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用于评估肠道细胞因子特异性mRNA的黏膜活检的充分性。正常和炎症肠道活检组织及分离细胞中逆转录聚合酶链反应的比较研究。

Adequacy of mucosal biopsies for evaluation of intestinal cytokine-specific mRNA. Comparative study of RT-PCR in biopsies and isolated cells from normal and inflamed intestine.

作者信息

Fukushima K, West G, Fiocchi C

机构信息

Division of Gastroenterology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4952, USA.

出版信息

Dig Dis Sci. 1995 Jul;40(7):1498-505. doi: 10.1007/BF02285198.

DOI:10.1007/BF02285198
PMID:7543040
Abstract

Endoscopic biopsies are being increasingly utilized to investigate cytokine profiles in normal and diseased intestine. To evaluate the adequacy of mucosal biopsies as a source of cytokine-specific mRNA, we measured their content of interleukin-1 beta (IL-1 beta) and interleukin-2 (IL-2) mRNA by reverse transcription-polymerase chain reaction and compared it to that of autologous lamina propria mononuclear cells in control and inflammatory bowel disease-involved specimens. High-quality total RNA was recovered more consistently from cell isolates than from biopsies. Interleukin-1 beta mRNA was reliably detected in both cell and biopsy samples, whereas IL-2 mRNA was measurable in all lamina propria cells but not always in biopsies. Compared to controls, levels of IL-1 beta were significantly elevated in Crohn's disease and ulcerative colitis cells and biopsies, and a weak but significant correlation existed between values derived from the two sources. In contrast, only ulcerative colitis cell isolates but not biopsies contained significantly reduced concentrations of IL-2 mRNA compared to those of control and Crohn's disease samples, and no correlation was found between cell and biopsy contents. Widely different levels of cytokine-specific mRNA were present in closely adjacent biopsies, particularly for IL-2. These results suggest that mucosal biopsies are suitable to assess some but not all cytokine-specific mRNA due to variation in RNA recovery, intrinsic level of cytokine gene expression, and substantial variability of cytokine transcripts.

摘要

内镜活检越来越多地用于研究正常和患病肠道中的细胞因子谱。为了评估黏膜活检作为细胞因子特异性mRNA来源的充分性,我们通过逆转录-聚合酶链反应测量了其白细胞介素-1β(IL-1β)和白细胞介素-2(IL-2)mRNA的含量,并将其与对照和炎症性肠病相关标本中自体固有层单核细胞的含量进行比较。从细胞分离物中比从活检标本中更一致地回收高质量的总RNA。在细胞和活检样本中均可靠地检测到白细胞介素-1βmRNA,而IL-2mRNA在所有固有层细胞中均可测量,但在活检标本中并非总是如此。与对照组相比,克罗恩病和溃疡性结肠炎细胞及活检标本中IL-1β水平显著升高,且两种来源的值之间存在微弱但显著的相关性。相比之下,与对照和克罗恩病样本相比,仅溃疡性结肠炎细胞分离物而非活检标本中IL-2mRNA浓度显著降低,且细胞和活检标本含量之间未发现相关性。在紧邻的活检标本中存在差异很大的细胞因子特异性mRNA水平,尤其是对于IL-2。这些结果表明,由于RNA回收率、细胞因子基因表达的内在水平以及细胞因子转录本的显著变异性,黏膜活检适合评估某些但不是所有细胞因子特异性mRNA。

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