甲状旁腺激素对HAEC细胞EndMT诱导作用及机制探讨.pdf

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1、垒生旦i箍墨鲞茎旦基目JBloodPurif，September，2014，Vo1．9，No．13·647··基础研究·甲状旁腺激素对HAEC细胞EndMT诱导作用及机制探讨李宁李亚斐【摘要】目的探讨甲状旁腺激素(parathyroidhormone，PTH)是否诱导血管内皮细胞上皮细胞一间充质转化(endothelial—mesenchymaltransition，EndMT)及相关机制。方法血管内皮细胞HAEC培养至对数期，分别加入不同剂量的全长重组PTH(10mol／L、10mol／L、10omol／L、10mol／L、10-smo1／L)作用48h；再

2、在10mol／LPTH作用下，选择12、24、36、60h4个时间点；分别收集总蛋白。Westernblot分析Ve—Cadherin、CD31、ct—SMA、TGF—B1和ILK的表达改变。结果Westernblot结果表明在PTH作用下，血管内皮细胞标志分子We—Cadherin和CD31表达降低；纤维细胞标志分子a—SMA表达则显著升高，且呈时间和剂量依赖性。Westernblot分析发现TGF—B1和ILK表达同时升高。结论PTH减弱内皮细胞特征、增强纤维化特征，诱导HAEC细胞发生EndMT，TGF-卜ILK通路是其调控的可能通路之一。【关键词】甲状

3、旁腺素；内皮细胞．间充质转化；尿毒症；整合素连接激酶中图分类号：R318．16文献标识码：Adoi：10．3969／j．issn．1671—4091．2014．09．0101[1leefectandmechanismofparathyroidhormoneinducedendothelia1．mesenchymaltransitionofvascularell-dotheliMHAECcellsLINing,LIl一feiInstituteofPreventiveMedicine,ThethirdMilitaryMedicalUni-versi~Chongqi

4、ng40003DepartmentofNephrology,刀FirstAfiliatedHospital,刀thirdMilitaryMedl"-calUniversi~Cbongqing400038．ChinaCorrespondingauthOF：LIl厂aEmail：liyafei2OO8@hotmail,corn[Abstract]ObjectiveToinvestigatewhethertheincreasedparathyroidhomaone(PTH)asauremictox．ininducestheendothelial—mesenchyma

5、ltransitionfEnd／vlT)ofvascularendothelialcellsfHACEcells)anditsrelatedmechanisms．MethodsThevascularendothelialHACEcellswereculturedtologarithmicphase．andPTHattheconcentrationof10-12，10一l1，10．10，l0．9，and10．8mol／Lwasaddedintothemediumfor48h．Additionally,HACEcellswereculturedinthemediu

6、mcontaining10．8mol／LPTHfor12，24，36．and60h．CellswerethenharvestedformorphologicalexaminationandforVECadherin，CD31，a．SMA．TGF一131andILKexpressiondeterminationsbywesternblot．ResuitsMorphologicalexaminationshowedthattheHACEcellstendedtohavefibrosischangesafterthePTHinduction．Wstemblotdis

7、playedthatPTHreducedtheex．pressionofvascularendothelialcellmarkersCD31andVECadherinandincreasedtheexpressionoffibercellmarkersa-SMAintimEanddosEdependentmanners．AtthesametimetheexpressionsofTGF．131andILKiB．creased．ConclusionPTHinducedtheHAECcellstohavelessendothelialcellcharacterist

8、icsandmorefi．brosis