阿如拉-7味散对感染致病性E.coli O_1大鼠肠黏膜屏障的影响
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摘要
作者旨在研究阿如拉-7味散对感染致病性E.coli O1大鼠肠黏膜机械屏障、免疫屏障和化学屏障的影响。选用60只6～8周龄SD大鼠进行试验,分为空白对照组、未治疗组、环丙沙星组和阿如拉-7味散高(1.08g·kg~(-1))、中(0.54g·kg~(-1))、低(0.27g·kg~(-1))剂量组,共6组,每组10只。试验结束时,计算其保护率,并采集小肠各段进行HE染色,测定小肠黏膜形态指标;同时采集血清和回肠组织,ELISA试剂盒法检测血清中的D-乳酸和二胺氧化酶(DAO)含量以及回肠黏膜中的分泌型免疫球蛋白(sIgA)和肠三叶因子(ITF)含量。结果表明:(1)阿如拉-7味散中剂量对感染致病性E.coli O_1大鼠的保护率最好,为50%;且对小肠黏膜形态有一定的改善和修复作用,其效果与环丙沙星相当;(2)环丙沙星组与空白对照组相比D-乳酸含量无显著差异(P>0.05);阿如拉-7味散高、中剂量组DAO含量与空白对照组相比无显著差异(P>0.05);(3)除阿如拉-7味散低剂量组外,其余各组与未治疗组相比回肠黏膜sIgA含量显著提高(P<0.05);未治疗组大鼠回肠黏膜ITF含量与其余各组相比显著降低(P<0.05)。结果提示,阿如拉-7味散对感染致病性E.coli O_1大鼠引起的肠黏膜损伤有一定的修复作用,其中中剂量(0.54g·kg~(-1))效果最佳,与抗生素环丙沙星的修复效果相当。
This study was conducted to investigate the effect of Arula-7 powder on mechanical barrier,immune barrier and chemical barrier in rats infected with Escherichia coli O1.A total of 60 SD rats,6-8weeks old,were equally divided into the control group,untreated group and ciprofloxacin group,Arula-7 powder high(1.08 g·kg~(-1)),medium(0.54 g·kg~(-1))and low(0.27g·kg~(-1))dosage groups(n=10).The rate of protection was calculated.We collected the small intestine and the intestinal morphology indexes were measured by HE staining.Meanwhile,the serum and ileal tissue were also collected and the levels of D-lactic acid,DAO,sIgA and in-testinal trefoil factor were detected using ELISA kit.The results showed that:(1)Arula-7 powder with medium dose had the highest protection rate of 50%.Furthermore,the medium dose of Arula-7powder had positive effects on the improvement or recovery of the morphology of intestinal mucosa in rats infected with E.coli O_1 and its effect was similar with that of ciprofloxacin.(2)There was no significant difference between the control group and the ciprofloxacin group in the level of D-lactic acid(P>0.05).The level of DAO had no significant difference between the control group and the Arula-7powder medium/high dosage groups(P>0.05).(3)Except for Arula-7powder low dosage group,the significant level of sIgA was observed in ileal mucosa of other four groups compared with the untreated group(P<0.05).The level of ITF in ileal mucosa of untreated group was significantly lower than other medicine treatment groups(P<0.05).In conclusion,Arula-7 powder has the best effect for repairing the intestinal mucosa injury in rats infected with E.coli O_1 at medium dose(0.54g·kg~(-1)),and its effect is similar as that of the antibiotic ciprofloxacin.
