植物雌激素鹰嘴豆芽素A（BCA）对肝纤维化去势小鼠模型的改善作用及机制.docx

DOI: 10. 12449/JCH240114肝纤维化及肝硬化植物雌激素鹰嘴豆芽素A(BCA)对肝纤维化去势小鼠模型的改善作用及机制谭超容a,李小飘a,冉俊艳a,熊英b,廖尚高a,张金娟b,何迅a贵州医科大学a.药学院，b.兽出医学院，c.慢性病诊疗转化工程研究中心，贵阳550025三f三情：Wt,2812878586(ORQD:0000-02-1867-069X);弓Wi,826609585(ORQD:0000-00002-9078-531X)摘要：目的探究植物雌激素鹰嘴豆芽素A(BCA)对CCL诱导的雌性双侧卵巢切除(去势)小鼠肝纤维化的影响及其机制。方法取50只去势雌性昆明小鼠，腹腔注射Cd建立肝纤维化模型，将建模小鼠按体质量随机分成模型组、阳性对照组、BCA低、中、高剂量组各10只，同时选取10只同批雌性小鼠切除双侧卵巢旁少量脂肪组织作为假手术组。假手术组和模型组灌胃等体积的05%枝甲娶F维索瞬液，阳飒照组®g二醇2mg/kg灌胃，BCA低中、高剂S组分另胺25、50、IoOmgkgBCA灌胃，1次/d,连续7周。给药结束后麻醉处死小鼠取材，测定肝|徽和子宫指数,HE及MaSSon染色观察肝组织病理改变,生化法检测AST、ALT活性，ELlSA法检测0干组织中IL6TNFC水平，WesternBIot法旋肝组织中I型胶原蛋白(CollagenI)、转化生长因子平1(TGF-1)、8平滑肌肌动蛋白QSMA)、雌激素受体(ER).p-NF-Bp65NF-Bp65蛋白的日寸表达量。计量资料两组间比较采用成组f检验，多组间比较采用单因素方差分析；进一步两两比较采用LSD-检验o非正态分布的计量资料多组间比较及进一步两两比较均采用Kruskal-WallisH检验。结果与假手术组相比，模型组肝指数升高，子宫指数降低，血清AST和ALT活性、肝组织中IL-6、TNFv水平及COiIagenI、TGF生CXSMA、pNF-Bp65NF*Bp65蛋白表达均升高(P值均<005),肝组织ER表达变化不明显(P>0.05).模型组肝组织明显纤维化病变,出现肝细胞水肿、脂肪样变、坏死并伴有炎细胞浸润，胶原纤维增生沉积、交错分布。与模型组比较，BCA各剂量组肝指数隔氐,血清ALT和AST活性、肝组织IL-6、TNFc水平及COllagenI、TGF-区、-SMA.P-NF-KBP65NF-Bp65的蛋白表达均降低(户值均0.05),子宫指数无明显变化(P>0.05),肝组织ER蛋白表达升高(P<0.05),肝组织的纤维化病变有不同程度改善»结论BCA可有效改善CCL诱导的雌性去势小鼠肝纤维化，其作用机制可能是通过上调ER抑制NF-KB信号通路，减轻炎症反应而实现的.关键词：肝纤维化；植物雌激素类；卵巢切除术；小鼠基金项目：贵州省高层次创新型人才"百层次人才"项目(黔科合平台人才20206011)RoleandmechanismofactionofphytoestrogenbiochaninAinimprovingliverfibrosisinOvariectomizedmiceTANChaOrOrd,LIXiaopiacf,RANJunyarf,XIONGYin,LIAOShanggacf,ZHANGJinjuarljc,HEXurf.(a.SchoolofPharmacy,b.SchoolofBasicMedicine,c.TransfomiationEngineeringResearchCenterofChronicDiseaseDiagnosisandTreatment,GuizhouMedicalUniversity,Guiyang550025,China)Correspondingauthor:HEXun,2812878586qq.m(ORCID:0000-0002-1867-069X),ZHANGJinjuan,826609585(ORCID:0000-00002-9078-531X)Abstract:OtyectiveToinvestigatetheeffectofphytoestrogenbiochaninA(BCA)onliverfibrosisinducedbyCCI4infemalemicewithbilateralphorectomy(Ovariectomized)anditsmechanism.MethodsAtotalof50OvariectomizedKunmingmicewereselectedandgivenintraperitonealinjectionofCCI4toestablishamodelofliverfibrosis,andthenacrdingtobodyweight,theywererandomlydividedintomodelgroup,positivecontrolgroup,andlow-,middle-,andhigh-doseBCAgroups,with10miceineachgroup.Inaddition,10femalemiceinthesameIftterweregivenresectionofasmallamountofadiposetissuenearbothovariestoestablishtheshamoperationgroup.Temiceinthepositivecontrolgroupweregivenestradiol2mg/kgbygavage,andthoseintelow-,middle-,andhighdoseBCAgroupsweregivenBCAbygavageatadoseof25,50,and100mg/kg,respectively,onceadayfor7ConSeeUtiVeweeks;themiceintheShamoPerationgroupandthemodelgroupweregivenanequalvolumeof05%sodiumcarboxymethylcellulosesolutionbygavage.Temicewereanesthetizedandsacrificedafteradministrationtocollectsamples.Liverindexanduterusindexweremeasured;HEstainingandMassonstainingWereusedtoobserveliverhistopathologicalchanges;thebiochemicalanalysiswasusedtomeasuretheactivityofaspartateaminotransferase(AST)andalanineaminotransferase(ALT);ELISAwasusedtomeasurethelevelsofinterleukin-6(IL-6)andtumorneosisfactor-(TNF-)inlivertissue,andWesternblotwasusedtomeasuretherelativeproteine×pressilevelsofcollagenI,transforminggrcwtfactor-beta1(TGF-1),alpha-smthmuscleactin(-SMA),estrogenreceptorbeta(ER),andp-NF-Bp65NF-Bp65inlivertissue.7eWestwasusedforcomparisonfcontinuousdatabetweentwogroups;aone-wayanalysisofvariouswasusedforcomparisonbetweenmultiplegroups,andtheleastsignificantdifferenceAtestwasusedforKirthercomparisonbetweentwogroupsTheKnJSkalWallisHtestwasusedforcomparisofnon-nomallydistributedCOntinUOUSdatabetweenmultiplegroupsandfurthercomparisonbetweentwogroupsResultsComparedwithteShamopefatedgroup,themodelgrouphadasignificantineaseinliverindexandasignificantreductioninuterusindex,aswellassignificantincreasesintheactivitiesofserumASTandALT,televelsofl-6andTNF-inlivertissue,andtheproteinexpressionlevelsofcollagenI,TGF,-SMA,andp-NF-Bp65NF-Bp65inlivertissue(allP<0.05),withnosignificantchangeintheexpressionofERinlivertissue(P>0.05),andthemodelgroupshowedsignificantfibrosislesionsintheliver,suchashepatocyteedema,steatosis,andnecrosiswithinflammatorycellinfitrationandhyperplasia,deposition,andstaggereddistributionofcollagenfibers.ComparedVththemodelgroup,thelew-,middle-,andhigKdoseBCAgroupshadsignificantreductionsinliverindex,theactivitiesofsermASTandALT,thelevelsofIL-6andTNF-,andtheproteinexpressionlevelsofcollagenI,TGF-1,-SMA,andp-NF-Bp65NF-Bp65inlivertissue(all?<0.05),whnosignificantchangeinuterineindex(P>0.05),aswellasasignificantincreaseintheproteinexpressionlevelofERinlivertissue(9<0.05)andvaryingdegreesofimprovementinliverfibrosislesions.CdusionBCAcaneffectivelyimproveCCL,-inducedliverfibrosisinQvariectomizedfemalemice,possiblybyUpregulatingERtoinhibittheNF-Bsignalingpathwayandthenalleviatinginflammatoryresponse.Keywords:HepaticFibrosis;Phytoestrogens;Ovariectomy;MiceResearchfunding:GuizhouProvincialHigh-LevelInnovativeTalentswHundredLevelTalentsaProject(GuizhouKehePlatformTalents20206011)肝纤维化是各种慢性肝病发展为肝硬化及肝癌的必经阶段，在早期阶段