α-萘基异硫氰酸酯诱导的肝损伤对短肠综合征大鼠模型肠道适应性的影响。

Effect of alpha-naphthylisothiocyanate-induced liver injury on intestinal adaptation in a rat model of short bowel syndrome.

作者信息

Sukhotnik Igor, Razon Hila, Pollak Yulia, Hayari Lili, Bejar Jacob, Mogilner Jorge G, Sylvester Karl G

机构信息

Laboratory of Intestinal Adaptation and Recovery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.

出版信息

Pediatr Surg Int. 2012 Feb;28(2):161-9. doi: 10.1007/s00383-011-2989-y.

DOI:10.1007/s00383-011-2989-y

PMID:21989949

Abstract

BACKGROUND/PURPOSE: Progressive hyperbilirubinemia and end-stage liver failure are among the most serious complications of short bowel syndrome (SBS), representing the principle cause of death in a majority of fatal cases. In the current study, we examined the effects of alpha-naphthylisothiocyanate (ANIT)-induced liver injury on intestinal adaptation in a rat model of SBS.

METHODS

Male rats were divided into four groups: Sham rats underwent bowel transection (n = 8), Sham liver-injury rats underwent bowel transection and IP injection of ANIT (100 mg/kg, n = 8), SBS rats underwent a 75% bowel resection, and SBS-ANIT rats underwent bowel resection and liver injury similar to group sham-ANIT (n = 8). Fourteen days after intervention, liver biopsies and intestinal samples were obtained and evaluated for liver damage and measures of intestinal adaptation. Real time PCR and Western blotting were used to determine the level of bax and bcl-2 mRNA and protein, and p-ERK protein levels. Statistical analysis was performed using the one-way ANOVA test, with p < 0.05 considered statistically significant.

RESULTS

All ANIT-treated animals exhibited histological evidence of liver damage that was associated with the expansion of atypical ductal proliferation near the periportal areas, intense neutrophil infiltration in the liver, increased mitotic activity, Kupfer cells hyperplasia and fatty liver degeneration. ANIT-induced liver damage in bowel resected animals was associated with a significant decrease in all parameters of intestinal adaptation including bowel and mucosal weight in jejunum (twofold decrease) and ileum (twofold decrease), mucosal DNA in jejunum (fourfold decrease), mucosal protein in jejunum (threefold decrease) and ileum (threefold decrease), villus height in jejunum (38%) and ileum (34%), and crypt depth in jejunum (24%) and ileum (30%) compared to SBS animals. Both Sham-ANIT and SBS-ANIT rats demonstrated decreased enterocyte proliferation rates that were accompanied by decreased p-ERK protein levels. Lower apoptotic rates in jejunum (40%) and ileum (52%) in SBS-ANIT rats (vs. SBS) coincided with decreased bax mRNA and protein levels.

CONCLUSIONS

In a rat model of SBS, ANIT-induced liver injury was associated with decreased enterocyte proliferation and inhibited intestinal adaptation.

摘要

背景/目的：进行性高胆红素血症和终末期肝衰竭是短肠综合征（SBS）最严重的并发症，是大多数致命病例的主要死因。在本研究中，我们在SBS大鼠模型中研究了α-萘异硫氰酸酯（ANIT）诱导的肝损伤对肠道适应性的影响。

方法

将雄性大鼠分为四组：假手术组大鼠进行肠横断术（n = 8），假手术肝损伤组大鼠进行肠横断术并腹腔注射ANIT（100 mg/kg，n = 8），SBS组大鼠进行75%肠切除术，SBS-ANIT组大鼠进行与假手术-ANIT组相似的肠切除和肝损伤（n = 8）。干预14天后，获取肝脏活检组织和肠道样本，评估肝损伤情况和肠道适应性指标。采用实时PCR和蛋白质印迹法测定bax和bcl-2 mRNA及蛋白水平，以及p-ERK蛋白水平。使用单因素方差分析进行统计分析，p < 0.05被认为具有统计学意义。

结果

所有接受ANIT治疗的动物均表现出肝损伤的组织学证据，这与门周区域附近非典型导管增生的扩大、肝脏中强烈的中性粒细胞浸润、有丝分裂活性增加、库普弗细胞增生和脂肪肝变性有关。在肠切除动物中，ANIT诱导的肝损伤与肠道适应性的所有参数显著降低有关，包括空肠（降低两倍）和回肠（降低两倍）的肠重和黏膜重量、空肠黏膜DNA（降低四倍）、空肠黏膜蛋白（降低三倍）和回肠黏膜蛋白（降低三倍）、空肠绒毛高度（降低38%）和回肠绒毛高度（降低34%），以及空肠隐窝深度（降低24%）和回肠隐窝深度（降低30%），与SBS动物相比。假手术-ANIT组和SBS-ANIT组大鼠的肠上皮细胞增殖率均降低，同时p-ERK蛋白水平降低。SBS-ANIT组大鼠空肠（降低40%）和回肠（降低52%）的凋亡率较低（与SBS组相比），这与bax mRNA和蛋白水平降低一致。