植物乳杆菌活菌体和热灭活菌体对大肠杆菌黏附IPEC-J2细胞的影响
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摘要
【目的】探讨植物乳杆菌活菌体和热灭活菌体对猪肠上皮细胞(intestinal epithelial cells,IPEC-J2细胞)的黏附特性,以及对大肠杆菌黏附IPEC-J2细胞的抑制作用.【方法】分别用植物乳杆菌活菌体和热灭活菌体处理IPEC-J2细胞,结合显微镜观察和黏附计数方法评估其黏附特性;评价其在竞争、排阻和置换3种方式下对大肠杆菌黏附IPEC-J2细胞的影响.【结果】植物乳杆菌活菌体和热灭活菌体均能黏附于IPEC-J2细胞,其中活菌体对IPEC-J2细胞的黏附指数较热灭活菌体高,但差异不显著(P>0.05);活菌体对大肠杆菌黏附IPEC-J2细胞的竞争抑制和置换抑制率显著高于热灭活菌体(P<0.05);两种生物学状态的植物乳杆菌对大肠杆菌黏附IPEC-J2细胞的排阻抑制率差异不显著(P>0.05).【结论】植物乳杆菌活菌体和热灭活菌体均能黏附于IPEC-J2细胞,并可通过竞争、排阻和置换方式抑制大肠杆菌对IPEC-J2细胞的黏附,且其活菌体对大肠杆菌黏附IPEC-J2细胞的竞争和置换抑制效果优于热灭活菌体.
【Objective】To investigate the adhesion of live and heat-inactivated Lactobacillus plantarum(L.plantarum)on IPEC-J2 cells and the effect of live and heat-inactivated on the inhibition of adhesion of Escherichia coli to IPEC-J2 cells.【Method】The IPEC-J2 cells were treated with live and heat-inactivated L.plantarum and then the adhesion of them was assessed using light microscope observation and plate count methods.The ability of live and heat-inactivated L.plantarumto inhibit the E.coli adhesion on IPEC-J2 cells by competitive,exclusion and displacement was also investigated.【Result】Live and heat-in-activated L.plantarumcould adhere to the IPEC-J2 cells.Although the adhesion index of live bacteria was higher than the adhesion index of heat-inactivated bacteria on IPEC-J2 cells,but the difference was not significant on them(P>0.05).The live bacteria showed a significantly better inhibition ratio than heat-inactivated bacteria about E.coli adhesion on IPEC-J2 cells(P<0.05).The inhibition ratio of E.coli adhesion on IPEC-J2 cells was not significant difference on live and heat-inactivated bacteria(P>0.05).【Conclusion】Live and heat-inactivated L.plantarumcould adhere to IPEC-J2 cells and reduce the adhesion of E.coli to IPEC-J2 cells by competitive,exclusion and displacement.Furthermore,the effect of competitive and displacement of live L.plantaruminhibited E.coli adhesion on IPEC-J2 cells were superior to heat-inactivated L.plantarum.
【Objective】To investigate the adhesion of live and heat-inactivated Lactobacillus plantarum(L.plantarum)on IPEC-J2 cells and the effect of live and heat-inactivated on the inhibition of adhesion of Escherichia coli to IPEC-J2 cells.【Method】The IPEC-J2 cells were treated with live and heat-inactivated L.plantarum and then the adhesion of them was assessed using light microscope observation and plate count methods.The ability of live and heat-inactivated L.plantarumto inhibit the E.coli adhesion on IPEC-J2 cells by competitive,exclusion and displacement was also investigated.【Result】Live and heat-in-activated L.plantarumcould adhere to the IPEC-J2 cells.Although the adhesion index of live bacteria was higher than the adhesion index of heat-inactivated bacteria on IPEC-J2 cells,but the difference was not significant on them(P>0.05).The live bacteria showed a significantly better inhibition ratio than heat-inactivated bacteria about E.coli adhesion on IPEC-J2 cells(P<0.05).The inhibition ratio of E.coli adhesion on IPEC-J2 cells was not significant difference on live and heat-inactivated bacteria(P>0.05).【Conclusion】Live and heat-inactivated L.plantarumcould adhere to IPEC-J2 cells and reduce the adhesion of E.coli to IPEC-J2 cells by competitive,exclusion and displacement.Furthermore,the effect of competitive and displacement of live L.plantaruminhibited E.coli adhesion on IPEC-J2 cells were superior to heat-inactivated L.plantarum.
