布拉氏酵母菌对NEC新生鼠肠组织中NOD1、RIP2和NF-κB表达的影响
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摘要
目的探讨布拉氏酵母菌(SB)对坏死性小肠结肠炎(NEC)新生小鼠肠组织中的NOD1、受体相互作用蛋白2(RIP2)和核因子-κB(NF-κB)表达的影响及意义。方法 40只新生小鼠随机分为4组,每组10只,分为正常对照组(母乳喂养不予其他干预)、人工喂养组(仅予人工喂养)、NEC组(人工喂养+缺氧复氧冷刺激+脂多糖灌胃)及NEC+SB组在NEC组基础上加SB干预,800mg/kg·d,灌胃1次/d,连用3d。HE染色和病理评分来评估回盲部肠组织的损伤程度; Western blot和RT-qPCR技术分别检测结肠组织NOD1、RIP2和NF-κB的蛋白和mRNA表达水平。结果人工喂养组新生鼠回盲部肠组织病理评分较正常对照组略微升高,NEC组病理评分较正常组显著升高,而NEC+SB组病理评分较NEC组显著下降;人工喂养组和NEC组新生鼠结肠组织中NOD1、RIP2和NF-κB的蛋白和mRNA水平较正常对照组均升高,NEC+SB组NOD1、RIP2和NF-κB蛋白和mRNA表达水平较人工喂养组和NEC组均显著降低,差异有统计学意义(P<0. 05);使用Ad-NOD1(携带NOD1基因的腺病毒)过表达NOD1后,病理评分增加,NOD1、RIP2和NF-κB蛋白表达显著增加,差异有统计学意义(P<0. 01)。结论 SB可能通过抑制NEC新生鼠肠组织中NOD1炎症信号通路相关因子(NOD1、RIP2和NF-κB)的表达水平,从而减轻NEC导致的肠组织损伤。
Objective To investigate the effect of saccharomyces boulardii( SB) on the expression of NOD1( nucleotide oligomerization domain 1),receptor interacting protein 2( RIP2) and nuclear factor-κB( NF-κB) in neonatal necrotizing enterocolitis( NEC) mice. Methods 40 newborn mice were randomly divided into 4 groups( 10 in each group) : control group,formula feeding group( formula feeding only),NEC group( formula feeding+hypoxia-reoxygen treatment+clod exposure+lipopolysaccharide intragastric administration),and NEC+SB group[NEC group plus SB intervention,800 mg/( kg·d),gavage once/d,3 d]. HE staining and pathological scoring were used to evaluate the degree of injury in the ileocecal intestinal tissue. Western blot and RT-qPCR were used to detect the protein and mRNA expression levels of NOD1,RIP2 and NF-κB in colon tissue. Results In formula feeding group,the intestinal pathological score of the ileocecal area of the newborn rats was slightly higher than that of normal control group. The pathological score of NEC group was significantly increased compared with normal group,while the pathological score of NEC+SB group was dramatically decreased compared with NEC group. In addition,the protein and mRNA levels of NOD1,RIP2 and NF-κB in neonatal mice colon tissue of formula feeding and NEC groups were higher than those in control group,and the protein and mRNA levrls of NOD1,RIP2 and NF-κB in NEC+SB group were significantly higher than those in formula feeding group and NEC group( P<0. 05). Using the Ad-NOD1 to overexpress the NOD1,the pathological score increased and the protein level of NOD1、RIP2 and NF-κB increased( P< 0. 01). Conclusion SB may attenuate neonatal mice intestinal tissue damage caused by NEC by inhibiting the expression levels of NOD1 inflammatory signaling pathway-related factors( NOD1,RIP2 and NF-κB).
Objective To investigate the effect of saccharomyces boulardii( SB) on the expression of NOD1( nucleotide oligomerization domain 1),receptor interacting protein 2( RIP2) and nuclear factor-κB( NF-κB) in neonatal necrotizing enterocolitis( NEC) mice. Methods 40 newborn mice were randomly divided into 4 groups( 10 in each group) : control group,formula feeding group( formula feeding only),NEC group( formula feeding+hypoxia-reoxygen treatment+clod exposure+lipopolysaccharide intragastric administration),and NEC+SB group[NEC group plus SB intervention,800 mg/( kg·d),gavage once/d,3 d]. HE staining and pathological scoring were used to evaluate the degree of injury in the ileocecal intestinal tissue. Western blot and RT-qPCR were used to detect the protein and mRNA expression levels of NOD1,RIP2 and NF-κB in colon tissue. Results In formula feeding group,the intestinal pathological score of the ileocecal area of the newborn rats was slightly higher than that of normal control group. The pathological score of NEC group was significantly increased compared with normal group,while the pathological score of NEC+SB group was dramatically decreased compared with NEC group. In addition,the protein and mRNA levels of NOD1,RIP2 and NF-κB in neonatal mice colon tissue of formula feeding and NEC groups were higher than those in control group,and the protein and mRNA levrls of NOD1,RIP2 and NF-κB in NEC+SB group were significantly higher than those in formula feeding group and NEC group( P<0. 05). Using the Ad-NOD1 to overexpress the NOD1,the pathological score increased and the protein level of NOD1、RIP2 and NF-κB increased( P< 0. 01). Conclusion SB may attenuate neonatal mice intestinal tissue damage caused by NEC by inhibiting the expression levels of NOD1 inflammatory signaling pathway-related factors( NOD1,RIP2 and NF-κB).
