本课题组以往临床和实验研究显示附子及附子为主的经验方改善心功能和心室重构，前期对其水溶性强心单体去甲乌药碱（Higenamine）的体外研究发现其通过β2/Gi/PI3K/AKT途径抑制新生大鼠、成年小鼠心肌细胞氧化应激引起的凋亡，同时发现该药物同样导致成年小鼠心肌细胞肥大。在此基础上，我们拟采用体外灌流小鼠心脏、单个心肌细胞荧光共振能量转移（FRET）技术证明Higenamine通过β2兴奋细胞膜局部cAMP/PKA活性发挥强心作用；采用缺血再灌注和心梗后心衰小鼠模型及体外RNA干扰技术证明其通过β2/Gi/PI3K/AKT通路在体内及体外抗心肌细胞凋亡，保护心功能；采用小鼠主动脉缩窄（TAC）模型及体外RNA干扰、药物抑制的方法证明其诱导的心肌细胞肥大是有良性代偿意义的生理性肥大而非病理性肥大，其通路也是β2/Gi/PI3K/AKT。完善附子的现代分子生物学研究，以期开发心衰治疗新药。

In our previous study, we found Chinese herb "Aconite root" also named "fuzi" and Chinese medical prescriptions that characterized by using "fuzi" as monarch, can improve the cardiac function and remodeling in human and rat. Higenamine , as the main inotropic functional compound of "fuzi", was demonstrated recently in vitro by our group to attenuate apoptosis through beta2/Gi/PI3K/AKT pathway. Interestingly, we also found Higenamine induced hypertrophy in primary cultured adult mouse ventricular myocytes. Based on preliminary data, we hypothesize that: [1]. Higenamine plays inotropic role in heart via exciting beta2/Gs and membrane- specific cAMP/PKA pathway, which leads to cardiac inotropic. [2]. By activating beta2/Gi/PI3k/AKT pathway, Higenamine attenuates cardiac apoptosis in vivo and in vitro. [3]. Also through this signal pathway, Higenamine triggers physiological cardial hypertrophy, which may be complementary benefit in later heart failure. To test our hypothesis, first, ex-vivo Langendorff heart perfusion system and fluorescence resonance energy ransfer(FRET) will be utilized to measure beta2 AR/Gs, membrane specific AC/cAMP/PKA pathway and inotropic function of Higenamine. Secondly, ischemia/reperfusion mouse model and myocardial infarction model in vivo, siRNA technology to knock down target gene in vitro will be performed to confirm the anti-apoptotic function of Higenamine. Finally, mouse transverse aortic constriction (TAC) model in vivo, pharmacological inhibitor and RNAi knock down in vitro will be done to support that: Higenamine cause physiological instead of pathological cardiac hypertrophy through beta2/Gi/PI3K/AKT pathway. This findings will provide strong evidence of inotropic, anti-apoptotic and physiological hypertrophy inducible function of Higenamine in later stage of heart failure, and Higenamine may have potential to develop new candidate for therapy of heart failure.