolism,FPM)細(xì)胞水腫變性逐漸加重。停止灌注乙醇后1月模型組乙醇FPM可調(diào)節(jié)機(jī)體對(duì)乙醇的生物利用度,緩胃粘膜層細(xì)胞(包括主細(xì)胞和壁細(xì)胞)恢復(fù)正常。解乙醇對(duì)肝臟、腦及機(jī)體其它器官的毒害作用。因此2.2胃組織ADH酶組織細(xì)胞化學(xué)染色結(jié)果胃FPM的研究引起了學(xué)術(shù)界相當(dāng)大的關(guān)注。DpadADH主要定位于周粘膜層的壁細(xì)胞和主細(xì)胞,呈紫藍(lán)等報(bào)道FPM在餐后比在禁食情況下大得多;FPM色。停止灌注乙醇前各模型組胃ADH活性明顯低于在女性要比男性低。 Caballeria等3!報(bào)道胃切除病人對(duì)照組(P<001),并隨乙醇灌注量的增加和灌注時(shí)比健康人低。近年來胃ADH在乙醇的FPM中作用間的延長4周8周12周胃粘膜層ADH的活性進(jìn)行倍受人們矚目認(rèn)為胃ADH在乙醇的FM中起主要性下降(P<0.05)。停止灌注乙醇后1個(gè)月模型組胃作用t2S。粘膜ADH活性明顯恢復(fù)(P>0.05)。酶組織細(xì)胞化在本實(shí)驗(yàn)中,我們應(yīng)用酶組化方法研究長期灌注學(xué)染色 LUZEX-F灰度值結(jié)果見表1、2。乙醇后大鼠的胃ADH活性變化及停止灌注乙醇后的1停止灌注乙醇前攘型組與對(duì)照組胃粘胰ADH灰度值胃ADH活性改變,同時(shí)進(jìn)行胃ADH的細(xì)胞定位,及x±)胃ADH活性變化與胃粘膜病理變化的關(guān)系。研究發(fā)現(xiàn)模型組的胃ADH活性較對(duì)照組的胃ADH活性明4周對(duì)照組5127110顯下降并隨乙醇灌注量的增加和灌注時(shí)間的延長呈模型組138,21±8.86進(jìn)行性下降,而停止灌注乙醇后1月胃ADH活性恢8周對(duì)照組復(fù)正常。這與 watanabe9的報(bào)告一致。提示長期飲換型組15847±11.96酒會(huì)降12周對(duì)照組126.31±11.71從而增中國煤化工度,加重了乙糗型組8:17021:1.0。醇對(duì)機(jī)fCNMHGH反應(yīng)產(chǎn)物主與對(duì)照組相比P<001,與前期相比P<0.05要沉淀于粘膜層的壁細(xì)胞和主細(xì)胞的胞漿中,說明了方數(shù)據(jù)臨床消化病雜志2001年第13巷第4期胃ADH主要位于壁細(xì)胞和主細(xì)胞的胞漿中。995,216:216另外,我們對(duì)停止灌注乙醇前后大鼠的胃粘膜進(jìn)3chle,FaM. Hemander-manozr,tra. Gasric origin行了同步HE染色,發(fā)現(xiàn)模型組4周、8周、12周末胃the first-pass metabolism of ethanol in humans: effect of gastrectomyGastroenterology, 1989, 97:1205枯膜層璧細(xì)胞和主細(xì)胞發(fā)生水腫變性,且隨乙醇灌注4 Toshimitsu W,nkmr. Double staining procedure for histochem量的增加和灌注時(shí)間的延長其程度逐漸加重;而對(duì)照a localization of alcohol and aldehyde dehydrogenase activities in組和停止灌注乙醇后1個(gè)月的模型組胃粘膜均正常。mouse liver. Acta Histochem Cytochem, 1989, 22(3):402這一組織學(xué)結(jié)果提示長期飲酒引起胃粘膜ADH活性5kuhA,ErgwGold bery Genetic markers of alohol abuse.下降可能是由于壁細(xì)胞和主細(xì)胞水腫變性引起的。這Clinical Chimica ActH, 1997, 257: 1996 Jullunen RK, Pipadova C. Lieber CS與Tery提出的乙醇對(duì)胃粘膜損害機(jī)制相一致。ethanol: A gastrointestinal barrier Against the systemic toxicity of由于胃ADH在乙醇的FPM中起重要作用,胃粘thano Life sci, 1985, 37:56膜ADH活性下降導(dǎo)致FPM的下降,從而加大了慢性7wa C, Wormer TM, Julkunen RJ, et al. Effects of fasting and嗜酒者的乙醇生物利用度,加重了乙醇對(duì)肝臟等器官chronic acold consumption on the firan pam metabolism of ethand的負(fù)荷,加速了酒精性肝病等疾病的發(fā)生發(fā)展。