This study was conducted to investigate the effect of Arula-7 powder on mechanical barrier,immune barrier and chemical barrier in rats infected with Escherichia coli O1.A total of 60 SD rats,6-8weeks old,were equally divided into the control group,untreated group and ciprofloxacin group,Arula-7 powder high(1.08 g·kg~(-1)),medium(0.54 g·kg~(-1))and low(0.27g·kg~(-1))dosage groups(n=10).The rate of protection was calculated.We collected the small intestine and the intestinal morphology indexes were measured by HE staining.Meanwhile,the serum and ileal tissue were also collected and the levels of D-lactic acid,DAO,sIgA and in-testinal trefoil factor were detected using ELISA kit.The results showed that:(1)Arula-7 powder with medium dose had the highest protection rate of 50%.Furthermore,the medium dose of Arula-7powder had positive effects on the improvement or recovery of the morphology of intestinal mucosa in rats infected with E.coli O_1 and its effect was similar with that of ciprofloxacin.(2)There was no significant difference between the control group and the ciprofloxacin group in the level of D-lactic acid(P>0.05).The level of DAO had no significant difference between the control group and the Arula-7powder medium/high dosage groups(P>0.05).(3)Except for Arula-7powder low dosage group,the significant level of sIgA was observed in ileal mucosa of other four groups compared with the untreated group(P<0.05).The level of ITF in ileal mucosa of untreated group was significantly lower than other medicine treatment groups(P<0.05).In conclusion,Arula-7 powder has the best effect for repairing the intestinal mucosa injury in rats infected with E.coli O_1 at medium dose(0.54g·kg~(-1)),and its effect is similar as that of the antibiotic ciprofloxacin.
引文
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[15]贺琴,王自蕊,游金明,等.酵母壁多糖对断奶仔猪生长性能和小肠黏膜形态结构的影响[J].动物营养学报,2016,28(11):3536-3541.HE Q,WANG Z R,YOU J M,et al.Effects of yeast cell wall polysaccharides on growth performance and small intestinal mucosa morphology of weaned piglets[J].Chinese Journal of Animal Nutrition,2016,28(11):3536-3541.(in Chinese)
[16]邓军,李云锋,杨倩.枯草芽孢杆菌和猪源乳酸杆菌混合饲喂对仔猪肠绒毛发育的影响[J].畜牧兽医学报,2013,44(2):295-301.DENG J,LI Y F,YANG Q.Effects of co-administration of Bacillus subtilis and porcine Lactobacillus salivarius on intestinal villus of piglets[J].Acta Veterinaria et Zootechnica Sinica,2013,44(2):295-301.(in Chinese)
[17]钟武装,宋卫兵,杨键,等.炎症性肠病患者黏附分子-l、D-乳酸、二胺氧化酶的变化及临床意义[J].现代消化及介入诊疗,2013,18(5):273-276.ZHONG W Z,SONG W B,YANG J,et al.Clinical significance of sICAM-1,plasma diamine oxidase and D-lactate in inflammatory bowel disease patients[J].Modern Digestion&Intervention,2013,18(5):273-276.(in Chinese)
[18]呙于明,刘丹,张炳坤.家禽肠道屏障功能及其营养调控[J].动物营养学报,2014,26(10):3091-3100.GUO Y M,LIU D,ZHANG B K.Intestinal barrier of poultry:function and modulation[J].Chinese Journal of Animal Nutrition,2014,26(10):3091-3100.(in Chinese)
[19]孙勇,王良喜.三叶因子家族研究进展[J].肠外与肠内营养,2015,22(1):49-52.SUN Y,WANG L X.The research progress of trefoil factor family[J].Parenteral&Enteral Nutrition,2015,22(1):49-52.(in Chinese)
[20]谢天宇,胡红莲,高民.肠黏膜免疫屏障及其保护措施[J].动物营养学报,2014,26(5):1157-1163.XIE T Y,HU H L,GAO M.Gut mucosal immune barrier and the protective measures[J].Chinese Journal of Animal Nutrition,2014,26(5):1157-1163.(in Chinese)
[21]XIAO P,LING H,LAN G,et al.Trefoil factors:gastrointestinal-specific proteins associated with gastric cancer[J].Clin Chim Acta,2015,450:127-134.
