基于网络靶点效应拓扑学分析的针药并用治疗脑梗塞机制研究

The combination of acupuncture and medicine obviously increases clinical effects for prevention and treatment of diseases. In past dynasties, the doctors, known drug needle and medicine, are highly recommended for the necessary qualities of their own. According to the characteristics of the combination of acupuncture with medicine, by way of principles of network topology, on the bases of previous research on treatment of cerebrovascular disease, it was pointed out the hypotheses that the acupuncture and medicine could act on the meridian and zangfu viscera, and make changes of biochemical metabolic reaction on the body to act on network targets, the final systematical effects were observed through the actions of acupuncture, medicine, and the their synergistism, and were determined availably by network target effect overlayed topological regularities”. This project will study on treating the cerebral infarction disease as a break-through point, together combining with the clinical effective treatment of “XingnaoKaiqiao” acupuncture and “Buyanghuanwu decoction”, and to carry out the investigation on the effective influences of network targets linked in energy, toxicity, oxygen free radicals, inflammatory immunity, apoptosis, vascular nerve regeneration and reconstruction, through treating the cerebral infarction disease by acupuncture, medicine, and their combination respectively, thereby to clarify the overlayed regularities of network target effect in combination of acupuncture and the Buyanghuanwu decoction as well as it’s ingredient composites, to obtain the adequate acupuncture and medicine compatibilities, in order to answer network target mechanisms in combination of the “Xingnaokaiqiao” acupuncture method and Buyanghuanwu decoction in treatment of cerebral infarction, finally to lay a foundation for establishment of the universal method how to explore the mechanisms on the combination of nacupuncture and medicine, and for guiding to remedy the clinical treatment protocols.

针药并用能明显提高临床疗效，知针知药为历代良医必备素质。根据针药并用作用特点，结合网络拓扑学原理，在前期脑血管疾病治疗研究的基础上提出了“针药都作用于经络脏腑，改变（善）机体生化代谢反应而作用网络靶点，最终体现出针、药及针药倍增（减）的综合效应，可用网络靶点效应的拓扑学叠加规律进行探测”的假说。本项目以脑梗塞的治疗为切入点，结合临床有效的“醒脑开窍”针刺法，并用补阳还五汤，进行针刺、药物及针药并用对脑梗塞能量、毒性、自由基、炎症免疫、细胞凋亡、血管神经再生与重构等网络靶点效应影响的研究，阐明针刺法并用补阳还五汤成分及其配伍对脑梗塞网络靶点的叠加规律，探讨针药并用的配伍方案，旨在回答“醒脑开窍”针刺法并用补阳还五汤治疗脑梗塞的网络靶点机制，为创立针药并用的普适方法，指导修正临床针药并用治疗脑梗塞方案奠定基础。

本项目运用生物流量和化学势平衡原理，首次建立了网络动力学平衡常数和热力学拓扑学分析方法：可由平衡常数特征值、特征向量、成分与靶点的化学势大小排序确定作用大小，并用活度系数相关系数建立拓扑网络，实现整体实验研究。运用本法先进行了脑梗塞的网络针灸学研究，从371个靶点中优选出18个靶点，并进行造模后后1天至14天网络动力学与热力学研究，确定了个体作用系数、大小和方向及整体的网络拓扑特征研究。再对补阳还五汤中的778个成分进行网络药理学、动力学研究，先筛选取68个关键靶点，并与30个入血成分进行对接研究，最终确定并进行了黄芪皂苷Ⅰ等20个成分、IL-6等20个靶点的网络动力学与热力学实验验证，排序并确定了其作用大小、方向和活度系数，绘制了活度系数相关系数的拓扑网络。从针刺和药物两方面验证了网络动力学与热力学拓扑学分析方法的可行性。再采用本法对“醒脑开窍”针刺法并用补阳还五汤及其配伍机理进行了研究，结果表明：针刺、药物与针药并用组主要作用AIF、BDNF、CHOP、FASL、P38MAPK、BCL-2、P53、PDGF-B、PLGF、TNF-α、IL-6、VEGRF等12个效应靶点，占治疗效率的80%以上（包括常数项），其中针刺、药物与针药并用组的靶点效应的排序梯度变化大，而模型组排序梯度变化少，药物组的靶点效应梯度变化与单独动力学与热力学方法学实验结果相似。针药配伍实验显示针刺内关、人中、三阴交、委中，给药补阳还五汤产生正面作用，针刺尺泽和极泉作用较弱；以针刺主穴配伍补阳还五汤得分最高，以针刺辅穴配伍补阳还五汤得分较低，针刺内关、人中、委中作用较强，辅穴尺泽和极泉作用不强且相反，补阳还五汤差于针刺主穴的作用；针刺内关、人中、三阴交、委中、尺泽和极泉，给药补阳还五汤能上调IL-6、BNDF、VEGRF、TNF-α 等靶点，下调p38MAPK、FASL、PLGR、BCL-2 等靶点，可用整体排序、网络拓扑图来表征其变化大小，揭示其治疗脑梗塞机理。因此，本项目建立的针药并用的网络动力学拓扑学研究方法将会对中医药理论现代化产生重大的学术影响。