双歧杆菌对坏死性小肠结肠炎新生小鼠肠组织的保护作用  

Protective effect of Bifidobacterium on endothelial nitric oxide synthase in neonatal necrotizing enterocolitis model

作　　者：邓沁 郭春宝[1] Deng Qin;Guo Chun-Bao(Department of Hepaobiliary Surgery,Children’s Hospital of Chongqing Medical University/Ministry of Education Key Laboratory of Children Development and Disorders/China International Science and Technology Cooperation Base of Child Development and Critical Disorders/Chongqing Key Laboratory of Pediatric,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院肝胆外科/儿童发育疾病研究教育部重点实验室/儿童发育重大疾病国家国际合作基地/儿科学重庆市重点实验室

出　　处：《解放军医学杂志》2020年第1期49-54,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(30973440,30770950);国家自然科学基金重点项目(30330590);重庆市自然科学基金重点项目(CSTC,2008BA0021,cstc2012jj A0155)~~

摘　　要：目的探讨双歧杆菌对坏死性小肠结肠炎(NEC)新生小鼠肠组织的保护作用。方法60只新生8日龄C57BL/6小鼠随机分为对照组、坏死性小肠结肠炎(NEC)组、双歧杆菌干预(NEC+BIF)组,每组20只,建模3 d后处死。HE染色观察新生小鼠肠组织病理形态学变化;Western blotting检测新生小鼠肠组织内皮型一氧化氮合酶(eNOS)蛋白表达情况;比色法检测肠组织eNOS活性及一氧化氮(NO)、超氧阴离子(O2•)含量;实时荧光定量PCR检测肠组织白介素6(IL-6)、白介素1β(IL-1β)的mRNA表达情况;ELISA检测肠组织IL-6、IL-1β蛋白表达情况。结果3组新生小鼠肠组织eNOS蛋白表达差异无统计学意义(P>0.05);与对照组相比,NEC组新生小鼠肠组织eNOS活性降低,NO含量减少,O2^•含量增加,IL-6、IL-1β的mRNA及蛋白表达均明显增加(P<0.05);与NEC组相比,NEC+BIF组新生小鼠建模3 d后体重明显增加,肠上皮损伤明显减轻,肠组织eNOS活性增强,O2^•含量明显减少,IL-6蛋白表达减少,IL-1β的mRNA及蛋白表达均减少(P<0.05),但NO含量差异无统计学意义(P>0.05)。结论双歧杆菌可减轻NEC新生小鼠肠组织损伤,其机制可能与肠组织eNOS活性增强有关。Objective To investigate the effect of Bifidobacterium on endothelial nitric oxide synthase and its potential protective mechanism in neonatal necrotizing enterocolitis using an induced mouse model.Methods Sixty 8-day newborn C57BL/6 mice were randomized into three groups(n=20):control group,necrotizing enterocolitis group(NEC)and Bifidobacterium intervention group(NEC+BIF).Samples were collected 3 days after NEC modeling.HE staining pathological score was used to evaluate the modeling effect;the level of endothelial nitric oxide synthase(eNOS)in the intestine was detected with Western blotting;the eNOS activity,nitric oxide(NO)and superoxide(O2^•)content were measured by Colorimetry.Real-time quantitative PCR was used to detect the mRNA level of interleukin-6(IL-6)and interleukin-1β(IL-1β);the protein expression of IL-6 and IL-1β was assessed by ELISA.Results No significant difference was observed in the protein levels of eNOS between the three groups.Compared with control group,in NEC group,the eNOS activity and NO content were significantly decreased;the O2^•content was substantially increased;both IL-6 and IL-1β were dramatically increased(P<0.05).As expected,compared with NEC group,mice in the NEC+BIF group showed a significant increase in weight with alleviation in intestinal epithelial damage.The eNOS activity was improved,the O2^• content was decreased,IL-6 was decreased in protein,IL-1β was decreased in both gene and protein level(P<0.05),but only a slight increase was observed in NO content(P>0.05).Conclusion Bifidobacterium can alleviate the severity of intestinal damage in NEC mouse model,the mechanism may be related to the enhancement of eNOS activity.

关 键 词：坏死性小肠结肠炎 双歧杆菌 内皮型一氧化氮合酶 

分 类 号：R574.5[医药卫生—消化系统]