[22]孙必强,伍参荣,周英,等.不同剂型七味白术散对肠道菌群失调小鼠小肠黏膜超微结构和sIgA的影响[J].中国微生态学杂志,2016,28(2):125-128,137.SUN B Q,WU C R,ZHOU Y,et al.Effects of different formulations of Qiwei Baizhu powder on the ultrastructure of small intestinal mucosa and sIgA in mice with intestinal dysbacteriosis[J].Chinese Journal of Microecology,2016,28(2):125-128,137.(in Chinese)
[23]苏敏.肠安Ⅱ号方对三硝基苯磺酸(TNBS)诱导炎症后肠易激综合征(PI-IBS)大鼠肠粘膜免疫屏障的作用机制研究[D].北京:中国中医科学院,2014.SU M.Changanerhao decoction for the treatment of trinitrobenzene sulfonic acid(TNBS)induced post-inflammation irritable bowel syndrome(PI-IBS)involving intestinal mucosal immune barrier in a rat model[D].Beijing:China Academy of Chinese Medical Sciences,2014.(in Chinese)
[2]MIRHOSEINI A,AMANI J,NAZARIAN S.Review on pathogenicity mechanism of enterotoxigenic Escherichia coli and vaccines against it[J].Microb Pathog,2018,117:162-169.
[3]HU J,TORRES A G.Enteropathogenic Escherichia coli:foe or innocent bystander?[J].Clin Microbiol Infect,2015,21(8):729-734.
[4]LIU L L,WU W,ZHANG J Y,et al.Progress of research on the toxicology of antibiotic pollution in aquatic organisms[J].Acta Ecol Sin,2018,38(1):36-41.
[5]IANIRO G,TILG H,GASBARRINI A.Antibiotics as deep modulators of gut microbiota:between good and evil[J].Gut,2016,65(11):1906-1915.
[6]杨斯琴,敖日格乐,王纯洁,等.蒙药对牛源致病性大肠杆菌的体外抑菌效果研究[J].中国农业大学学报,2015,20(1):124-128.YANG S Q,AO-RI-GE-LE,WANG C J,et al.Antibacterial effects of Mongolian medicines on bovine pathogenic Escherichia coli in vitro[J].Journal of China Agricultural University,2015,20(1):124-128.(in Chinese)
[7]杨斯琴,敖日格乐,王纯洁,等.3种蒙药复方对致病性Escherichia coli O1的抑菌机理及对小鼠的保护率[J].微生物学通报,2016,43(8):1766-1773.YANG S Q,AO-RI-GE-LE,WANG C J,et al.Inhibition of pathogenic Escherichia coli O1and protection of infected mice by three Mongolian medicinal compounds[J].Microbiology China,2016,43(8):1766-1773.(in Chinese)
[8]姚媛媛.致犊牛腹泻病肠道大肠杆菌的分离鉴定及蒙药复方对其抗菌效果研究[D].呼和浩特:内蒙古农业大学,2014.YAO Y Y.A study on the isolation and identification of E.coli from calf diarrhea and antimicrobial effect of Mongolian medicine on they[D].Hohhot:Inner Mongolia Agricultural University,2014.(in Chinese)
[9]战永波.治疗奶牛乳房炎蒙药复方的筛选及其抗炎免疫机理的研究[D].呼和浩特:内蒙古农业大学,2010.ZHAN Y B.Screening of compound Mongolian medicine treating bovine mastitis and research of its mechanism on anti-inflammation and immunoenhancement[D].Hohhot:Inner Mongolia Agricultural University,2010.(in Chinese)
[10]李盼.中草药复合制剂对仔猪大肠杆菌性腹泻治疗效果研究[D].长春:吉林农业大学,2015.LI P.Study of therapeutic effect on Chinese herbal compound preparation for Escherichia coli-induced diarrhea of piglet[D].Changchun:Jilin Agricultural University,2015.(in Chinese)
[11]王东旭.白头翁散在控制仔猪大肠杆菌性腹泻中的应用效果研究[D].洛阳:河南科技大学,2017.WANG D X.Study on the application effect of Pulsatilla pulvis in the prevention and control of diarrhea induced by Escherichia coli[D].Luoyang:Henan University of Science and Technology,2017.(in Chinese)
[12]陈鹏举.穿黄散对产ESBLs大肠杆菌的抑菌作用及临床效果研究[D].长春:吉林大学,2015.CHEN P J.Study on the Bacteriostatic action and clinical effect of Chuanhuang powder against ESBLsproducing E.coli[D].Changchun:Jilin University,2015.(in Chinese)
[13]OLIVARES-VILLAGOMEZ D,VAN KAER L.Intestinal intraepithelial lymphocytes:sentinels of the mucosal barrier[J].Trends Immunol,2018,39(4):264-275.
