基于miR-30a靶向Beclin1调控自噬-凋亡探讨参麦方干预蒽环类药物心脏毒性的效应机制

Anthracycline is the cornerstone of chemotherapy for malignant tumors, and cardiac toxicity is the most common and most serious adverse reaction of anthracycline. Anthracycline heart toxicity often causes patients not to die of cancer but to die from heart complications, which has become a "heart disease" of oncologist, and its prevention and treatment is still in the exploratory stage. Studies have shown that the cardiac toxicity of anthracycline is closely related to autophagy and apoptosis of the myocardium. Beclin1 is central to the regulation of autophagy-apoptotic reticular signaling, whereas miR-30a inhibits transcription by targeting the 3'-UTR of Beclin1. Therefore, the regulation of autophagy-apoptosis based on miR-30a targeting Beclin1 may provide new targets and strategies for the treatment of anthracycline cardiotoxicity. In this study, the cardiotoxicity of anthracyclines was used as an entry point. In view of the characteristics of TCM syndrome of Qi and Yin deficiency of anthracycline drugs, we choose Shen-Mai formula which is provided nourishing Yin fluid function to intervene. Through in vivo and in vitro experiments, we want to clarify that Shen-Mai formula can exert myocardial protective effects by regulating autophagy-apoptosis. In addition, based on the pathway of miR-30a/Beclin1, we also want to elucidate the target and mechanism of Shen-Mai formula intervening anthracycline cardiotoxicity, and interpret the scientific connotation of Chinese medicine intervening anthracycline cardiotoxicity.

蒽环类药物是恶性肿瘤化疗的基石性药物，而心脏毒性是蒽环类药物最常见、最严重的不良反应。蒽环类药物心脏毒性常导致患者不是死于肿瘤而是死于心脏并发症，已成为肿瘤科医生的“心病”，其预防和治疗，还处于探索阶段。研究显示，蒽环类药物心脏毒性与心肌自噬和凋亡密切相关。Beclin1位于自噬-凋亡网状信号调控的中心环节，而miR-30a以Beclin1的3’-UTR为靶点抑制其转录。因此，基于miR-30a靶向Beclin1调控自噬-凋亡可能为治疗蒽环类药物心脏毒性提供新的靶点和策略。本项目以蒽环类药物心脏毒性为切入点，针对蒽环类药物心脏毒性的气阴两虚证候特点，选用益气固脱、养阴生津之功效的参麦方干预，通过在体与离体实验，明确参麦方调控自噬-凋亡的心肌保护效应，阐明参麦方通过miR-30a/Beclin1调控自噬-凋亡干预蒽环类药物心脏毒性的效应靶点和机制，诠释中药干预蒽环类药物心脏毒性的科学内涵。

心脏毒性是抗肿瘤药物最主要的不良反应，也是肿瘤存活者死亡率增高的首要原因，直接影响了抗肿瘤药物的疗效。心脏毒性可见于蒽环类、烷化剂类、抗代谢药、抗微管药物、靶向药物等抗肿瘤药物，其中蒽环类药物心脏毒性最为常见。蒽环类药物是化疗的基石性药物，然而其心脏毒性也是最常见、最严重的不良反应。如何在使用蒽环类药物的同时，预防或降低其心脏毒性，是心血管专家和肿瘤专家需要共同面对的问题。本项目回顾性分析了蒽环类药物心脏毒性患者的临床特征和证候分布，基于文献梳理了中医药治疗抗肿瘤药物心脏毒性的用药特点和配伍规律，基于网络药理学探讨了参麦注射液干预蒽环类药物心脏毒性的潜在机制。通过尾静脉注射多柔比星建立心脏毒性大鼠模型，结果显示参麦注射液可以减小LVEDD、LVESD，增加LVEF、LVFS，降低血清CK、LDH、cTnT、NT-proBNP、sST2、GDF-15水平，减少心肌I型和III型胶原蛋白表达，通过增加miR-30a表达水平，抑制Beclin 1 mRNA表达，降低LC3-Ⅱ/LC3-Ⅰ、Bax、Caspase-9、Caspase-3蛋白表达，增加p62、Bcl-2蛋白表达，提示参麦注射液可以减轻蒽环类药物的心肌损伤，其机制与抑制心肌自噬和凋亡相关。进一步采用多柔比星构建心肌细胞损伤体外模型，结合抑制miR-30a表达、过表达Beclin-1，结果显示参麦注射液可以提高受损心肌细胞活力，降低LDH漏出率、CK水平，抑制心肌细胞自噬流，降低Beclin-1、LC3-Ⅱ、LC3-Ⅱ/LC3-Ⅰ蛋白表达，增加p62蛋白表达；抑制心肌细胞凋亡水平，上调Bcl2表达，下调Bax、Cleaved-Caspase-9、Cleaved-Caspase-9/Caspase-9、Cleaved-Caspase-3、Cleaved-Caspase-3/Caspase-3表达，提示参麦注射液通过干预miR-30a/Beclin-1调控自噬-凋亡发挥心肌保护效应。本项目以蒽环类药物心脏毒性为切入点，针对蒽环类药物心脏毒性的气阴两虚证候特点，选用益气固脱、养阴生津之功效的参麦注射液干预，通过在体与离体实验，明确了参麦注射液调控自噬-凋亡的心肌保护效应，基于miR-30a/Beclin-1阐明了参麦注射液调控自噬-凋亡干预多柔比星心脏毒性的效应机制，诠释了中药干预蒽环类药物心脏毒性的科学内涵。