基于相对和绝对定量的蛋白质组学分析的等压标签揭示了孕激素受体、炎症和纤维化在慢传输型便秘中的作用。

Isobaric tags for relative and absolute quantification-based proteomic analysis that reveals the roles of progesterone receptor, inflammation, and fibrosis for slow-transit constipation.

机构信息

Department of Colorectal Surgery, Tianjin Union Medical Center, Tianjin, China.

Department of Pathology, Tianjin Union Medical Center, Tianjin, China.

出版信息

J Gastroenterol Hepatol. 2018 Feb;33(2):385-392. doi: 10.1111/jgh.13873.

DOI:10.1111/jgh.13873

PMID:28699285

Abstract

BACKGROUND AND AIM

Progesterone receptor, inflammation, neurotransmitter expression, and fibrosis are involved in slow-transit constipation. The aim of the present study was to examine whether patients with slow-transit constipation have an overexpression of progesterone receptor and serotonin, which may impair the fibrosis of muscularis propria in colorectal wall.

METHODS

High-resolution colon manometry was used to record the colorectal peristaltic contractions of the proximal ascending and sigmoid colon in patients. Protein samples prepared from frozen sigmoid colon tissue and the proximal margin of the ascending colon of four female patients were compared using isobaric tags for relative and absolute quantification labeling technique coupled to 2D liquid chromatography-tandem mass spectrometry analysis. Immunohistochemical staining of progesterone receptor, serotonin, and fibronectin was performed in paraffin-embedded sigmoid colon tissues and the proximal margin of the ascending colon or ileum from 43 patients with slow-transit constipation.

RESULTS

Among these differentially regulated proteins based on isobaric tags for relative and absolute quantification and liquid chromatography-tandem mass spectrometry analysis, 56 proteins involved in the response to progesterone, inflammation, matrix remodeling, fibrosis, and muscle metabolism. Immunohistochemical staining confirmed that there was significantly higher expression of progesterone receptor (t = 19.19, P = 0.000) and serotonin (t = 13.52, P = 0.004) in sigmoid colon than in the proximal margin of the ascending colon and ileum. Progesterone receptor and fibronectin expression in the outer layer of muscularis propria were higher than in the middle layer.

CONCLUSIONS

These results demonstrate that progesterone receptor, along with inflammation and fibrosis, may take part in slow-transit constipation development.

摘要

背景与目的

孕激素受体、炎症、神经递质表达和纤维化均与慢传输型便秘相关。本研究旨在探究慢传输型便秘患者是否存在孕激素受体和 5-羟色胺表达过度，这可能会损害结直肠壁固有肌层的纤维化。

方法

通过高分辨率结肠测压法记录 4 例女性患者近端升结肠和乙状结肠的结肠蠕动收缩。采用相对和绝对定量标记(isobaric tags for relative and absolute quantification，iTRAQ)联合二维液相色谱-串联质谱分析比较冷冻乙状结肠组织和近端升结肠组织的蛋白样本。对 43 例慢传输型便秘患者的乙状结肠组织石蜡切片和近端升结肠或回肠组织进行孕激素受体、5-羟色胺和纤维连接蛋白的免疫组织化学染色。

结果

基于 iTRAQ 和液相色谱-串联质谱分析的差异调节蛋白中，有 56 种蛋白参与孕激素反应、炎症、基质重塑、纤维化和肌肉代谢。免疫组织化学染色证实，与近端升结肠和回肠相比，乙状结肠组织中孕激素受体（t=19.19，P=0.000）和 5-羟色胺（t=13.52，P=0.004）的表达显著升高。固有肌层外层的孕激素受体和纤维连接蛋白表达高于中层。