EGF在新生儿坏死性小肠结肠炎中肠道定位和表达特征

doi:10.3969/j.issn.1000-8535.2017.05.004

摘要/Abstract

摘要： 目的观察表皮生长因子(EGF)在新生儿坏死性小肠结肠炎(NEC)患儿肠组织中的动态表达情况,探讨EGF在NEC病程中起到的保护作用。方法选取15例NEC患儿行一期回肠造瘘手术治疗的回肠组织为实验组(NEC组),将以上15例NEC患儿行二期回肠封瘘手术治疗的回肠组织为对照组(封瘘组),采用免疫组织化学技术检测,通过光密度测算软件(IPP)分析回肠组织中的EGF表达。结果 EGF主要表达于肠壁黏膜层,少量表达于黏膜下层、肌层。EGF在NEC组各层表达平均光密度值为:黏膜层(0.241±0.075),黏膜下层(0.213±0.061),肌层(0.1397±0.026),差异有统计学意义(P＜0.05);在封瘘组各层表达情况为:黏膜层(0.211±0.028),黏膜下层(0.119±0.022),肌层(0.097±0.007),差异有统计学意义(P＜0.05)。EGF在NEC组总体表达平均光密度值为(0.198±0.071),明显高于封瘘组(0.146±0.058),差异有统计学意义(P＜0.05)。结论表皮生长因子(EGF)在新生儿坏死性小肠结肠炎(NEC)肠组织中的表达较封瘘组显著上调,推测EGF可能与NEC炎症相关,可能在NEC炎症过程中起到了一定的保护作用。

关键词: 表皮生长因子, 新生儿坏死性小肠结肠炎, 免疫组织化学

Abstract: Objective We realized that EGF could play an important protective role against NEC. However, the practical condition of the distribution and expression of EGF in intestine of infants with NEC was indefinite. In order to figure out this problem,we carried out this experimentation. Methods The sample were divided into two group.The experimental group(necgroup) were composed of fifteen individual intestinal tissues after the ileostomy were performed on those infants suffered from NEC. The control group(sealing fistula group) were composed of fifteen individual intestinal tissues after the ileal closure fistula were performed on the same infants who were accepted the one-stage ileostomy in the period of NEC and were later accepted the two-stage operation on the condition that their bodies almost recovered from NEC after two to three months gone.Then, we utilized immunohistochemistry to test the distribution and quantities of EGF on those samples of the two group infants. Results The characteristic of EGF expression in intestine of the both group included strong positive expression in mucous layer and less expression in strata submucosum and muscular coat. The average optical density in nec group was mucous layer (0.241±0.075),strata submucosum(0.213±0.026),muscular coat (0.1397±0.022);In the control groupmucous layer (0.211±0.028),strata submucosum (0.119±0.022),muscular coat (0.097±0.007). The expression of EGF in intestinal tissues increased in the period of NEC0.198±0.071 by comparing with the control group (0.146±0.058). Conclusion There may be a correlation between the strong positive expression of EGF in intestinal tissues in the period of NEC and inflammation.By combining the result of this experiment and the research about EGF. We assumed that EGF is one factor of the protective mechanism by which injured intestinal mucous could be recovered and resist inflammation.

Key words: Epidermal growth factor, Neonatal necrotizing enterocolitis, Immunohistochemistry

农一敏, 吕俊健, 何秋明, 宋燕燕, 夏慧敏. EGF在新生儿坏死性小肠结肠炎中肠道定位和表达特征[J]. 广州医药, 2017, 48(5): 14-18.

NONG Yimin, LV Junjian, HE Qiuming , SONG Yanyan, XIA Huimin. Expression of epidermal growth factor in the intestinal tissues of those neonatus with neonatal necrotizing enterocolitis[J]. Guangzhou Medical Journal, 2017, 48(5): 14-18.

