ERK/P38MAPK信号转导通路在人脐动脉血管平滑肌细胞表型转化中的作用  

Effects of ERK/P38MAPK pathway on the phenotypic modulation of human umbilical arterial smooth muscle cell

作　　者：纵振坤[1] 李中林[1] 范月超[1] 刘勇[1] 

机构地区：[1]徐州医学院附属医院神经外科十病区,221002

出　　处：《中华脑血管病杂志（电子版）》2009年第2期4-8,共5页Chinese Journal of Cerebrovascular Diseases(Electronic Edition)

摘　　要：目的探讨血小板源生长因子-BB(PDGF-BB)及米诺环素是否可通过调节ERK/P38MAPK信号通路,从而影响人脐动脉血管平滑肌细胞(HASMC)的表型转化。方法建立HASMC体外培养模型,分为无血清的DMEM、PDGF-BB(20ng/ml)、PDGF-BB(20ng/ml)+PD98059(30μmol/L)+SB203580(20μmol/L)、PDGF-BB(20ng/ml)+米诺环素(15μmol/L)、PDGF-BB(20ng/ml)+米诺环素(30μmol/L)五组,Western Blot法检测ERK1/2、P-ERK1/2、P38、P-P38蛋白表达。结果体外培养的HASMC在PDGF-BB(20ng/ml)+米诺环素(15μmol/L)培养液孵育24h后,P-P38蛋白表达较PDGF-BB组明显下降(P<0.01);在PDGF-BB(20ng/ml)+米诺环素(30μmol/L)组,P-ERK1/2和P-P38蛋白表达均较PDGF-BB组明显下降(P<0.01),表明米诺环素显著抑制ERK/P38MAPK信号通路。结论(1)PDGF-BB诱导HASMC的去分化与ERK/P38MAPK信号通路有关,如抑制ERK/P38MAPK信号通路的活性,则HASMC保持分化表型;(2)米诺环素对PDGF-BB诱导HASMC去分化的抑制作用是通过抑制ERK/P38MAPK信号通路的活性,下调PDGF-BB诱导的ERK1/2和P38磷酸化水平而实现的,与其对HASMC的细胞毒性无关。Objective To investigate the role and the molecular mechanisms of the ERK and p38 MAPK pathways on the phenotypic modulation of the cultured vascular smooth muscle cells. Methods To establish a steady cell culture model in vitro for vascular smooth muscle cells （VSMC） of human umbilical artery. The HASMC was incubated with DMEM without serum, PDGF-BB （20 ng/ml）, PDGF-BB （20 ng/ml） ＋ PD98059 （30 μmol/L） ＋SB203580 （20μmol/L）, PDGF-BB （20 ng/ml） ＋ minocycline （15 μmol/L ）, PDGF-BB （20 ng/ml ） ＋ minocycline （30μmol/L ）. The expression of ERK1/2, P- ERK1/2, P38 and P- P38 protein of HASMC was detected by the way of Western Blot. Results The relative expression of P- P38 protein of HASMCs incubated with PDGF-BB （20 ng/ml） ＋ minocycline （ 1 5μmol/L） for 24h, was lower than that of the PDGF-BB （20 ng/ml） group （P〈0.01）. The relative expression of P-ERK1/2, P- P38 protein of HASMCs incubated with PDGF-BB （20 ng/ml） ＋ minocycline （ 15μmol/L）,PDGF-BB （20 ng/ml） ＋ minocycline （30μmol/L） for 24h,was respectively lower than that of the PDGF-BB （20ng/ml） group, revealing that minocycline down-regulated P-ERK1/2, P- P38 protein of HASMC significantly （P〈0.01）. Conclusions （1） PDGF-BB activated ERK/P38MAPK pathways to induced de-differentiation of the HASMC. When the ERK/P38MAPK pathways were simultaneously blocked by their specific inhibitofs, PDGF-BB in turn initiated to maintain the differentiated SMC phenotype. （2） This result suggests that the inhibitory effect of minocycline on HASMC de-differentiation induced by PDGF-BB was not caused by the cytotoxicity caused by minocycline, but because it depressed ERK / P38MAPK pathways and down-regulated the expression of P-ERK1/2, P- P38 protein.

关 键 词：血管平滑肌细胞 表型转化 米诺环素 ERK1/2 P38MAPK 

分 类 号：R543[医药卫生—心血管疾病]