因此Gastroenterology, 1987, 92:1169了解冒粘膜ADH活性變化誘導(dǎo)ADH的活性,將為8DdwC.FemM, Lieber Cs. Gastric metabolism of ethandbility in men and women. In: Biomedical酒精性肝病等疾病的預(yù)防和治療提供新的途徑and social aspects of alcohol and alcoholism, ed. Kuriyama K and Ishi魯考文獻(xiàn)1 Shih JY, Chin SL, Chew WW, et al. Human stomach alcohol and 9 WatAnabe M. Immunohistochemical lcalization of ADH in the stomaldehyde dehydrogenases: comparison ol expression pattern and activi-ach before and after abstinence of alcohol in aloobolics-using confocales in alimentary tract. Gastroenterology, 1997, 112: 766laser scanning microscopy. Kurume Med J. 1997, 44(4): 263.Hirokazu yokoyama, Enrique barona, Charles Lierber. Upstream 10 Terry L, Macmath MD. Aloobol and gastrointestinal bleeding. Emergon of its exMed Cin North Am, 1990, 8(4):859pression. Biochemical and Biophysical Research Communication(20010114收20010411修回)缺血性結(jié)腸炎8例診治體會(huì)R55 B贏樊鐵路醫(yī)院(襄樊441003)田新社張慧松襄市第一人民醫(yī)院趙文剛本組缺血性結(jié)腸炎8例,男5例,女3例,年齡58-73歲,平均62歲。其中高血壓病4例次(下同),腦血栓后遺癥1例冠心病1例,糖尿病3例,習(xí)慣性便秘2例慢性支氣管炎、肺氣腫2例。全部病例均有突發(fā)性腹痛。其中便血6例發(fā)熱5例腹脹3例悉心、嘔吐2例。纖維結(jié)腸鏡檢查結(jié)果:乙狀結(jié)腸粘膜充血、水腫、糜爛3例結(jié)腸脾曲粘膜充血、水腫散在淺潰瘍2例降結(jié)腸粘膜增厚、腸腔狹窄1例,乙狀結(jié)腸粘膜充血、水腫、激瘍1例,橫結(jié)腸近牌曲處粘膜散在淺潰瘍1例。病變腸段和粘膜主要表現(xiàn)為腸粘膜輕度彌漫性充血、水腫或散在片狀充血、紅森及意合期潰瘍,血管網(wǎng)仍較模幗不清。病理檢查結(jié)果:均為結(jié)腸粘膜非特異性炎癥。后期復(fù)查可見枯膜下含鐵血黃素沉著。根據(jù)臨床表現(xiàn)纖維結(jié)腸鏡檢查和病理檢查結(jié)果結(jié)合基礎(chǔ)疾病并排除上消化道出血瘍性結(jié)腸炎、結(jié)腸克隆氏病腸結(jié)核、大腸癌、慢性結(jié)腸炎等疾病而確定診斷。治療:(1)擴(kuò)管治療:靜脈點(diǎn)滴復(fù)方丹參、黃芪等,口服消心痛、腸溶阿斯匹林等7~14天。停用縮血管藥物。(2)全身應(yīng)用廣諧抗生素,以預(yù)防和減輕腸道繼發(fā)性細(xì)菌感染,減少細(xì)菌毒米產(chǎn)生,減輕內(nèi)毒素血癥。(3)下消化道出血明顯者于結(jié)腸鏡檢查時(shí)給予凝血酶1000-2000u局部噴灑止血,另外口服凝血酶5000-100u,tid,口殿思密達(dá)等腸粘胰保護(hù)劑。結(jié)果:8例經(jīng)纖維結(jié)腸鏡檢查、病理檢查確診為非壞疽型患者,全部痊意。缺血性結(jié)腸炎可分為壞疽型和非壞疽型兩型。壞疽型為不可逆性腸管缺血、壞死,全身中毒癥狀重有腹膜炎體征,死亡率高應(yīng)盡早剖腹手術(shù),切除病變腸段(20001222收稿中國煤化工CNMHG