肽YY可减弱肿瘤坏死因子-α诱导的胰腺炎中的转录因子活性。

Peptide YY attenuates transcription factor activity in tumor necrosis factor-alpha-induced pancreatitis.

作者信息

Vona-Davis Linda, Yu Alice, Magabo Kristine, Evans Tracy, Jackson Barbara, Riggs Dale, McFadden David

机构信息

Department of Surgery, West Virginia University, Morgantown, WV 26506, USA.

出版信息

J Am Coll Surg. 2004 Jul;199(1):87-95. doi: 10.1016/j.jamcollsurg.2004.02.008.

DOI:10.1016/j.jamcollsurg.2004.02.008

PMID:15217635

Abstract

BACKGROUND

Acute pancreatitis (AP) is a disease characterized by inflammation. Nuclear factor (NF)-kappaB, Smad proteins, and the steroid hormone family peroxisome proliferator-activated receptors (PPARs) are involved in regulation of gene transcription during the disease process. Peptide YY (PYY), a gastrointestinal hormone, inhibits NF-kappaB translocation to acinar nuclei in tumor necrosis factor (TNF)-alpha-induced AP. We investigated TNF-alpha induction of Smad proteins, PPARalpha/gamma, and NF-kappaB by TNF-alpha, and hypothesized that PYY would attenuate this effect.

STUDY DESIGN

Rat acinar cells were treated with recombinant TNF-alpha (200 ng/mL). PYY (3 to 36) was added at 500 pM at 30 minutes after TNF-alpha treatment until cell harvest at 2 hours. Western blot analysis and intracellular staining of the p65 subunit of NF-kappaB were performed. NF-kappaB, Smad3/4, and PPARalpha/gamma binding activities were determined by protein/DNA array analysis and verified by electrophoretic-mobility shift assay and densitometry.

RESULTS

Cellular localization of NF-kappaB p65 showed nuclear staining within 2 hours, with controls stained in the cytoplasm. With PYY, p65 stained in the cytoplasm. Nuclear p65 was increased significantly (p < 0.05) by TNF-alpha at 2 hours and PYY reduced it. Array analysis revealed upregulation of NF-kappaB, PPARalpha/gamma, and Smad3/4 with TNF-alpha. TNF-alpha stimulated NF-kappaB activation sevenfold, and binding was enhanced (p < 0.05). PYY reduced NF-kappaB binding to control levels. PPAR binding increased 51% after TNF-alpha treatment and was reduced to 33% with PYY. Smad3/4 binding was increased (p < 0.05) above controls with TNF-alpha and PYY reduced it by 40%.

CONCLUSIONS

TNF-alpha increases early nuclear translocation of the p65 subunit of NF-kappaB in acinar cells. Exposure to TNF-alpha activates transcription factors NF-kappaB, Smad3/4, and PPARalpha/gamma. PYY reduces this activation. Treatment with PYY may have therapeutic potential in improving AP.

摘要

背景

急性胰腺炎（AP）是一种以炎症为特征的疾病。核因子（NF）-κB、Smad蛋白以及类固醇激素家族过氧化物酶体增殖物激活受体（PPARs）参与了该疾病过程中的基因转录调控。肽YY（PYY）是一种胃肠激素，可抑制肿瘤坏死因子（TNF）-α诱导的急性胰腺炎中NF-κB向腺泡细胞核的转位。我们研究了TNF-α对Smad蛋白、PPARα/γ和NF-κB的诱导作用，并假设PYY会减弱这种作用。

研究设计

用重组TNF-α（200 ng/mL）处理大鼠腺泡细胞。在TNF-α处理30分钟后，以500 pM的浓度加入PYY（3 - 36），直至2小时后收获细胞。进行NF-κB p65亚基的蛋白质印迹分析和细胞内染色。通过蛋白质/DNA阵列分析测定NF-κB、Smad3/4和PPARα/γ的结合活性，并通过电泳迁移率变动分析和光密度测定进行验证。

结果

NF-κB p65的细胞定位显示，2小时内核染色，而对照组在细胞质中染色。使用PYY时，p65在细胞质中染色。2小时时，TNF-α使核p65显著增加（p < 0.05），而PYY使其降低。阵列分析显示，TNF-α使NF-κB、PPARα/γ和Smad3/4上调。TNF-α刺激NF-κB激活7倍，结合增强（p < 0.05）。PYY将NF-κB结合降低至对照水平。TNF-α处理后PPAR结合增加51%，而使用PYY后降至33%。TNF-α使Smad3/4结合高于对照水平增加（p < 0.05），而PYY使其降低40%。