[14]XUE Z G,YU J C,ZHAO M L,et al.Effects of synbiotics on intestinal mucosal barrier in rat model[J].Clin Nutr Exp,2017,13:12-21.
[15]贺琴,王自蕊,游金明,等.酵母壁多糖对断奶仔猪生长性能和小肠黏膜形态结构的影响[J].动物营养学报,2016,28(11):3536-3541.HE Q,WANG Z R,YOU J M,et al.Effects of yeast cell wall polysaccharides on growth performance and small intestinal mucosa morphology of weaned piglets[J].Chinese Journal of Animal Nutrition,2016,28(11):3536-3541.(in Chinese)
[16]邓军,李云锋,杨倩.枯草芽孢杆菌和猪源乳酸杆菌混合饲喂对仔猪肠绒毛发育的影响[J].畜牧兽医学报,2013,44(2):295-301.DENG J,LI Y F,YANG Q.Effects of co-administration of Bacillus subtilis and porcine Lactobacillus salivarius on intestinal villus of piglets[J].Acta Veterinaria et Zootechnica Sinica,2013,44(2):295-301.(in Chinese)
[17]钟武装,宋卫兵,杨键,等.炎症性肠病患者黏附分子-l、D-乳酸、二胺氧化酶的变化及临床意义[J].现代消化及介入诊疗,2013,18(5):273-276.ZHONG W Z,SONG W B,YANG J,et al.Clinical significance of sICAM-1,plasma diamine oxidase and D-lactate in inflammatory bowel disease patients[J].Modern Digestion&Intervention,2013,18(5):273-276.(in Chinese)
[18]呙于明,刘丹,张炳坤.家禽肠道屏障功能及其营养调控[J].动物营养学报,2014,26(10):3091-3100.GUO Y M,LIU D,ZHANG B K.Intestinal barrier of poultry:function and modulation[J].Chinese Journal of Animal Nutrition,2014,26(10):3091-3100.(in Chinese)
[19]孙勇,王良喜.三叶因子家族研究进展[J].肠外与肠内营养,2015,22(1):49-52.SUN Y,WANG L X.The research progress of trefoil factor family[J].Parenteral&Enteral Nutrition,2015,22(1):49-52.(in Chinese)
[20]谢天宇,胡红莲,高民.肠黏膜免疫屏障及其保护措施[J].动物营养学报,2014,26(5):1157-1163.XIE T Y,HU H L,GAO M.Gut mucosal immune barrier and the protective measures[J].Chinese Journal of Animal Nutrition,2014,26(5):1157-1163.(in Chinese)
[21]XIAO P,LING H,LAN G,et al.Trefoil factors:gastrointestinal-specific proteins associated with gastric cancer[J].Clin Chim Acta,2015,450:127-134.
[22]孙必强,伍参荣,周英,等.不同剂型七味白术散对肠道菌群失调小鼠小肠黏膜超微结构和sIgA的影响[J].中国微生态学杂志,2016,28(2):125-128,137.SUN B Q,WU C R,ZHOU Y,et al.Effects of different formulations of Qiwei Baizhu powder on the ultrastructure of small intestinal mucosa and sIgA in mice with intestinal dysbacteriosis[J].Chinese Journal of Microecology,2016,28(2):125-128,137.(in Chinese)
[23]苏敏.肠安Ⅱ号方对三硝基苯磺酸(TNBS)诱导炎症后肠易激综合征(PI-IBS)大鼠肠粘膜免疫屏障的作用机制研究[D].北京:中国中医科学院,2014.SU M.Changanerhao decoction for the treatment of trinitrobenzene sulfonic acid(TNBS)induced post-inflammation irritable bowel syndrome(PI-IBS)involving intestinal mucosal immune barrier in a rat model[D].Beijing:China Academy of Chinese Medical Sciences,2014.(in Chinese)
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