从调控Akt/p53、Akt/Hif-1α信号通路及串话探讨参蛤散抗心力衰竭的作用机制

Heart failure, with high prevalence and poor prognosis, is of unsatisfied curative effect in modern medicine. Under the guidance of the Traditional Chinese Medicine（TCM） theory of treating different diseases with same method, we found Shen’ge Powder favorable functions on heart failure. Previous studies demonstrated that Shen’ge Powder could inhibit cardiomyocyte apoptosis, promote angiogenesis, and activate Akt, Hif-1α/VEGF, down-regulate p53. But the precise mechanism involved is not yet clearly elucidated. This study is to detect the effect of Shen’ge Powder on heart function and angiogenesis after the establishment of pressure overload heart failure in rat, with CD31 marking vessels in myocardium. Separate the nuclear and cytoplasmic protein and test Akt, p53, Hif-1α and VEGF. Establish cardiomyocyte injury model by Doxorubicin to up-regulated p53. With the intervention of LY294002 and Pifithrin-α to down-regulate Akt, p53, explore the effect of Shen’ge Powder on the regulation of Akt/p53, Akt/Hif-1α pathway, the nuclear translocation and crosstalk of p53、Hif-1α. This study will provide experimental data support for Shen’ge Powder on heart failure and TCM theory of treating different diseases with same method.

心力衰竭患病率高、预后差，现代医学疗效不佳。在“异病同治”中医理论指导下，参蛤散有良好的抗心衰作用。前期研究表明参蛤散可抑制心肌凋亡、促进血管生长，能激活Akt和Hif-1α/VEGF，并下调p53，但其具体调控机制尚不明确。本研究拟通过建立压力超负荷心衰大鼠模型，CD31标记心肌中血管，观察参蛤散对心肌凋亡、心肌血管生长的作用，分离细胞核和细胞浆蛋白，检测Akt、p53、Hif-1α、VEGF等指标。同时，体外以阿霉素建立心肌细胞损伤模型，诱导p53上调，使用LY294002、Pifithrin-α等工具药下调Akt、p53，进一步研究参蛤散对Akt/p53、Akt/Hif-1α通路的调控机制和对p53、Hif-1α的转核、串话影响。为参蛤散治疗心力衰竭和中医“异病同治”理论提供实验数据支持。

心力衰竭是各种心脏疾病的终末阶段，发病率高，治疗手段有限，预后不佳。在“异病同治”理论指导下，我们前期临床和基础研究发现参蛤散具有很好的抗心力衰竭作用，对Akt、Hif-1α、p53、VEGF等具有调控作用，但具体机制尚不明确。本研究建立压力超负荷心衰大鼠模型，发现参蛤散可明显改善心功能，提高心肌中血管数量和VEGF受体，促进Akt磷酸化，上调Hif-1α，抑制p53等凋亡因子。细胞实验中，我们亦发现参蛤散可以明显抑制阿霉素诱导H9C2心肌细胞的凋亡，使用LY294002、Pifithrin-α等工具药后，发现该作用是通过激活PI3K/Akt，从而上调Hif-1α和抑制p53，并且可能通过抑制p53而进一步上调Hif-1α。另外参蛤散可以明显促进Hif-1α转核和抑制p53转核。研究结果说明参蛤散可通过抑制凋亡和促进血管生长两方面发挥抗心力衰竭的作用，机制与Akt/p53、Akt/Hif-1α通路有关，从而证明了参蛤散抗心力衰竭的有效性，亦为祖国医学“异病同治”理论提供了一定的实验支持。