基于MAPK和NF-κB信号通路研究四逆汤抗炎心肌保护作用的分子机制

Despite the long clinical history of Sini decoction on treating cardiovascular diseases, the molecular mechanisms of its cardioprotective effects has not yet been clarified. There is an increasingly common view that the signal molecules of myocardial injury contribute to pathological processes of cardiovascular diseases through initiating inflammatory responses via MAPK and NF-κB signaling pathways. In exciting preliminary data, our project team found that Sini decoction could ameliorate the extent of myocardial inflammatory injury induced by Isoproterenol through NF-κB signaling pathway. Moreover, through HPLC method, we detected some active compounds such as glycyrrhizinic acid and 6-gingerol in Sini decoction. These active compounds have both cardioprotective effects through regulating NF-κB signaling pathway and anti-inflammatory effects through regulating MAPK and NF-κB signaling pathways. Based on the information above, we hypothesize that Sini decoction can protect myocardium through inhibiting inflammatory responses via MAPK and NF-κB signaling pathways. The scientific hypothesis will be authenticated by measuring the changes of biomarkers of myocardial injury and inflammation and related protein expressions of MAPK and NF-κB signaling pathway through modern cytological and molecular biology analysis when MAPK and NF-κB signal pathways are inhibited by inhibitors or siRNA after Sini decoction pretreating in vivo and in vitro models of myocardial injury and inflammation. Moreover, the molecular mechanism correlation between Sini decoction and anti-inflammatory compounds (glycyrrhizinic acid or 6-gingerol) will be observed after these compounds pretreating models through the same research methods, which will clarify the hypothesis from the perspective of material basis. If the hypothesis is validated, the project will disclose the molecular mechanisms of cardiovascular effects from the perspective of anti-inflammation and provide an important scientific evidence for clinical applications of Sini decoction. Furthermore, the project will give us a research clue on pharmacodynamic material basis of Sini decoction.

四逆汤临床应用历史悠久，但其心肌保护作用的分子机制仍未完全阐明。心肌损伤信号分子可通过MAPK和NF-κB信号通路激活炎症参与心血管疾病病理过程已获共识。前期研究发现四逆汤减轻心肌炎性损伤的机制与NF-κB信号通路有关；HPLC检测出四逆汤中含既可通过MAPK和NF-κB通路进行抗炎又可保护心肌的物质——甘草酸和6-姜酚。据此，项目组推测四逆汤可通过MAPK和NF-κB信号通路发挥抗炎心肌保护作用；拟通过四逆汤干预心肌损伤和炎症的体内、体外模型，并采用抑制剂与siRNA阻断相关模型的信号通路，运用现代细胞分子生物学的技术与方法检测心肌损伤和炎症标志物、通路相关蛋白表达等验证假说；在此基础上，通过对甘草酸、6-姜酚进行与假说内容相关的研究，进一步从物质基础层面诠释假说。本项目将从抗炎角度揭示四逆汤心肌保护作用的分子机制并为拓展其临床应用提供理论依据，也为四逆汤药效物质基础研究提供借鉴意义。

四逆汤临床应用历史悠久，但其药理作用机制尚未完全阐明。近年来，炎症参与心血管疾病的发生发展过程已得到广泛认可。本项目运用现代细胞与分子生物学技术通过建立脂多糖诱导RAW264.7细胞炎症模型和脂多糖诱导小鼠C57BL/6炎症模型观察到四逆汤可调控IL-6和IL-1β等炎症因子来发挥抗炎作用，亦通过建立过氧化氢诱导H9C2细胞损伤模型和异丙肾上腺素诱导SD大鼠心肌损伤模型观察到四逆汤具有心肌保护作用；而这些作用的分子机制均与MAPK和NF-κB信号通路有关。此外，我们还找到与四逆汤抗炎作用密切相关的其中一种单体——6-姜酚。本项目从调控MAPK和NF-κB信号通路角度揭示了四逆汤抗炎心肌保护作用的分子机制，为四逆汤临床应用提供了科学数据支撑，也为四逆汤药效物质研究提供了研究参考。