CAJ | 학술논문

目的明确M细胞在急性胰腺炎时细胞因子TNF-α、1L-18的表达情况,探索M细胞在肠黏膜屏障功能中的町能作用.方法 SD大鼠,随机分为急性胰腺炎6、12、24、48 h组及对照组,每组10只,建立大鼠急性胰腺炎模型,收集标本.检测血清淀粉酶、谷丙转氨酶的变化,鲎试剂法检测血清内毒素变化,ELISA法检测血清内细胞因子TNF-α、IL-18的变化,检测肠系膜淋巴结细菌移位情况,观察同肠黏膜病理形态学改变,LCM、RT-PCR法检测M细胞中TNF-α mRNA、IL-18mRNA的表达,免疫荧光双重染色法观察M细胞中TNF-α、IL-18的表达.结果 (1)血淀粉酶在胰腺炎6、12、24 h组升高,48 h组下降.(2)血清内毒素在急性胰腺炎12、24、48 h组升高,6 h组无明显升高,肠系膜淋巴结细菌移位菌落数在急性胰腺炎各时间点均升高.(3)血清TNF-α在急性胰腺炎组6、12、24 h组升高,48 h组下降到正常水平,血清IL-18在胰腺炎各时间点均升高,在24 h达到最高水平.(4)病理检查发现,在急性胰腺炎组随着时间延长回肠黏膜水肿逐渐加重.(5)免疫荧光双重染色结果显示,在急性胰腺炎组的各时间段M细胞TNF-α蛋白表达均为阳性,在6 h、12 h和24 h组M细胞内的IL-18蛋白表达呈明显阳性,对照组阴性.(6)在急性胰腺炎组各时间TNF-αmRNA的表达量均高于对照组.从术后6h开始升高并达到一个平台期,在12 h和24 h维持在较高水平,到术后48 h TNF-αmRNA的表达量较前下降但仍高于对照组.在6 h、12 h IL-18 mRNA的表达升高,48 h下降.结论在急性胰腺炎,肠黏膜屏障功能受到不同程度损害,M细胞合成TNF-α、IL-18增加,M细胞可能在肠道黏膜免疫反应中发挥一定作用,但还需进一步研究.
목적 명학M세포재급성이선염시세포인자TNF-α、1L-18적표체정황,탐색M세포재장점막병장공능중적정능작용.방법 SD대서,수궤분위급성이선염6、12、24、48 h조급대조조,매조10지,건립대서급성이선염모형,수집표본.검측혈청정분매、곡병전안매적변화,후시제법검측혈청내독소변화,ELISA법검측혈청내세포인자TNF-α、IL-18적변화,검측장계막림파결세균이위정황,관찰동장점막병리형태학개변,LCM、RT-PCR법검측M세포중TNF-α mRNA、IL-18mRNA적표체,면역형광쌍중염색법관찰M세포중TNF-α、IL-18적표체.결과 (1)혈정분매재이선염6、12、24 h조승고,48 h조하강.(2)혈청내독소재급성이선염12、24、48 h조승고,6 h조무명현승고,장계막림파결세균이위균락수재급성이선염각시간점균승고.(3)혈청TNF-α재급성이선염조6、12、24 h조승고,48 h조하강도정상수평,혈청IL-18재이선염각시간점균승고,재24 h체도최고수평.(4)병리검사발현,재급성이선염조수착시간연장회장점막수종축점가중.(5)면역형광쌍중염색결과현시,재급성이선염조적각시간단M세포TNF-α단백표체균위양성,재6 h、12 h화24 h조M세포내적IL-18단백표체정명현양성,대조조음성.(6)재급성이선염조각시간TNF-αmRNA적표체량균고우대조조.종술후6h개시승고병체도일개평태기,재12 h화24 h유지재교고수평,도술후48 h TNF-αmRNA적표체량교전하강단잉고우대조조.재6 h、12 h IL-18 mRNA적표체승고,48 h하강.결론 재급성이선염,장점막병장공능수도불동정도손해,M세포합성TNF-α、IL-18증가,M세포가능재장도점막면역반응중발휘일정작용,단환수진일보연구.
Objective To investigate TNF-α and IL-18 expression levels in mierofold cells in rat' s intestine after acute panereatitis to evaluate the role of mierofold cells (M cells) in intestinal barrier.Methods Fifty SD rats were randomized into the acute panereatitis 6 h group,12 h group,24 h group,48 h group and control group.The model of acute pancreatitis was established.Serum,peyer,s patch,ileum,and mesenteric lymph nodes (MLN) were harvested.Serum amylase,glutamate pyrurate transaminse,total bilirubin,endotoxin,TNF-α,and IL-18 were detected.Pathologic changes of the ileal mucous were observed.M cells were obtained with laser capture microdisseetion and the expression of TNF-α mRNA and IL-18 mRNA in M cells were observed with RT-PCR.The expression of TNF-a and IL-18 was determined with immunofluoreseence dual staining.Result 1)The level of serum amylase in pancreatitis increased obviously in 6,12,24 h group,but decreased in 48 h group.2) Endotoxin in blood serum obviously increased in 12,24,48 hours after acute pancreatitis.The number of colony of bacterium aversion in MLN increased in acute pancreatitis.3)TNF-a in blood serum obviously increased in 6,12,24 h groups in acute pancreatitis,and then decreased to normal level in 48 h.IL-18 in blood serum increased in acute pancreatitis.4)Edema was observed in ileal mucus in acute pancreatitis.5)The immunofluoreseence double staining showed that TNF-α and IL-18protein expressed in M cell of all groups of acute pancreatitis but not in control group.6)The level of TNF-αmRNA and IL-18 mRNA expression was significantly higher in acute panereatitis groups than in the control group.Conclusion Intestinal barrier is damaged in acute pancreatitis. M cells can excrete TNF-α and IL 18 in acute pancreatitis and may play important roles in intestinal mucosal immune.