免疫性慢性非细菌性前列腺炎大鼠模型的形态学与分子生物学特性

114Vol.11No.420054NationalJournalofAndrologyApr.20051,1,2,3,3,3,3(1.,310031;2.,200030;3.,110032)::(CAP):,030d,51015mg/ml(FCA),45d,CAP:,(TNF2),21(IL21),IL22,(iNOS),,:15mg/mlFCA(11)1.0ml,0.5mlCAP:;;;:R2332;R697+233:A:100923591(2005)0420290206MorphologicalandMolecularBiologicalPeculiaritiesoftheExperimentalAutoimmuneProstatitisRatModelZHOUXiao2hui1,HANLei1,ZHOUZhi2heng2,LIUZhong2de3,YANGJi2xiang3,LUYan2wei3,YOUChun2lai31.ZhejiangUniversitySchoolofPharmaceuticalSciences,Hangzhou,Zhejiang310031,China;2.DepartmentofU2rology,LonghuaHospital,ShanghaiUniversityofTCM,Shanghai200030,China;3.DepartmentofUrology,theAffiliatedHospitalofLiaoningCollegeofTCM,Shenyang,Liaoning110032,ChinaCorrespondenceto:ZhouXiao2hui,E2mail:[e-mail]:Objective:Toobservethemorphologicalandmolecularbiologicalpeculiaritiesoftheexperimentalautoimmuneprostatitis(EAP)ratmodelmadebySCpurifiedprostateproteintwicewithimmuneadjuvant.Methods:Maleratswereintradermallyimmu2nizedwithasalineextractofmaleratprostateglands(RPG)inFreund’scompleteadjuvant(FCA)andPertussis2Diphtheria2Tetanusvaccine0.5mli.p.atthe0and30thday,andtheconcentrationsoftheextractwererespectively5mg/ml,10mg/mland15mg/ml.Atthe45thday,theratsweresacrificedandthemorphologicalandmolecularbiologicalchangesoftheprostatespecimenswereob2servedtodeterminetheeffectiveconcentrationofRPGforasuccessfulmodel.Results:TheexpressionofinflammationgenessuchasTNF2,IL21,IL22andiNOSobviouslyincreasedinthehigh2dosagemodelgroup;LM,EMandinsituhybridizationrevealedappear2antchronicinflammationresponse,butthiswasnotthecaseintheothertwodosagegroups.Conclusion:15mg/mlRPGmixedwithFCA(11)1.0mlSCwithPertussis2Diphtheria2Tetanusvaccine0.5mli.pwasaneffectivedosageforthesuccessfulmodelinourex2092:2004206215;:2004210212:(19712),,,,,,:,E2mail:[e-mail]©1994-2007ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(4):2902295Keywords:chronicabacterialprostatitis;animalmodel;morphology;molecularbiology(chronicabacterialpros2tatitis,CAP)1/42040,[1],,CAP,CAP,CAP11.11.1.13Wistar,,180220g,48,1.1.2(FCA):2:170,5mg/ml(),;:;(TNF2):,:200211215;(iNOS):;BcaBESTRNAPCRKitVer.1.1:(),:RR023CA;32(GAPDH):;DL22000DNAMarker:();X1742HaeDigestDNAMarker:();PCR:,:ForwardprimersBackwardprimersBasepairsofproductsIL2152GTCAACTATGTCCCGACCATT2352ATAGCAGCTTTCGACAGTGAG23416IL2252CTCTGAGCCCCAGCCTACCA2352AAAGCGGCTCCGAACACGAA23405GAPDH52GACATCAAGAAGGTGGTGAAG2352TGGAAATTGTGAGGGAGATGC233361.1.3(B62EMET2TLER);(JEM2100CX);(Bio2RadBenchmark);(DY892I);(JY922II);(DU2600);PCR(PEPE9600);(Bio2Rad1575);(1815TC,SAEL2LAB);(OlympusCHK2213);(HeraeusBiofuge28s)1.21.2.1240300gWistar10,,,,0.5%TritonX2100,,10000g,30min,,721,,,0.1mol/LpH7.2PBS51015mg/ml1.2.248(n=12)(n=12)(n=12)(n=10),:,0.5ml,FCA(11)1.0ml,5g/L,10g/L,15g/L0.9%0.51.0ml030d21.2.31.2.3.145d,,ELISATNF21.2.3.2,,1.2.41924©1994-2007ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(HE),1.2.4.31mm1mm1mm,2.5%1%,,Epon812,LKB,,1.2.4.4ELISATNF2490nm(A)AATNF21.2.4.5RT2PCR21(IL21)IL22mRNA50mg,1.2.4.6iNOS,iNOS1.3SPSS11.0;/;TNF2t22.1/,/,,11(xs)Table1.ThemorphologicalresultsoftheEAPrats(xs)GroupnMeanwetw.oftheprostate(mg)wetw./w.oftheprostate(%)PathologicalmanifestationControl12300.231.20.0900.009Flatsurface,integratedconstructionLow2dosagemodel12312.825.00.0950.006Low2gradehyperaemiaandedema,integratedconstructionMedium2dosagemodel12323.123.90.0990.0093Middle2gradehyperaemia,edemaandsynechiaeinconstructionofsometissueHigh2dosagemodel10358.832.7330.1090.01133Severehyperaemia,edemaandsynechiaeandconstructionofsometissueisnotclear,3:P0.05,33:P0.01;,:P0.01;,:P0.01Comparedwiththecontrolgroup,3:P0.05,33:P0.01;comparedwiththelow2dosagemodelgroup,:P0.01;comparedwiththemedium2dosagemodelgroup,:P0.012.2,;:,,,,,(,),(1);,2.3,;(,,,;),(2),:,,,2922005411©1994-2007ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(HE,400)Figure1.Prostatetissuesofthehigh2dosagemodelgroup(HE,400)2(EM,10000)Figure2.Prostatetissuesofthehigh2dosagemodelgroup(EM,10000)2.4ELISATNF2TNF2,22.5IL212.5.1IL21RT2PCR,IL21332TNF2(xs)Table2.TNF2contentinthebloodserumoftherats(xs)GroupnTNF2content(pg/ml)PControl1244.75.40.01High2dosagemodel1279.312.22.5.2IL21RT2PCR(/),IL21433IL21Table3.IL21contentintheprostatetissuesoftheratsGroupLaneDensityxControl2195185.53176High2dosagemodel4205205.552062.6IL22RT2PCR,,IL22,IL22543IL21RT2PCR1:DNA;25:IL21;69:GAPDHFigure3.IL21RT2PCRelectrophoresisstripsofthehigh2dos2agemodelgroupandthenormalcontrolgroupLane1:DNAmarker;Lanes25:IL21;Lanes69:GAPDH4IL21RT2PCRTable4.Semi2quantitativeratioofIL21intherats’prostatetissuesGroupLane(tested/ref.)Semi2quant.ratioMeansemi2quant.ratioControl2/60.92860.88743/70.8462High2dosagemodel4/80.98090.98335/90.98565IL22Table5.ExpressionintensityofIL22intherats’prostatetissueGroupLaneDensityxControl24344.5346High2dosagemodel4192195.051982.7iNOSiNOS,,3924©1994-2007ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.