参考文献

[1] YOST C C. Neonatal necrotizing enterocolitis: diagnosis, management, and pathogenesis[J]. J Infus Nurs, 2005, 28(2): 130-134.
[2] HEINZERLING N P, LIEDEL J L, WELAK S R, et al. Intestinal alkaline phosphatase is protective to the preterm rat pup intestine[J]. J Pediatr Surg, 2014, 49(6): 954-960.
[3] SHARMA R,HUDAK M L. A clinical perspective of necrotizing enterocolitis: past, present, and future [J]. Clin Perinatol, 2013, 40(1): 27-51.
[4] GOOD M, SIGGERS R H, SODHI C P, et al. Amniotic fluid inhibits Toll-like receptor 4 signaling in the fetal and neonatal intestinal epithelium[J]. Proc Natl Acad Sci U S A, 2012, 109(28): 11330-11335.
[5] HALPERN M D, DOMINGUEZ J A, DVORAKOVA K, et al. Ileal cytokine dysregulation in experimental necrotizing enterocolitis is reduced by epidermal growth factor[J]. J Pediatr Gastroenterol Nutr, 2003, 36(1): 126-133.
[6] MAYNARD A A, DVORAK K, KHAILOVA L, et al. Epidermal growth factor reduces autophagy in intestinal epithelium and in the rat model of necrotizing enterocolitis[J]. Am J Physiol Gastrointest Liver Physiol, 2010, 299(3): G614-622.
[7] CLARK J A, LANE R H, MACLENNAN N K, et al. Epidermal growth factor reduces intestinal apoptosis in an experimental model of necrotizing enterocolitis[J]. Ibid, 2005, 288(4): G755-762.
[8] ITO Y, TAKEDA T, SAKON M, et al. Expression and clinical significance of erb-B receptor family in hepatocellular carcinoma[J]. Br J Cancer, 2001, 84(10): 1377-1383.
[9] BARNARD J A, BEAUCHAMP R D, RUSSELL W E, et al. Epidermal growth factor-related peptides and their relevance to gastrointestinal pathophysiology[J]. Gastroenterology, 1995, 108(2): 564-580.
[10] BERSETH C L. Enhancement of intestinal growth in neonatal rats by epidermal growth factor in milk[J]. Am J Physiol, 1987, 253(5 Pt 1): G662-665.
[11] O'LOUGHLIN E V, CHUNG M, HOLLENBERG M, et al. Effect of epidermal growth factor on ontogeny of the gastrointestinal tract[J]. Ibid, 1985, 249(6 Pt 1): G674-678.
[12] OPLETA-MADSEN K, MEDDINGS J B,GALL D G. Epidermal growth factor and postnatal development of intestinal transport and membrane structure[J]. Pediatr Res, 1991, 30(4): 342-350.
[13] MIETTINEN P J, BERGER J E, MENESES J, et al. Epithelial immaturity and multiorgan failure in mice lacking epidermal growth factor receptor[J]. Nature, 1995, 376(6538): 337-341.
[14] AL-NAFUSSI A I,WRIGHT N A. The effect of epidermal growth factor (EGF) on cell proliferation of the gastrointestinal mucosa in rodents[J]. Virchows Arch B Cell Pathol Incl Mol Pathol, 1982, 40(1): 63-69.
[15] HORMI K,LEHY T. Developmental expression of transforming growth factor-alpha and epidermal growth factor receptor proteins in the human pancreas and digestive tract [J]. Cell Tissue Res, 1994, 278(3): 439-450.
[16] JANKOWSKI J, MURPHY S, COGHILL G, et al. Epidermal growth factor receptors in the oesophagus[J]. Gut, 1992, 33(4): 439-443.
[17] SULLIVAN P B, BRUETON M J, TABARA Z B, et al. Epidermal growth factor in necrotising enteritis[J]. Lancet, 1991, 338(8758): 53-54.
[18] MENESHIAN A,BULKLEY G B. The physiology of endothelial xanthine oxidase: from urate catabolism to reperfusion injury to inflammatory signal transduction[J]. Microcirculation, 2002, 9(3): 161-175.
[19] CHAILLER P,MENARD D. Ontogeny of EGF receptors in the human gut[J]. Front Biosci, 1999(4): D87-101.
[20] FAGBEMI A O, WRIGHT N, LAKHOO K, et al. Immunoreactive epidermal growth factor receptors are present in gastrointestinal epithelial cells of preterm infants with necrotising enterocolitis[J]. Early Hum Dev, 2001, 65(1): 1-9.
[21] MENARD D ,POTHIER P. Radioautographic localization of epidermal growth factor receptors in human fetal gut[J]. Gastroenterology, 1991, 101(3): 640-649.
[22] MENARD D, ARSENAULT P ,POTHIER P. Biologic effects of epidermal growth factor in human fetal jejunum[J]. Ibid, 1988(94): 656-663.
[23] MIETTINEN P J, BERGER J E, MENESES J, et al. Epithelial immaturity and multiorgan failure in mice lacking epidermal growth factor receptor[J]. Nature, 1995, 376(6538): 337-341.
[24] SWANIKER F, GUO W, FONKALSRUD E W, et al. The effect of epidermal growth factor on mucosal function after ileal resection[J]. J Surg Res, 1995, 58(6): 565-569.
[25] 宋丹,方秀斌.表皮生长因子的研究进展[J]. 解剖科学进展,2002,8(4):339-342.
[26] AUWERX J. Nuclear receptors. I. PPAR gamma in the gastrointestinal tract: gain or pain? [J]. Am J Physiol Gastrointest Liver Physiol, 2002, 282(4): G581-585.
[27] CLAUD E C, SAVIDGE T,WALKER W A. Modulation of human intestinal epithelial cell IL-8 secretion by human milk factors[J]. Pediatr Res, 2003, 53(3): 419-425.
[28] WALTERS J R. Cell and molecular biology of the small intestine: new insights into differentiation, growth and repair[J]. Curr Opin Gastroenterol, 2004, 20(2): 70-76.
[29] CHANG C J,CHAO J C. Effect of human milk and epidermal growth factor on growth of human intestinal Caco-2 cells[J]. J Pediatr Gastroenterol Nutr, 2002, 34(4): 394-401.