人参皂苷Rb1调节钙、钠通道影响受损心肌细胞的研究  

Research on Effect of Ginsenoside Rb1 on Damaged Myocardial Cells by Regulating Calcium and Sodium Channels

作　　者：王冰冰 孔宏亮[2] WANG Bingbing;KONG Hongliang(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Liaoning Provincial People's Hospital Liaoning,Shenyang 110067,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁省人民医院,辽宁沈阳110067

出　　处：《实用中医内科杂志》2024年第10期125-128,I0013-I0015,共7页Journal of Practical Traditional Chinese Internal Medicine

基　　金：沈阳市科技创新专项(A19-112-4-057)。

摘　　要：目的 探讨人参皂苷Rb1(Gs-Rb1)减轻阿霉素(Dox)诱导的心肌细胞损伤,明确Gs-Rb1改善Dox心肌细胞损伤效应与其调节钠、钙通道有关,AMP依赖性蛋白激酶(AMPK)和/或钙调蛋白激酶Ⅱ(CaMKII)通路介导该作用。方法 提取大鼠左心室心肌细胞进行培养,将细胞随机分为7组:对照组(PBS)、Dox组、Gs-Rb1组、AraA(阿糖腺苷,AMPK抑制剂)组、AraA+Gs-Rb1组、KN93(CAMKII抑制剂)组及KN93+Gs-Rb1组,分别进行给药。ELISA法检测N末端B型利钠肽前体、AMPK和CaMKⅡ活性;DCFH-DA荧光探针试验检测线粒体内活性氧(mitROS)水平,流式细胞仪检测细胞内活性氧(ROS)水平;Western blot检测总LTCC(t-LTCC)、L型电压门控钙通道(LTCC)、兰尼定受体(RYR2)、肌浆网钙ATP酶(SERCA2a)和钙钠交换体(NCX)蛋白表达。结果 Gs-Rb1显著降低心肌细胞NT-proBNP水平,该作用可被AraA显著抑制;Gs-Rb1升高心肌细胞AMPK活性被AraA完全抑制,不被KN93影响;Gs-Rb1降低心肌细胞CaMKII活性;Gs-Rb1降低细胞内ROS和mitROS水平;Gs-Rb1升高SERCA2a蛋白表达,降低NCX表达量;各组tLTCC差异无明显统计学意义(均P>0.05)。结论 Gs-Rb1降低Dox诱导心肌细胞NT-proBNP水平,通过调节LTCC再分布、增强RYR2表达、增加SERCA2a和抑制NCX等减轻心肌细胞损伤,AMPK或/和CaMKII通路介导该效应;Gs-Rb1显著抑制Dox诱导的心肌细胞氧化应激,此效应可被AMPK通路和/或CaMKII通路介导。Objective To investigate whether ginsenoside Rb1(Gs-Rb1)can alleviate doxorubicin(Dox)induced myocardial cell injury,and to determine whether the improvement effect of Gs-Rb1 on Dox myocardial cell injury is related to its regulation of sodium and calcium channels,and whether the AMP dependent protein kinase(AMPK)and/or calmodulin kinase II(CaMKII)pathways mediate this effect.Method Rat left ventricular myocardial cells were extracted and cultured.The cells were randomly divided into 7 groups:control group(PBS),Dox group,Gs-Rb1 group,AraA group,AraA+Gs-Rb1 group,KN93 group,KN93+Gs-Rb1 group.These groups were administered separately.ELISA method was used to detect the activities of N-terminal B-type natriuretic peptide precursor,AMPK and CaMKII.DCFH-DA fluorescence probe assay was used to detect mitochondrial reactive oxygen species(mitROS)levels,and flow cytometry was used to detect intracellular reactive oxygen species(ROS)levels.Western blot was used to detect the expressions of total LTCC(t-LTCC),L-type voltage gated calcium channel(LTCC),ranidine receptor(RYR2),sarcoplasmic reticulum calcium ATPase(SERCA2a)and calcium sodium exchanger(NCX)proteins.Results The results showed that Gs-Rb1 significantly reduced the level of NT-proBNP in myocardial cells,which was significantly inhibited by AraA.Gs-Rb1 increased the AMPK activity of myocardial cells,which was completely inhibited by AraA and not affected by KN93.Gs-Rb1 reduced CaMKII activity in myocardial cells.Gs-Rb1 reduced intracellular ROS and mitROS levels.Gs-Rb1 increased SERCA2a protein expression and reduced NCX expression.There was no statistically significant difference in tLTCC among the groups(all P>0.05).Conclusion Gs-Rb1 reduces Dox induced NT-proBNP levels in myocardial cells,alleviating myocardial cell damage by regulating LTCC redistribution,enhancing RYR2 expression,increasing SERCA2a and inhibiting NCX.AMPK or/and CaMKII pathways mediating this effect.Gs-Rb1 significantly inhibits Dox induced oxidative stress in cardiomyocytes,which can be medi

关 键 词：心肌细胞 人参皂苷RB1 离子通道 氧化应激 

分 类 号：R541.6[医药卫生—心血管疾病]