引文
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[22]许荣誉,庄建良,张海云.12株乳杆菌益生特性的研究[J].医生研究生学报,2008,21(12):1246-1249.
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[24]邢咏梅,贾继辉,王红艳,等.两种生物状态肠道益生菌的粘附和粘附拮抗效应的对比研究[J].中国微生态学杂志,2004,16(2):69-72.
[25]孙建芝,贾继辉,曲伟,等.多种乳酸杆菌粘附活性的研究[J].医学检验与临床,2006,17(5):38-40.
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[28]NSTAD S N,SALARIAN A A,GHAHRAMANI M H,et al.Live and heat-inactivated Lactobacilli from feces inhibit Salmonella typhi and Escherichia coli adherence to Caco-2cells[J].Folia Microbiologica,2009,54(2):15-160.
[29]丁琴凤,马丽,冯希平.嗜酸乳杆菌及其灭活菌粘附及拮抗牙周致病菌特性研究[J].现代口腔医学杂志,2012,26(6):378-382.
[30]谯仕彦,侯成立,曾祥芳.乳酸菌对猪肠道屏障功能的调节作用及其机制[J].动物营养学报,2014,26(10):3052-3063.
[2]MENNIGEN R,BRUEWER M.Effect of probiotics on intestinal barrier function[J].Annals of the New York Academy of Sciences,2009,1165:183-189.
[3]NANDA KUMAR N S,BALAMURUGAN R,JAYAKANTHAN K,et al.Probiotic administration alters the gut flora and attenuates colitis in mice administered dextran sodium sulfate[J].Journal of Gastroenterology and Hepatology,2008,23(12):1834-1839.
[4]OHLAND C L,MACNAUGHTON W K.Probiotic bacteria and intestinal epithelial barrier function[J].American Journal of Physiology:Gastrointestinal and Liver Physiology,2010,298(6):G807-G819.
[5]NIKOAKELAINEN S,SALMINEN S,BYLUND G,et al.Characterization of the properties of human-and dairy-derived probiotics for prevention of infectious diseases in fish[J].Applied and Environmental Microbiology,2001,67(6):2430-2435.
[6]PETERSON L W,ARTIS D.Intestinal epithelial cells:regulators of barrier function and immune homeostasis[J].Nature Reviews Immunology,2014,14(3):141-153.
[7]FUJIWARA S,HASHIBA H,HIROTA T,et al.Inhibition of the binding of enterotoxigenic Escherichia coli Pb176to human intestinal epithelial cell line HCT-8by an extracellular protein fraction containing BIF of Bifidobacterium longum SBT2928:suggestive evidence of blocking of the binding receptor gangliotetraosylceramide on the cell surface[J].International Journal Food Microbiology,2001,67(1/2):97-106.
[8]白洁,李卫芬,黄琴,等.几株益生乳酸菌对Caco-2细胞的黏附及其对致病菌黏附的影响[J].动物营养学报,2012,24(10):1992-1998.
[9]SCHIERACK P,NORDHOFF M,POLLMANN M,et al.Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine[J].Histochemistry and Cell Biology,2006,125(3):293-305.
[10]ZHNAG W,ZHU Y,YANG J,et al.A selected Lactobacillus rhamnosus strain promotes EGFR-Independent Akt activation in an enterotoxigenic Escherichia coli K88-infected IPEC-J2cell model[J].畜牧与饲料科学,2015,10(Z1):27-42.
