奶山羊瘤胃上皮细胞紧密连接蛋白mRNA表达量的影响 .pdfVIP

体外法研究pH与脂多糖或组胺的交互作用对奶山羊瘤胃上皮

细胞紧密连接蛋白mRNA表达量的影响

孙燕勇;高民;徐明;宋利文;李洋;李超;陈利青;胡红莲

【摘要】Theobjectiveofthisstudywastoinvestigatetheinteraction

effectsofpHandlipopolysaccharide(LPS)orhistamine(HIS)onmRNA

expressionlevelsoftightjunctionproteinsofrumenepitheliumofdairy

goatsinvitro.EighthealthySaanendairygoatswithsimilarbodyweight

andmilkyieldwereusedasthedonorsofrumenepithelium.A3×3two-

factorexperimentdesignwasadopted.Afterslaughtering,rumen

epitheliumwascollectedandinsertedinthemiddleofhalfchamberina

Ussingchamber.Bufferat5mLwasaddedtotheserosalside,and

differenttreatmentsofculturemediumsat5mLwereaddedtothe

mucosalside,eachtreat-menthad3replicates.Experiment1,factor1was

pH,andwas7.4,5.5,5.2,respectively;factor2wasLPSconcentration,andwas

0,30and60kEU/mL,respectively.Inexperiment2,factor1waspH,andwas

7.4,5.5,5.2,respectively;factor2wasHISconcentration,andwas0,0.5and

10.0ng/mL,respectively.Therumenepitheliumwascollectedafter

culturedfor80min,andtheexpressionlevelsofClaudin-1,Claudin-4,

Claudin-7,Occludingandzonulaoccludens-1(ZO-1)mRNAsweremeasured.

Theresultsshowedasfollows:1)theinteractionbetweenpHandLPShad

significanteffectsonexpressionlevelsofClaudin-1,Claudin-7andZO-1

mRNAs(P0.05).ComparedwithpH7.4×0kEU/mLLPSgroup,the

decreaseofpHorthesupple-mentationofLPSsignificantlydecreased

expressionlevelsofClaudin-1andClaudin-7mRNAs(P0.05),andtended

toriseexpressionlevelofZO-1mRNA,whileitwasthehighestinpH

5.2×60kEU/mLLPSgroup.2)TheinteractionbetweenpHandHIShad

significanteffectsonexpressionlevelsofClaudin-1,Claudin-7andZO-1

mRNAs(P0.05).pH5.5×0.5ng/mLHISgrouphadthelowestexpression

levelofClaudin-1mRNA,whichwasnotsignificantlydifferentfromthatin

pH5.2×0.5ng/mLHISgroup(P0.0