苍术素对大鼠心脏正性肌力的作用及其机制  

Positive inotropic effect of atractylodin in normal rats and its underlying mechanism

作　　者：高丽 张文慧 王羽维 朱晓佳 肖钰洁 高倩雯 李伟[1] 陈龙[1,3] GAO Li;ZHANG Wen-hui;WANG Yu-wei;ZHU Xiao-jia;XIAO Yu-jie;GAO Qian-wen;LI Wei;CHEN Long(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023;Taizhou Fourth People’s Hospital,Taizhou 225300;Institute of Chinese Medicine of Taizhou China Medical City,Taizhou 225300,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]泰州市第四人民医院,江苏泰州225300 [3]泰州中国医药城中医药研究院,江苏泰州225300

出　　处：《中国应用生理学杂志》2020年第5期408-413,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金项目(81573304);泰州中国医药城第四批“113人才计划”(2016024);江苏省研究生科研创新计划(KYCX19_1301,KYCX19_1328)。

摘　　要：目的:研究中药苍术的有效成分苍术素(Atractylodin)的正性肌力作用及其机理。方法:随机选取6只雄性SD大鼠进行在体压力-容积环(P-V loop)实验,加药前为Control组,经腹腔注射苍术素(3 mg/kg)后为苍术素组(自身对照),分析6只雄性大鼠加药后对大鼠左心室心输出量、容积及动脉压的作用;大鼠离体心脏灌流实验中,依次灌流给药:第一部分为Control→0.1→1→10μmol/L苍术素浓度梯度灌流,第二部分为Control→200 nmol/L H89(PKA抑制剂)→200 nmol/L H89+10μmol/L苍术素,第三部分为Control→500 nmol/L KN-93(CaMKII抑制剂)→500 nmol/L KN-93+10μmol/L苍术素,第四部分为Control→10 nmol/L Calyculin A(PP1,PP2A抑制剂)→10 nmol/L Calyculin A+10μmol/L苍术素,加药前的正常空白组为Control组,分析每部分各6只雄性大鼠的组间左心室发展压的变化;在大鼠心肌细胞钙释放实验中,分组、给药的方法和浓度同离体心脏实验,分析来源于6个雄性大鼠6个左心室心肌细胞组间钙释放幅值的变化。结果:①P-V loop实验表明:与Control组相比,苍术素组(3 mg/kg)腹腔注射30 min后,显著增加大鼠心输出量、搏出功及心率(P<0.05),降低动脉舒张压(P<0.05),而对收缩压无明显影响;②离体心脏实验表明:与Control组相比,苍术素组(0.1,1,10μmol/L)灌流10 min后,能显著增加大鼠离体心脏左心室发展压(LVDP)(P<0.05),其作用能被H89组(200 nmol/L)所阻断;③心肌细胞钙释放实验表明:与Control组相比,苍术素组(10μmol/L)灌流10 min后,基于肌浆网钙泵(SERCA2a)显著增加大鼠心肌细胞钙释放幅值(P<0.05),其作用能被H89组(200 nmol/L)所阻断。结论:苍术素具有正性肌力作用,其血流动力学特点表现为降低在体大鼠动脉舒张压而不增加收缩压,苍术素的正性肌力作用机制是通过PKA-SERCA2a通路发挥的。Objective:To explore the positive inotropic effect of atractylodin which is major active component of Rhzoma Atractylodis Lanceae and its underlying mechanism.Methods:For in vivo study,six male SD rats were randomly selected for the heart pressure-volume loop(P-V loop)experiment.The effects of atractylodin(3 mg/kg,intraperitoneal injection)on hemodynamic parameters such as LVDP(left ventricular developed pressure),SW(stroke work),HR(heart rate),CO(cardiac output),SBP(systolic blood pressure)and DBP(diastolic blood pressure)were analyzed.For in vitro study,left ventricular developed pressure(LVDP)from the Langendroff-perfused isolated rat heart was analyzed before as the control and after atractylodin perfusion.For in vitro study,the effects of atractylodin and atractylodin with H89(PKA inhibitor)or KN-93(CaMKII inhibitor or Calyculin A(PP1,PP2A inhibitor)on LVDP were analyzed.The experiments were separated into four parts with six isolated hearts for each as follows:(1)Control→0.1→1→10μmol/L atractylodin;(2)Control→200 nmol/L H89(PKA inhibitor)→200 nmol/L H89+10μmol/L atractylodin;(3)Control→500 nmol/L KN-93(CaMKII inhibitor)→500 nmol/L KN-93+10μmol/L atractylodin;(4)Control→10 nmol/L Calyculin A(PP1,PP2A inhibitor)→10 nmol/L Calyculin A+10μmol/L atractylodin.For the study of rat left ventricular myocyte Ca 2+transient induced by field stimulation,the experiment design was the same as in vitro study.The six cells from the different rats were used for each part experiment.Results:①Atractylodin(3 mg/kg)significantly increased the heart rate,cardiac output and stroke work(P<0.05)and decreased the diastolic blood pressure(P<0.05).②Atractylodin(0.1,1,10μmol/L)significantly increased the LVDP in a concentration dependent manner(P<0.05).The positive inotropic effect of atractylodin could be blocked by PKA inhibitor H89.③Atractylodin(10μmol/L)significantly increased the amplitude of SR Ca 2+transient amplitude mediated by facilitating sarcoplasmic reticulum SERCA2a.The enhanced amplitude of

关 键 词：苍术素 Ghrelin受体 正性肌力 PKA 钙释放 肌浆网钙泵 大鼠 

分 类 号：R969.4[医药卫生—药理学]