摘要
目的观察慢性低氧诱导的系膜细胞增殖，细胞外基质ｍＲＮＡ表达及蛋白激酶Ｃ（ＰＫＣ）在低氧介导系膜细胞增殖中的作用。方法采用体外大鼠系膜细胞培养，Ｎｏｒｔｈｅｒｎｂｌｏｔ分析及放射核素测定法。结果与正常氧（１８％Ｏ２）比较，低氧（３％Ｏ２）培养系膜细胞３Ｈ－胸腺嘧啶掺入率显著增高（３．３±０．３×１０３ｍｉｎ－１／瓶对１．７±０．２×１０３ｍｉｎ－１／瓶，Ｐ＜０．０１）。细胞计数在培养２８小时无显著差异，但培养７２小时，低氧比正常氧培养增加３３％（Ｐ＜０．０１）。低氧培养系膜细胞表达Ⅳ型前胶原（ＰＣｏｌⅣ）、纤维连接蛋白（Ｆｎ）、层粘连蛋白（Ｌａ）ｍＲＮＡ显著增高。测定低氧培养系膜细胞ＰＫＣ活力呈双峰型增高，加入ＰＫＣ抑制剂Ｈ７可抑制低氧诱导的系膜细胞增殖。结论慢性低氧可诱导系膜细胞增殖和细胞外基质产生增多。
bjectiveToexaminetheefectofchronichypoxiaontheproliferationofculturedratglomerularmesangialcelsandexpressionofextracelularmatrix.MethodsCelularproliferationwasassessedusing[3H]-thymidineincorporationandcelnumber,expressionoftypeⅣprocolagen,fibronectinandlamininwasanalysedbyNorth-ernblot.ActivityofproteinkinaseC(PKC)wasdeterminedbyradionucleartechnique.ResultsSubconfluentquiescentcelsexposedtohypoxiafor28hoursexhibitedasignificantincreasein[3H]-thymidineincorporationfol-lowedbyasignificantincreaseincelnumberat72hoursincomparisonwithrespectivenormoxiccontrols.Themesangialcelsexposedtohypoxiashowedasignificantincreaseinexpressionofextracelularmatrixcomparedwithnormoxiccondition.HypoxiainducedbiphasicactivationofPKC,furthermore,chronichypoxic-inducedstim-ulationof[3H]-thymidineincorportionwascompletelypreventedbyaninhibitorofPKC.ConclusionChronichypoxiainducestheproliferationofculturedglomerularmesangialcelsandincreaseinexpressionofextracelularmatrixmediatedbytheactivationofPKC.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
1996年第6期352-355,共4页
Chinese Journal of Nephrology
