中文摘要
胸痹的基本病机是气虚血瘀，治疗则以补益心气，活血化瘀为法。现代医学研究表明氧化应激性损伤与心肌疾病的发病机理密切相关，氧化应激可直接或间接诱导心肌细胞凋亡。本课题组前期研究以脑心通方为益气活血方的经典代表，首次发现CTRP家族蛋白CTRP2可在心肌细胞株H9c2中表达，脑心通方通过抗氧化作用、促进CTRP2 的表达、激活p42/44 MAPK、影响胞浆钙离子流动和线粒体膜电位保护氧化应激的心肌细胞。CTRP2与p42/44 MAPK在心肌氧化应激时的关联，以及炎症因子TNF-α对CTRP2的调控机制，目前国内外尚无报道。本课题拟从细胞及在体水平进行系统研究，利用RT-qPCR、SiRNA 基因沉默及基因过表达等技术探讨TNF-α/CTRP2/p42/44 MAPK信号通路在心肌氧化应激过程中的作用，以及脑心通方通过该信号通路发挥心肌保护作用的具体机制，为氧化应激性心肌损伤的防治提供新靶点。
英文摘要
The basic pathogenesis of chest-Bi syndrome is qi deficiency and blood stasis, and the usual treatment method is to tonify heart-qi and activate blood stasis. The modern medicine research indicates that myocardial injury in the pathogenesis of myocardial diseases is closely related to oxidative stress which may directly or indirectly induce myocardial cells apoptosis. Naoxintong recipe is a representative formula of Yiqihuoxue prescription to treat ischemic cardiovascular diseases, and our previous studies found that CTRP2 was expressed in H9c2 cell line for the first time, and Naoxintong played a protective effect in H9c2 cells by antioxidation, increasing the expression of CTRP2, activating p42/44 MAPK, and influencing the mitochondrial membrane potential and concentration of intracellular calcium. There are no reports at home and abroad about the relationship between CTRP2 and p42/44 MAPK, and the regulatory mechanism of the inflammatory cytokine TNF-α on the expression of CTRP2. This research will focus on the mechanism of TNF-α/CTRP2/p42/44 MAPK signaling pathway involved in myocardial oxidative stress, and the specific protection mechanism of Naoxingtong through TNF-α/CTRP2/p42/44 MAPK signaling pathway in vitro and in vivo experiments. This research subject would provide new targets for the prevention and treatment of myocardial injury induced by oxidative stress, and establish a theoretical basis for clinical application of Yiqihuoxue prescription, which has important theoretical significance and application value.
结题摘要
胸痹的基本病机是气虚血瘀，治疗则以补益心气，活血化瘀为法。现代医学研究表明氧化应激性损伤与心肌疾病的发病机理密切相关，氧化应激可直接或间接诱导心肌细胞凋亡。本课题组前期研究以脑心通方为益气活血方的经典代表，首次发现CTRP家族蛋白CTRP2可在心肌细胞株H9c2中表达，脑心通方通过抗氧化作用、促进CTRP2 的表达、激活p42/44 MAPK、影响胞浆钙离子流动和线粒体膜电位保护氧化应激的心肌细胞。CTRP2与p42/44 MAPK在心肌氧化应激时的关联，以及炎症因子TNF-α对CTRP2的调控机制，目前国内外尚无报道。本课题拟从细胞及在体水平进行系统研究，利用RT-qPCR、SiRNA 基因沉默及基因过表达等技术探讨TNF-α/CTRP2/p42/44 MAPK信号通路在心肌氧化应激过程中的作用，以及脑心通方通过该信号通路发挥心肌保护作用的具体机制，为氧化应激性心肌损伤的防治提供新靶点。研究结果表明建立过氧化氢损伤H9c2心肌细胞株模型以及急性心梗大鼠动物模型，从细胞和动物实验证实氧化应激产生的炎症因子 TNF-α可调控下游细胞因子 CTRP2 的表达，CTRP2通过介导 p42/44 MAPK 的磷酸化，参与心肌损伤过程。当心肌受到氧化应激损伤时，CTRP2蛋白表达降低，炎症因子TNF-α升高，p42/44 MAPK表达增多，心肌细胞凋亡率增加，氧化应激相关分子表达升高，例如细胞内活性氧产生增多，使用氧化应激抑制剂α-硫辛酸可抑制此过程，说明TNF-α/CTRP2/ p42/44 MAPK 通路可参与心肌氧化应激性损伤的发病机制。此外，从细胞和动物水平证实脑心通方可调控 TNF-α/CTRP2/p42/44 MAPK 通路发挥心肌保护作用，逆转病理过程，抑制氧化应激，降低心肌细胞凋亡率，对急性心梗大鼠和过氧化氢损伤心肌细胞株具有保护作用。本研究为多种心肌疾病的防治和药物研发提供新思路、新靶点和新的科学理论依据和实验证据，同时考察和丰富中药脑心通方心肌保护作用的机制，确证“益气活血”的现代分子生物学依据，为心肌疾病的预防和治疗提供新的治疗策略。