基于AMPK/ULK1信号通路研究温阳活血化痰法通过调控自噬防治蒽环类心脏毒性的作用机制

Effect of anthracycline well in leukemia, breast cancer and other malignant tumors were present, especially in the treatment of breast cancer plays an irreplaceable role, but its serious cardiac toxicity limits its clinical application, the toxicity of the status of Chinese medicine in prevention and treatment of anthracycline anti-tumor drug in the heart of major. The hypothesis of the prevention and treatment of anthracycline heart toxicity by Warming Yang and activating blood and eliminating phlegm method was put forward according to the theory of Xinyang weakness as the basis and the mutual knot of phlegm and blood stasis as the underlying etiology and pathogenesis theory. The previous animal experiments have confirmed that the method of Warming Yang and activating blood and eliminating phlegm can obviously improve the systolic and diastolic function of CHF rat myocardium, and the clinical trial can improve the symptoms of palpitation. Based on cell experiments, we observed the characteristics of sequential changes in autophagy and the protective effects of Warming Yang, activating blood circulation and resolving phlegm on H9c2 myocardial cell injury. We also analyzed AMPK/ULK1 signaling pathway to explore its possible mechanism. On the other hand, by establishing an anthracycline rat model of cardiac toxicity, the cardiac function and the structure of the heart group were observed and the AMPK/ULK1 pathway molecules were detected. Objective to study the possible mechanism of Warming Yang, promoting blood circulation and eliminating phlegm in the treatment of anthracycline cardiotoxicity, which will provide a basis for individualized treatment of traditional Chinese medicine, from traditional subjective renewal to subjective identification and microbiological identification.

蒽环类抗肿瘤药在白血病、乳腺癌及多种恶性肿瘤均有很好的疗效,尤其在目前乳腺癌的治疗中发挥着不可替代的作用,但其严重的心脏毒性限制其临床应用，中医药防治蒽环类抗瘤药心脏的毒性地位重大。根据心阳虚弱为本，痰瘀互结为标的病因病机理论，提出温阳活血化痰法防治蒽环类心脏毒性的假说。前期动物实验已经证实温阳活血化痰法能明显改善CHF大鼠心肌收缩和舒张功能，且临床试验可以改善心悸症状。本课方面基于细胞试验，观察自噬时序性变化特点及温阳活血化痰对H9c2心肌细胞损伤的保护作用，并分析AMPK/ULK1信号通路探讨其可能的的作用机制。另一方面通过建立蒽环类心脏毒性大鼠模型，观察大鼠心功能及心脏组结构，检测AMPK/ULK1通路分子。研究温阳活血化痰法干预蒽环类心脏毒性可能的作用机制，该研究将为中医个体化治疗由传统主观辩证更新为主观辩证与微观生物学标识相结合提供依据。

蒽环类药物是一种广泛的抗肿瘤药物，临床应用广泛，但其严重的心脏毒性损伤的死亡风险已超过了癌症复发的风险。本研究基于本虚标实的病机，从“痰瘀互结”角度，提出温阳活血化痰方可防治蒽环类心脏毒性损害的假说。体内研究采用大鼠腹腔注射阿霉素，建立阿霉素心脏毒性损伤大鼠模型。体外研究采用阿霉素刺激建立阿霉素毒性H9c2心肌细胞模型。温阳活血化痰方干预后，观察组织病理形态学改变；同时进行以AMPK/ULK1-自噬-凋亡介导信号通路为切入点的机制探讨，WB检测心肌组织和H9c2细胞中LC3Ⅱ、Beclin-1、p62、pAMPK、pULK1、caspase3表达；流式细胞学检测心肌细胞凋亡情况等。结果发现温阳活血化痰方可改善大鼠心功能；减轻心肌组织病理损伤；下调pAMPK、pULK1、LC3Ⅱ，Beclin-1、caspase3蛋白表达，增加p62表达，降低H9c2心肌细胞凋亡率。表明温阳活血化痰方可防治蒽环类心脏毒性损害，其机制可能与调控AMPK/ULK1信号通路，抑制心肌细胞自噬，逆转心肌细胞凋亡机制有关。诠释中医痰瘀理论的现代病理基础，充实和完善阿霉素心脏毒性损害的中医辨证施治体系，为新药开发提供新的思路及理论依据。