用共培养体系观察缺氧肾小管上皮细胞对成纤维细胞的调节作用

THEMODULATIONOFRENALTUBULAREPITHELIALCELLSTREATEDWITHHYPOXIAONRENALINTERSTITIALFIBROBLASTSINCOCULTURE

为了解肾小管细胞与间质成纤维细胞的相互关系在肾小管间质病变中的作用，利用不直接接触细胞共培养方法在体外观察了缺氧的小鼠近端肾小管上皮细胞对肾小管周围成纤维细胞增殖、分泌细胞外基质及表达细胞间粘附分子－１（ＩＣＡＭ－１）的影响。结果显示，缺氧后的肾小管上皮细胞可促进：（１）肾小管周围成纤维细胞的增殖。（２）成纤维细胞分泌纤维连结蛋白（ＦＮ）及层粘连蛋白（ＬＮ）明显增加（吸光度Ａ），ＦＮ：０．７０８±０．０２８比０．６０５±０．０２４；ＬＮ：０．７１８±０．０３２比０．５９１±０．０２７，Ｐ＜０．０１。（３）成纤维细胞膜上ＩＣＡＭ－１的表达增强，４４．８６±２．２２比４１．２２±２．０４，Ｐ＜０．０１。结果提示缺氧后的肾小管上皮细胞对其周围成纤维细胞具有明显的调节作用。

Toclarifycel-celinteractioninthepathogenesisofrenaltubulointerstitialnephritis(TIN),theefectofrenaltubularepithelialcels(TEC)onrenalinterstitialperitubularfibroblasts(PTF)wasexaminedincelcoculturesystemwithoutdirectcontact.TECandPTFwerepreparedfromthekidneyofBALB/Cmice.Firstly,TECwereplatedintointercupchambersof24-welplatesandin-cubatedat37℃inahumidified5%CO2atmospherefor72hours.Thenthecelswereculturedinanin-cubatorwhichwasfulof95%N2for24hours.Secondly,thoseintercupswithhypoxia-treatedTECwerefloatedonthewelsof24-welplatescontainingPTFwhichhadbeenincubatedfor24hours.TheTECandthePTFwerecoculturedforanother48hours.PTFwerealsococulturedwithnormalTECascontrols.TheparametersofthecoculturedPTFweremeasuredasfolows.(1)Celproliferationexam-inedbyMTTincorporationmethodandthecelnumbersweredetectedwithtotalacidphosphataseac-tivity.(2)Fibronectin(FN),laminin(LN)andcolagenⅣinthesupernatantsofcoculturesystemweremeasuredbyELISAmethod.(3)ThedistributionofICAM-1onthecelmembraneofPTFwasdeterminedbylaserconfocalscanningmicroscopy.Results:(1)ThenumberofPTFcoculturedwithhypoxiatreatedTECwasgreaterthanthatofcontrols(P<0.01).(2)ThelevelofFNandLNinthesupernatantsofthePTFcoculturedwithhypoxia-treatedTECwashigherthanthatofthecontrols(P<0.05).(3)TheexpressionofICAM-1onPTFcoculturedwithhypoxia-treatedTECwashigherthanthatofcontrols(P<0.01).InconclusiontheinteractionofPTFandTECmayplayaroleinthepatho-genesisofTIN.