[11]陈德龙,许光勇,任晓明,等.乳杆菌竞争性抑制两种病原菌粘附上皮细胞的研究[J].中国农业科学,2012,45(14):2991-2998.
[12]侯成立.罗伊氏乳杆菌全基因组序列分析及其调节仔猪肠黏膜免疫功能的研究[D].北京:中国农业大学,2015.
[13]俞涵丽,方华,蔡旋,等.嗜酸乳杆菌活菌体和热灭活菌体对IPEC-J2细胞的黏附和黏附拮抗效应[J].动物营养学报,2014,26(6):1601-1607.
[14]任清霞,张文卿,孙建芝.双歧杆菌对肠道上皮细胞的粘附和粘附拮抗效应的研究[J].医学检验与临床,2009,20(2):21-23.
[15]LIU Y,FATHEREE N Y,MANGAALAT N,et al.Human-derived probiotic Lactobacillus reuteristrains differentially reduce intestinal inflammation[J].American Journal of Physiology:Gastrointestinal and Liver Physiology,2010,299(5):G1087-G1096.
[16]王大会,齐雪峰,赵献军,等.猪源乳酸杆菌对永生化猪小肠上皮细胞的黏附性能[J].中国兽医学报,2012,32(7):980-983.
[17]CAMPANA R,FEDERICI S,CIANDRINI E,et al.Antagonistic activity of Lactobacillus acidophilusATCC 4356on the growth and adhesion/invasion characteristics of human Campylobacter jejuni[J].Current Microbiology,2012,64(4):371-378.
[18]倪学琴,JOSHUA GONG,曾东,等.猪源乳酸杆菌对鼠伤寒沙门氏菌DT104在猪肠道上皮细胞IPEC-J2上粘附的影响[J].微生物学通报,2008,35(7):1072-1077.
[19]HORI K,MATSUMOTO S.Bacterial adhesion:from mechanism to control[J].Biochemical Engineering Journal,2010,48(3):424-434.
[20]张永刚,DAVIN R.猪小肠上皮细胞作用的新发现[J].中国畜牧杂志,2015(8):57-60.
[21]陈臣,郭本恒,王荫榆,等.植物乳杆菌ST-Ⅲ粘附肠上皮样细胞系Caco-2性质的研究[J].乳业科学与技术,2008(2):51-55.
[22]许荣誉,庄建良,张海云.12株乳杆菌益生特性的研究[J].医生研究生学报,2008,21(12):1246-1249.
[23]李孝辉,皮雄娥,许尧兴.热灭活嗜酸乳杆菌培养物在饲料工业中的应用[J].饲料与畜牧,2009(12):14-18.
[24]邢咏梅,贾继辉,王红艳,等.两种生物状态肠道益生菌的粘附和粘附拮抗效应的对比研究[J].中国微生态学杂志,2004,16(2):69-72.
[25]孙建芝,贾继辉,曲伟,等.多种乳酸杆菌粘附活性的研究[J].医学检验与临床,2006,17(5):38-40.
[26]PARSOT C,SANSONETTI P.Invasion and the pathogenesis of Shigellainfections[J].Current Topics in Microbiology and Immunology,1996,209:25-42.
[27]CROXEN M A,FINLAY B B.Molecular mechanisms of Escherichia coli pathogenicity[J].Nature Reviews Microbiology,2010,8(1):26-38.
[28]NSTAD S N,SALARIAN A A,GHAHRAMANI M H,et al.Live and heat-inactivated Lactobacilli from feces inhibit Salmonella typhi and Escherichia coli adherence to Caco-2cells[J].Folia Microbiologica,2009,54(2):15-160.
[29]丁琴凤,马丽,冯希平.嗜酸乳杆菌及其灭活菌粘附及拮抗牙周致病菌特性研究[J].现代口腔医学杂志,2012,26(6):378-382.
[30]谯仕彦,侯成立,曾祥芳.乳酸菌对猪肠道屏障功能的调节作用及其机制[J].动物营养学报,2014,26(10):3052-3063.