高糖对大鼠心肌细胞NMDA受体表达、胞内钙和超氧自由基水平的影响.pdf

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1、医学研究杂志2014年10月第43卷第10期·论看·___J同宙糖对大鼠心肌细胞NMDA受体表达、胞内钙和超氧自由基水平的影响刘枭程秀丽郝维曹济民高雪摘要目的探讨高浓度葡萄糖对心肌细胞促离子型谷氨酸受体亚型NMDA受体(NMDAR)表达、胞内钙水平以及活性氧自由基产生的影响。方法分离新生(1—3天)大鼠心室肌细胞，心肌细胞与不同浓度葡萄糖(对照组葡萄糖浓度5mmoL／L，高糖组葡萄糖浓度33mmol／L)共孵育24—72h后，用Westernblot法检测心室肌细胞NMDAR1和NMDAR2B亚单位的表达水平，活细胞工作站

2、检测胞内ca“水平，流式细胞术检测细胞内活性氧自由基(ROS)含量。结果与对照组细胞相比，高浓度葡萄糖使心肌细胞的NMDAR2B的表达水平明显上调(P<0．05)，而NMDAR1的表达则降低。经高糖处理的心肌细胞在收缩时胞内钙水平高于对照细胞，高糖处理使心肌细胞的ROS水平升高，而用MK一801选择性阻断NMDAR可明显降低由高糖所致的胞内ROS增高(与高糖组对比，P<0．05)。结论高浓度葡萄糖可上调乳鼠心肌细胞NMDAR2B及下调NMDAR1的表达，并增加细胞内ca“水平及ROS的生成，而阻断NMDAR可抑制ROS的生

3、成。提示NMDAR可能参与糖尿病性心肌损害。关键词心肌细胞NMDA受体Ca“活性氧血糖[中图分类号]R3[文献标识码]AEfectsofHighGlucoseonNMDAReceptorExpression，IntracellularCa“andROSLevelsinCulturedNeonatalRatCardiomyocytes．LiuXiao，ChengXiuli，HaoWei，eta1．DepartmentofPhysiology&Pathophysiology，InstituteofBasicMedicalSci

4、encesChineseAcademyofMedicalSciences，SchoolofBasicMedicinePekingUnionMedicalCollege，Beqing100005，ChinaAbstract0bjectiveToinvestigatetheeffectsofhighglucoseonNMDAreceptor(NMDAR)expression，intracellularcalciumandreactiveoxygenspecies(ROS)levelsincardiomyocytes．Metho

5、dsHeartswereharvestedfromneonatalSDratsandventricularmyocyteswereisolatedandcuhured．WesternblotwasusedtoinvestigatetheexpressionsofNMDAR，confocalmicroscopywastodeter．minetheintracellularCa“level，flowcytometrytoexaminetheproductionofintracellularreactiveoxygenspeci

6、es(ROS)．ResultsComparedwiththecontrolcells(treatedwith5mmol／Lglucose)，incubationofcardiomyocyteswithhighglucose(33mmol／L)for72hin．creasedtheproteinexpressionofNMDAR2B，butdecreasedtheproteinlevelofNMDAR1(bothP<0．05)．HighglucosealsoelevatedtheintracellularCa“andROSl

7、evelsincardiomyocytes．SelectiveblockadeofNMDARbyMK一801(301~mol／L)suppressedtheelevationofROSinducedbyhighglucose．ConclusionHighglucoseupregulatesNMDAR2BexpressionbutdownregulatesNMDAR1expression，en—hancescalciummobilizationandROSproductioninculturedneonatalratcard

8、iomyocytes．BlockingNMDARsuppressedtheROSproductioninducedbyhighglucose．TheseresultssuggestedthatNMDARmaybeapharmacologica1targetfortreatingdiabeticcardi