引文
[1]Augusto Zani,Agostino Pierro.Necrotizing enterocolitis:controversies and challenges[J].F1000Res,2015,4.
[2]Alfaleh K,Anabrees J,Bassler D,et al.Probiotics for prevention of necrotizing enterocolitis in preterm infants[J].Evidence-Based Child Health,2015,5(1):339-368.
[3]陈凌,陈煜,黄萍.布拉氏酵母菌对不同喂养方式极低出生体重儿坏死性小肠结肠炎的影响研究[J].齐齐哈尔医学院学报,2015,36(30):4533-4536.
[4]Akisu M,Baka M,Yalaz M,et al.Supplementation with Saccharomyces boulardiia meliorates hypoxia/reoxygenation induced necrotizing enterocolitis in young mice[J].European joumal of pediatric surgery:official journal of Austrian Association of Pediatric Surgery,2003,13(5):319-323.
[5]María-Alexandra Ca1as,María-JoséFábrega,Rosa Giménez,et al.Outer Membrane Vesicles From Probiotic and Commensal Escherichia coli Activate NOD1-Mediated Immune Responses in Intestinal Epithelial Cells[J].Front Microbiol.2018,9:498.
[6]SophieMayle,Joseph P.Boyle,Eiki Sekine,et al.Engagement of Nucleotide-binding Oligomerization Domain-containing Protein 1(NOD1)by Receptor-interacting Protein 2(RIP2)Is Insufficient for Signal Transduction[J].J Biol Chem,2014,289(33):22900-22914.
[7]Barna J,Renner E,Arszovszki A,et al.Suckling induced activation pattern in the brain of rat pups[J].Nutritional Neuroscience,2017,21(5):317-327.
[8]Premkumar MH,Sule G,Nagamani SC,et al.Argininosuccinate lyase in enterocytes protects from development of necrotizing enterocolitis[J].Am J Physiol Gastrointest Liver Physiol,2014,307(3):347-354.
[9]Nadler EP,Dickinson E,Knisely A,et al.Expression of inducible nitric oxide synthase and interleukin-12 in experimental necrotizing enterocolitis[J].J Surg Res,2000,92(1):71-77.
[10]Hung-Yang Chang,Jin-Hua Chen,Jui-Hsing Chang,et al.Multiple strains probiotics appear to be the most effective probiotics in the prevention of necrotizing enterocolitis and mortality:An updated meta-analysis[J].PLoS One,2017,12(2):e0171579.
[11]Wu Y,Zhong XY,Jiang J,et al.Prospective and controlled study on effect of fortified human milk feeding on infants with extremelyand very low birth weight during hospital stay[J].Journal of Peking University,2016,48(1):143-148.
[12]Fernandez-Carrocera LA,Solis-Herera A,Cabanillas-Ayon M,et al.Double-blind randomised clinical assay to evaluate the eficacy of probiotics in preterm new borns weighing less than 1500 g in the prevention of necrotising enterocolitis[J].Arch Dis Child Fetal Neonatal Ed,2013,98(1):5-9.
[13]Mark A.Underwood.Impact of Probiotics on Necrotizing Enterocolitis[J].Semin Perinatol,2017,41(1):41-51.
[14]GladysVillarruel,Daniel Martinez Rubio,Fani Lopez,et al.Saccharomyces boulardii in acute childhood diarrhea:a randomized,placebocontrolled study[J].Acta Pdiatr,2007,96(4):538-541.
[15]Clarke TB,Weiser JN.Intracellular sensors of extracellular bacteria[J].Immunol Rev,2011,243:9-25.
[16]Philpott DJ,Girardin SE.Nod-like receptors:sentinels at host membranes[J].Curr Opin Immunol,2010,22:428-434.
[17]Roberta Caruso,Neil Warner,Naohiro Inohara,et al.NOD1 and NOD2:Signaling,Host Defense,and Inflammatory Disease[J].Immunity,2014,41(6):898-908.
[18]Chamaillard M,Hashimoto M,Horie Y,et al.An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid[J].Nat Immunol,2003,4:702-707.
[19]Michael H.Shaw,Thornik Reimer,Yun-Gi Kim,et al.OD-like Receptors(NLRs):Bona Fide Intracellular Microbial Sensors[J].Curr Opin Immunol.2008,20(4):377-382.
[20]Jitendra Maharana,Sukanta Kumar Pradhan and Sachinandan De.NOD1CARDMight Be Using Multiple Interfaces for RIP2-Mediated CARD-CARD Interaction:Insights from Molecular Dynamics Simulation[J].PLoS One.2017,12(1):e0170232.
