·论著·肾小管间质纤维化与单核巨噬细胞的关系郭华△,邹万忠(北京大学基础医学院病理学系,北京100083)[关键词]肾小管/病理学;纤维化;单核细胞化学吸引蛋白质1;巨噬细胞集落刺激因子[摘要]目的:探讨肾小管间质纤维化与单核巨噬细胞(MC/MP)的关系。方法:结扎大鼠一侧肾静脉,连续饲养25d,制作大鼠肾小管间质纤维化模型。每5天杀检取材。HE、PAS、PASM、Masson等染色方法观察MC/MP的聚集和肾小管间质纤维化病变;免疫组化和双重免疫组化观察肾间质MC/MP的聚集与增生;原位杂交、免疫组化观察单核细胞化学吸引蛋白质1(MCP21)、巨噬细胞集落刺激因子(M2CSF)在肾内的表达分布。Westernblot检测病变肾脏MCP21的表达。对不同时间点肾小管变性、肾小管间质纤维化程度、肾间质MC/MP聚集的数量及其中的增生比例、MCP21和M2CSF的表达进行评估分析。结果:肾静脉结扎侧表现为肾小管变性萎缩、间质中MC/MP浸润、细胞外基质沉积等肾小管间质纤维化的典型病变。病变早期肾间质中有明显的MC/MP聚集,后期减少。表达增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)的单核细胞占一定比例,单核细胞的增生比例与聚集高度相关。原位杂交、免疫组化和Westernblot显示,病变肾组织中MCP21、M2CSFmRNA的表达主要位于变性的肾小管上皮细胞胞浆中,病变早期强,后期减弱。肾小管上皮细胞MCP21和M2CSF的表达与肾间质MC/MP的聚集高度相关。肾间质MC/MP的聚集与肾小管变性高度相关。结论:肾小管间质纤维化早期,间质中有明显的MC/MP聚集,来源于浸润和局部增生。肾小管上皮细胞通过表达MCP21、M2CSF在MC/MP浸润和增生中起了重要的调控作用。肾间质MC/MP的聚集与肾小管变性高度相关。MC/MP通过分泌多种细胞因子和促增殖因子促进肾小管间质纤维化,并进一步损伤肾小管。[中图分类号]R69216[文献标识码]A[文章编号]16712167X(2003)0520503205Relationshipbetweentubulointerstitialfibrosisandmononuclear2macrophageGUOHua△,ZOUWanzhong(DepartmentofPathology,PekingUniversitySchoolofBasicMedicalSciences,Beijing100083,China)KEYWORDSKidneytubules/pathol;Fibrosis;Monocytechemcattractantprotein21;Macrophagecolony2stimulatingfactorSUMMARYObjective:Tostudytherelationshipbetweenmonocyte/macrophage(MC/MP)accumu2lationandtubulointerstitialfibrosis.Methods:TherenaltubulointerstitialfibrosismodelinWistarratswasestablishedbyunilateralrenalveinligature.Theratswerefedfor25days.Thekidneyswereob2tainedevery5daysbykillingtherats.ThemorphologicalchangesoftubulointerstitialfibrosiswereobservedbylightmicroscopywithHE,PAS,PASMandMassonstaining.ImmunohistochemistryanddoubleimmunohistochemistrywereusedtoobserveMC/MPaccumulationandproliferation.InsituhybridizationandimmunochemistrywereusedtoobserveMCP21andM2CSFexpressioninexperimen2talrenaltissue.TheMCP21proteinexpressionwasinspectedbyWesternblot.Allthedatawereana2lyzedstatistically.Results:Thepathologicalchangesoftubulointerstitialfibrosisweretypical.ThereweremanyMC/MPsaccumulsatedintheinterstitialspaceattheearlystageandsomeofthemwerePCNApositive.AtthelatestagebothaccumulationandproliferationofMC/MPsweredecreased.TheportionofmonocyteproliferationwashighcorrelatedwiththeMC/MPaccumulation.Insituhy2bridizationshowedthepositivesignalsofMCP21andM2CSFweremainlylocatedinthecytoplasmofdegeneratedtubularepitheliumandtheywerestrongattheearlystage,weakatthelatestage.MCP21byimmunochemistryandWesternblotwereconsistentwithinsituhybridization.TheMC/MPac2cumulationwashighcorrelatedwiththeexpressionofMCP21andtubularepitheliumdegeneration.TheportionofmonocyteproliferationwashighcorrelatedwiththeexpressionofM2CSF.Conclusion:TherewasobviousaccumulationofMC/MPattheearlystageoftubulointerstitialfibrosis.Theaccu2mulationcamefrominfiltrationandproli...
