ecadherin和αsma基因的表观调控在肾间质纤维化中作用的分析word格式论文.docx

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ecadherin和αsma基因的表观调控在肾间质纤维化中作用的分析word格式论文.docx

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ecadherin和αsma基因的表观调控在肾间质纤维化中作用的分析word格式论文

Effect ofE-cadherinandalphaSMAgeneepigeneticregulation intheprocess of renalinterstitialfibrosis effectAbstract: Objective:PyrosequencingtechniquewasbeenusedfordetectionoftherelationshipofE-cadherinandalphaSMAgenepromoterregionmethylationstatusandgeneexpressionin the EMT process ofNRKcells,wecanmakein-depthunderstandingoftheroleofepigeneticintheEMTprocessofNRKcells,Itwillhelpustofurtherclarifythemechanismofrenalinterstitialfibrosis,andthusprovideanewdirectionforthepreventionandtreatmentofclinicalchronickidneydisease.Methods: Cultured normal NRK cells were divided into two groups:normalcontrolgroup、TGF-?1interventiongroup（10ng/ml）.Aftertwogroupsofcellswereinterventionsraised48h,themorphologyofcellswereobservedthroughinvertedphasecontrastmicroscope; theE-cadherinandα-SMAproteinexpressionlevelsofadherentcellsweresemi-quantitativedetectedbythecellularimmunocytochemicalmethod;thetypeIcollagenproteincontentofcellsculturesupernatantweredetectedbyELISA;extractedgenomicDNAfromcells,andtheE-cadherinandα-SMAmethylationlevelsofthetwogroupsofcellswererespectivelydetectedwiththepyrosequencing technology. TheCorrelationbetweenpromotermethylationandgeneexpressionwereanalysed. Results: 1) After 48h of cultured, compared with those ofnormalcontrol,thecellmorphology of TGF-?1intervention groupchangedsignificantly,manycellsshowedfibroblast-likemorphologychangedfromtheoriginaltypicalovalshapepavingstonelikeepithelialcell; 2)Immunocytochemistry demonstrated that the expression ofE-cadherinproteinweredecreasesignificantlyandtheexpressionof α-SMAproteinwereobviouslyincreasedinTGF-?1interventiongroup(P0.001).3)After48hofcultured,comparedwiththoseofnormalcontrol,thetypeIcollagencontentincreasedsignificantlyincellculturesupernatantofTGF-?1interventiongroup(P0.001).4)theE-cadherinandα-SMA gene promoter region methylation of two groups cells weredetectedbypyrosequencing,theresultshowedthatthepromoterregionmethylationofE-cadherinishigherinTGF-?1interventiongroupthancontrolgroup(P0.001),and