[21]Woon Geon Shin,Bum Joon Park,Sung Joong Lee,et al.Infection of human intestinal epithelial cells by invasive bacteria activates NF-κBand increases ICAM-1 expression through NOD1[J].Korean J Intern Med,2018,33(1):81-90.
[2]Alfaleh K,Anabrees J,Bassler D,et al.Probiotics for prevention of necrotizing enterocolitis in preterm infants[J].Evidence-Based Child Health,2015,5(1):339-368.
[3]陈凌,陈煜,黄萍.布拉氏酵母菌对不同喂养方式极低出生体重儿坏死性小肠结肠炎的影响研究[J].齐齐哈尔医学院学报,2015,36(30):4533-4536.
[4]Akisu M,Baka M,Yalaz M,et al.Supplementation with Saccharomyces boulardiia meliorates hypoxia/reoxygenation induced necrotizing enterocolitis in young mice[J].European joumal of pediatric surgery:official journal of Austrian Association of Pediatric Surgery,2003,13(5):319-323.
[5]María-Alexandra Ca1as,María-JoséFábrega,Rosa Giménez,et al.Outer Membrane Vesicles From Probiotic and Commensal Escherichia coli Activate NOD1-Mediated Immune Responses in Intestinal Epithelial Cells[J].Front Microbiol.2018,9:498.
[6]SophieMayle,Joseph P.Boyle,Eiki Sekine,et al.Engagement of Nucleotide-binding Oligomerization Domain-containing Protein 1(NOD1)by Receptor-interacting Protein 2(RIP2)Is Insufficient for Signal Transduction[J].J Biol Chem,2014,289(33):22900-22914.
[7]Barna J,Renner E,Arszovszki A,et al.Suckling induced activation pattern in the brain of rat pups[J].Nutritional Neuroscience,2017,21(5):317-327.
[8]Premkumar MH,Sule G,Nagamani SC,et al.Argininosuccinate lyase in enterocytes protects from development of necrotizing enterocolitis[J].Am J Physiol Gastrointest Liver Physiol,2014,307(3):347-354.
[9]Nadler EP,Dickinson E,Knisely A,et al.Expression of inducible nitric oxide synthase and interleukin-12 in experimental necrotizing enterocolitis[J].J Surg Res,2000,92(1):71-77.
[10]Hung-Yang Chang,Jin-Hua Chen,Jui-Hsing Chang,et al.Multiple strains probiotics appear to be the most effective probiotics in the prevention of necrotizing enterocolitis and mortality:An updated meta-analysis[J].PLoS One,2017,12(2):e0171579.
[11]Wu Y,Zhong XY,Jiang J,et al.Prospective and controlled study on effect of fortified human milk feeding on infants with extremelyand very low birth weight during hospital stay[J].Journal of Peking University,2016,48(1):143-148.
[12]Fernandez-Carrocera LA,Solis-Herera A,Cabanillas-Ayon M,et al.Double-blind randomised clinical assay to evaluate the eficacy of probiotics in preterm new borns weighing less than 1500 g in the prevention of necrotising enterocolitis[J].Arch Dis Child Fetal Neonatal Ed,2013,98(1):5-9.
[13]Mark A.Underwood.Impact of Probiotics on Necrotizing Enterocolitis[J].Semin Perinatol,2017,41(1):41-51.
[14]GladysVillarruel,Daniel Martinez Rubio,Fani Lopez,et al.Saccharomyces boulardii in acute childhood diarrhea:a randomized,placebocontrolled study[J].Acta Pdiatr,2007,96(4):538-541.
[15]Clarke TB,Weiser JN.Intracellular sensors of extracellular bacteria[J].Immunol Rev,2011,243:9-25.
[16]Philpott DJ,Girardin SE.Nod-like receptors:sentinels at host membranes[J].Curr Opin Immunol,2010,22:428-434.
[17]Roberta Caruso,Neil Warner,Naohiro Inohara,et al.NOD1 and NOD2:Signaling,Host Defense,and Inflammatory Disease[J].Immunity,2014,41(6):898-908.
[18]Chamaillard M,Hashimoto M,Horie Y,et al.An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid[J].Nat Immunol,2003,4:702-707.
[19]Michael H.Shaw,Thornik Reimer,Yun-Gi Kim,et al.OD-like Receptors(NLRs):Bona Fide Intracellular Microbial Sensors[J].Curr Opin Immunol.2008,20(4):377-382.
[20]Jitendra Maharana,Sukanta Kumar Pradhan and Sachinandan De.NOD1CARDMight Be Using Multiple Interfaces for RIP2-Mediated CARD-CARD Interaction:Insights from Molecular Dynamics Simulation[J].PLoS One.2017,12(1):e0170232.
[21]Woon Geon Shin,Bum Joon Park,Sung Joong Lee,et al.Infection of human intestinal epithelial cells by invasive bacteria activates NF-κBand increases ICAM-1 expression through NOD1[J].Korean J Intern Med,2018,33(1):81-90.
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