目的：探讨肠三叶因子对内毒素血症幼鼠肠组织Toll样受体（TLR）2、4表达的调节及对肠组织损伤的影响。方法：24只10日龄Wistar幼鼠随机分为正常对照组（NS组，n=8，生理盐水1 mL/kg腹腔注射）；内毒素血症组（LPS组，n=8，LPS 5 mg/kg 腹腔注射）；肠三叶因子组（ITF组，n=8，重组肠三叶因子 0.1 mL/只＋LPS 5 mg/kg腹腔注射）。于腹腔注射后3 h处死，留取远端回肠组织观察肠组织病理改变，RT-PCR检测肠组织TLR2、4-mRNA的表达。结果：光镜下NS组肠组织结构正常，ITF组和LPS组均可见间质和上皮细胞水肿，ITF组较LPS组病变明显减轻。ITF组肠组织TLR2 mRNA 表达较NS组、LPS组明显增高（P＜0.01）； 而TLR4 mRNA表达较NS组和LPS组明显下降（P＜0.01）。结论：肠三叶因子可减轻内毒素血症幼鼠肠组织损伤，这种保护作用可能与其下调TLR4 mRNA的表达相关。
Abstract
OBJECTIVE: To study the effects of intestinal trefoil factor (ITF) on Toll-like receptors (TLR) 2 and 4 expression in intestinal tissue and on intestinal injury in young rats with endotoxemia. METHODS: A total of 24 10-day-old Wistar rat pups were equally randomly divided into three groups: a control group, intraperitoneally injected with normal saline 1 mL/kg; an endotoxemia group, intraperitoneally injected with lipopolysaccharide (LPS) 5 mg/kg and an ITF group,intraperitoneally injected with rITF 0.1 mL/each plus LPS 5 mg/kg. Rats were sacrificed 3 h after injection. A segment of distal ileum was dissected for pathologic examinations under an optical microscope (hematoxylin-eosin staning). The mRNA expressions of TLR2 and TLR4 were detected by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: The structure of the small intestine remained normal in the control group. Edema of interstitial substance and epithelium were observed in both LPS and ITF groups, whereas such changes were significantly lower in the ITF group than in the LPS group. The ITF group had significantly higher TLR2 mRNA expression than the NS and LPS groups (P＜0.01), whereas the mRNA expression of TLR4 in the ITF group was significantly lower than in the NS and LPS groups (both P＜0.01). CONCLUSIONS: ITF can alleviate intestinal injury in young rats with endotoxemia, which may be related to the down-regulation of TLR4 mRNA expression.
关键词
肠三叶因子 /
TLR /
肠组织 /
大鼠
Key words
Intestinal trefoil factor /
Toll-like receptor /
Intestinal tissue /
Rats
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
［1］陈美娅, 张 靖, 任建林. 三叶因子3研究进展［J］. 世界华人消化杂志，2008， 16(20): 2267-2273.
［2］Li J, Zhou R, He WC, Xia B. Effects of recombinant human intestinal trefoil factor on trinitrobenzene sulphonic acid induced colitis in rats［J］. Mol Biol Rep, 2011 38(7):4787-4792.
［3］Uematsu S, Akira S. Toll-like receptors and innate immunity［J］. J Mol Med, 2006, 84(9): 712-725.
［4］Peterson CY, Costantini TW, Loomis WH, Putnam JG, Wolf P, Bansal V, et al. Toll-like receptor-4 mediates intestinal barrier breakdown after thermal injury［J］. Surg Infect (Larchmt), 2010, 11(2): 137-144.
［5］de Foneska A, Kaunitz JD. Gastroduodenal mucosal defense［J］. Curr Opin Gastroenterol, 2010, 26(6): 604-610.
［6］Lorenz E.TLR2 and TLR4 expression during bacterial infections［J］. Curr Pharm Des, 2006, 12(32): 4185-4193.
［7］吴秀清，王虹，孙梅，吕庆杰，周卓. 幼年大鼠内毒素血症时小肠上皮细胞凋亡及Caspase-3的表达［J］. 中国当代儿科杂志, 2005, 7(2), 167-170.
［8］李军，许玲芬，孙梅，李强，高红，姜卫国. 肠三叶因子在内毒素致幼鼠肠损伤中的作用及意义［J］. 中国当代儿科杂志, 2006, 8(5):425-428.
［9］John LJ, Fromm M, Schulzke JD. Epithelial barriers in intestinal inflammation［J］. Antioxid Redox Signal, 2011, 15(5):1255-1270.
［10］李刚，李幼生，黎介寿. 肠功能障碍的新概念: 急性肠损伤与急性肠伤害综合征［J］. 肠外与肠内营养, 2010,17(5),302-305.
［11］孙勇, 彭曦. 肠三叶因子研究进展［J］. 肠外与肠内营养,2006，13（4）：243-246.
［12］周雅芮,张丙宏,严彩霞,张海霞,赵日红. 肠三叶因子对新生鼠坏死性小肠结肠炎模型 Caspase29表达的影响［J］. 实用儿科临床杂志，2009, 24(19):1490-1495.
［13］滕旭，许玲芬，孙梅，吴捷，刘璐. 肠三叶因子对炎症性肠病小鼠细胞凋亡的调节机制［J］. 实用儿科临床杂志，2010，26(13):1028-1031.
［14］付春花, 张丙宏, 严彩霞, 陈丽萍, 麦根荣. 肠三叶因子对新生大鼠坏死性小肠结肠炎的保护作用［J］. 中国当代儿科杂志, 2005, 7(1):20-24.
［15］Renes IB, Verburg M, Van Nispen DJ, Büller HA, Dekker J, Einerhand AW. Distinct epitherial responses in-experimental colitis: implications for ion uptake mucosal protection［J］. Am J Physiol Gastrointest Liver Physiol, 2002, 283(1):G169-G179.
［16］Dunzendorfer S, Lee HK, Soldau K, Tobias PS. Toll-like receptor 4 functions intracellularly in human coronary artery endothelial cells: roles of LBP and sCD14 in mediating LPS responses［J］. FASEB J, 2004, 18(10): 1117-1119.
［17］Ohnishi T, Muroi M, Tanamoto K.The lipopolysaccharide-recognition mechanism in cells expressing TLR4 and CD14 but lacking MD-2［J］. FEMS Immunol Med Microbiol, 2007, 51(1): 84-91.
［18］Ortega-Cava CF, Ishihara S, Rumi MA, Kawashima K, Ishimura N, Kazumori H, et al. Strategic compartmentalization of Toll-like receptor 4 in the mouse gut［J］. J Immunol, 2003, 170(8): 3977-3985.
［19］Uematsu S, Fujimoto K.The innate immune system in the intestine［J］. Microbiol Immunol. 2010,54(11):645-657.
［20］Miyake K.Innate recognition of lipopolysaccharide by CD14 and toll-like receptor 4-MD-2: unique roles for MD-2［J］. Int Immunopharmacol, 2003, 3(1): 119-128.
［21］Miyake K.Endotoxin recognition molecules, Toll-like receptor 4-MD-2［J］. Semin Immunol, 2004, 16(1): 11-16.
PDF(1338 KB)
