McLaughlan J M, Seth R, Vautier G, Robins R A, Scott B B, Hawkey C J, Jenkins D
Department of Clinical Laboratory Sciences, University Hospital, Nottingham, U.K.
J Pathol. 1997 Jan;181(1):87-92. doi: 10.1002/(SICI)1096-9896(199701)181:1<87::AID-PATH736>3.0.CO;2-J.
Interleukin-8 (IL-8) and nitric oxide (NO) may be important mediators in the pathogenesis of chronic idiopathic inflammatory bowel disease (CIIBD), but their roles in disease activity in ulcerative colitis (UC) and Crohn's disease (CD) are uncertain. The aim of this study was to measure mRNA for IL-8 and inducible NO synthase (iNOS) in small mucosal biopsies from untreated patients at first presentation and to relate these measurements to the histological levels of polymorph infiltration graded on a ten-point scale. For this purpose, a sensitive enzyme-linked oligonucleotide chemiluminescent assay (ELOCA) was developed to quantitate reverse transcription-polymerase chain reaction (RT-PCR) products amplified from RNA from paired biopsy samples. The levels of IL-8 and iNOS mRNAs were calculated as ratios of the RT-PCR products to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) RT-PCR product. In UC patients, median values of IL-8/GAPDH and iNOS/GAPDH were significantly elevated compared with controls and CD. However, in both UC and CD, the IL-8/GAPDH and iNOS/GAPDH ratios correlated significantly with polymorph infiltration. ELOCA enabled quantitation of multiple mRNAs in small mucosal biopsies from untreated patients with CIIBD and supported a role for IL-8 and iNOS in acute inflammation in both UC and CD.
白细胞介素-8(IL-8)和一氧化氮(NO)可能是慢性特发性炎症性肠病(CIIBD)发病机制中的重要介质,但它们在溃疡性结肠炎(UC)和克罗恩病(CD)疾病活动中的作用尚不确定。本研究的目的是测量初诊时未经治疗患者的小肠黏膜活检组织中IL-8和诱导型一氧化氮合酶(iNOS)的mRNA,并将这些测量结果与按十分制分级的多形核白细胞浸润的组织学水平相关联。为此,开发了一种灵敏的酶联寡核苷酸化学发光测定法(ELOCA),以定量从配对活检样本的RNA中扩增的逆转录-聚合酶链反应(RT-PCR)产物。IL-8和iNOS mRNA的水平计算为RT-PCR产物与甘油醛-3-磷酸脱氢酶(GAPDH)RT-PCR产物的比值。与对照组和CD患者相比,UC患者中IL-8/GAPDH和iNOS/GAPDH的中位数显著升高。然而,在UC和CD患者中,IL-8/GAPDH和iNOS/GAPDH比值均与多形核白细胞浸润显著相关。ELOCA能够定量未经治疗的CIIBD患者小肠黏膜活检组织中的多种mRNA,并支持IL-8和iNOS在UC和CD急性炎症中的作用。