1-磷酸鞘氨醇受体途径诱导的心肌肥大及蛋白激酶C的调节作用  

Sphingosine-1-Phosphate-Protein Kinase C Signaling Regulates the Cardiomyocyte Hypertrophic Responses Induced by Sphingosine-1-Phosphate

作　　者：李跃艳[1] 熊升林[2] 

机构地区：[1]南华大学附属第一医院心血管内科,湖南省衡阳市421001 [2]南华大学心血管疾病研究所,湖南省衡阳市421001

出　　处：《中国动脉硬化杂志》2011年第7期573-577,共5页Chinese Journal of Arteriosclerosis

摘　　要：目的研究1-磷酸鞘氨醇受体途径在1-磷酸鞘氨醇(S1P)诱导乳鼠心肌细胞肥大反应中的作用。方法原代培养乳鼠心肌细胞,测定心肌细胞体积和[3H]-亮氨酸掺入量作为心肌细胞肥大的指标。实验分对照组(不加任何干预因素)、S1P组(S1P直接作用于心肌细胞)、VPC23019组(S1P1和S1P3受体抑制剂进行干预30 min后,加入S1P刺激心肌细胞)、JTE组(S1P2受体抑制剂进行干预30 min后,加入S1P刺激心肌细胞)及Staurospo-rine组(蛋白激酶C抑制剂进行干预30 min后,加入S1P刺激心肌细胞),[3H]-亮氨酸掺入法测定心肌细胞蛋白合成速率;用qPCR和Western blot法检测心肌细胞β-肌球蛋白重链的mRNA和蛋白表达水平。结果 10、100和1000 nmol/L的S1P均能增加乳鼠心肌细胞的细胞体积和[3H]-亮氨酸掺入量,尤以1000 nmol/L S1P最为明显,因而选择1000 nmol/L S1P作用心肌细胞48 h为成功的心肌细胞肥大模型。1000 nmol/L S1P处理心肌细胞48 h,可使[3H]-亮氨酸掺入量明显增加,该作用可被S1P2受体抑制剂或蛋白激酶C抑制剂明显抑制。与对照组相比,S1P组心肌细胞β-肌球蛋白重链的mRNA和蛋白表达水平显著增加。与1000 nmol/L S1P组相比,S1P2受体抑制剂组和特异性蛋白激酶C抑制剂组β-肌球蛋白重链的mRNA和蛋白表达水平显著下降。结论 S1P介导的心肌肥大主要依赖S1P2途径介导。S1P2介导心肌肥大反应的机制可能部分通过激活蛋白激酶C途径而实现。Aim To investigate the regulatory role of sphingosine-1-phosphate receptors （S1PR） in cardiomyocyte hypertrophicresponse induced by S1P. Methods Cellvolumes and protein compound rate of neonatal heart cells were used as indexs of hypertrophic cardiomyocytes models induced by SIP. Neonatal heart ceils were randomly divided into 5 groups： normal control; hyperrophic group： ceils were stimulated by S1P （1000 nmol/L） ; pretreatment group： cells were treated with VPC23019 （10 μmol/L） 30 min before adding S1P; pretreatment group： cells were treated with JTE（ 10 μmol/L） 30 rain before adding S1P and pretreatment group： cells were treated with Staurosporine（ 10 nmol/L） 30 min before adding S1P. Protein compound rate of neonatal rat cardiomyocyte ceils was assayed by [ 3 H ] -leucine incorporation. The expression of β-myosin heavy chain （β-MHC） was detected by qPCR and Western Blot . Results 10, 100 and 1000 nmol/L S1P increased neonatal rat cardiac cell volume and [ 3H ] - leucine incorporation, especially 1000 nmol/ L SIP. Cells treated with SIP ht at（100 nmol/L for 48 h were chosen as hypertrophic cardiocyte model. S1P （ 1000 nmol/ L） treatment of myocytes 48 h, can increase significantly [ 3H ] - leucine incorporation, the effect was significantly inhibited by S1P2 receptor inhibitor（JTE） or protein kinase C （PKC） inhibitor（Staurosporine）. Compared with the control group, hypertrophy of myocardial cells of β-MHC mRNA and protein levels were significantly increased. Compared with SIP （ 1000 nmol/L） treatment group, β-MHC expression levels of S1P2 receptor-specific inhibitor and the PKC inhibitor group was significantly decreased. Conclusion The activation of S1P2 played an important role in cardiomyocytes hypertrophy induced by S1P. S1P2-PKC signaling regulates the cardiomyocyte hypertrophic responses induced by SIP.

关 键 词：蛋白激酶C 心肌细胞肥大 1-磷酸鞘氨醇 

分 类 号：R363[医药卫生—病理学]