基于线粒体能量代谢动态变化探讨桂枝甘草汤"辛甘化阳"配伍原理

Guizhi Gancao Decoction is a representative prescription of "XinGan HuaYang"compatibility. But the modern connotation of its compatibility principle is still not clear. Why does"XinGan" compatibility can "HuaYang"？What is the mechanism of "Hua" and the nature of "Yang"？Whether it is associated with mitochondrial energy metabolism? This project discusses the "life vitality" as the entry point, connects the "Yang" of Chinese medicine with the "Energy" of modern medical science and puts forward the hypothesis of "the dynamic correlation of Yang and Energy". The Decoction of Cassia trig and Liquorice and its broken prescriptions are used to observe the weight、temperature and food consumption of normal animals and model animals in normal、low and hot environment, In the administration of 5, 10 and 15 days ，as well as the Serum GLU、T3、Insulin、GC、TC、TG、Urea's content and The Skeletal and cardiac muscle's CS、 α-KGDHC、PGC-1α's liveness, COXⅣ、ATP5b mRNA's expression levels, AMP、ADP、ATP's content and the dynamic changes of relevant indexes of energy metabolism like mitochondrion ADP/O、RCR and forms，to reveal the principle of medicine compatibility of Guizhi Gancao Decoction's XinGan HuaYang and mitochondrial energy metabolism relationship，and with the purpose of providing new thought train for the research of the principle of the compatibility of TCM prescriptions' features and tastes and provide evidence for properly guiding clinical practice.

桂枝甘草汤是"辛甘化阳"配伍的代表方剂，但其配伍原理的现代内涵依然不够清晰。"辛甘"配伍为什么能"化阳"？"化"的机理和"阳"的本质究竟是什么？是否与线粒体能量代谢有关？本课题以"生命动力"为切入点，将中医"阳气"和现代医学"能量"联系起来，提出"阳气化生与能量代谢动态相关"假说。通过观察桂枝甘草汤及其拆方对正常动物、模型动物在常温、低温、高温环境下给药第5、10、15天体重、体温、摄食量；血清GLU、T3、Insulin、GC、TC、TG、Urea含量； 骨骼肌与心肌CS、α-KGDHC、PGC-1α活性，COXⅣ、ATP5b mRNA表达水平，AMP、ADP、ATP含量，线粒体ADP/O、RCR、形态等能量代谢相关指标的动态影响，揭示桂枝甘草汤"辛甘化阳"配伍原理与线粒体能量代谢的关系，为方剂性味配伍原理的内涵研究提供新的思路，为指导临床合理运用桂枝甘草汤治疗"心阳虚证"提供实验依据。

本课题通过观察桂枝甘草汤及其拆方分别对正常动物与模型动物在常温、低温、高温环境下给药第15、30天体重、摄食量，血清T3、INS、GC等含量，腓肠肌、心肌PGC1α、COXⅣ、ATP5B等活性及组织ATP含量和线粒体膜电位等能量代谢相关指标的动态影响，进而阐明桂枝甘草汤“辛甘化阳”配伍原理与线粒体能量代谢的关系。研究表明：1）与正常（空白）对照组比，桂枝甘草汤持续给药15d，可分别降低常、低温环境下正常大鼠TG、T3水平，提高常温环境大鼠心肌ATP5B、PGC1α和高温大鼠心肌COXⅣ、腓肠肌PGC1α活性，而持续给药30d，桂枝甘草汤则升高常温环境大鼠血清T3和低温环境大鼠血清GLU、心肌CS、OGDH、PGC11α含量,而降低低温环境大鼠血清T3水平；2）与模型对照组相比，桂枝甘草汤持续给药15d可降低常温急、慢性心阳虚证大鼠血清GLU、TG水平、低温急性模型大鼠心肌ATP5B、CS、OGDH、PGC1α活性和低温慢性模型大鼠体重、摄食量、血清GC含量，而升高常温急、慢性模型大鼠血清T3和低温急性模型大鼠TG水平，并上调高温慢性模型大鼠心肌ATP5B、CS、PGC1α和低温慢性模型大鼠心肌、腓肠肌COXⅣ活性，而持续给药30d，桂枝甘草汤则降低常、低温急性模型大鼠血清GC水平，下调高温慢性模型大鼠心肌、腓肠肌ATP5B、OGDH、腓肠肌PGC1α活性，而上调高温急性模型大鼠心肌OGDH、低温慢性模型大鼠腓肠肌COXⅣ、心肌PGC1α活性；3）枝甘草汤对能量代谢相关指标有较为广泛的作用，进而说明桂枝甘草汤“辛甘”配伍“化阳”的机制与线粒体能量代谢密切相关；4）从对正常动物和模型动物影响的数据分析，桂枝甘草汤对模型动物能量代谢的影响较正常动物更为广泛；4）不同温度环境作用有所差异，从对模型动物的影响分析，温度干预从大到小依次为低温、高温、常温，而从对正常动物的影响分析，则温度干预从大到小为低温、常温、高温。综上所述，本课题为方剂性味配伍原理的内涵研究提供了新思路，为临床合理治疗“心阳虚证”提供了实验依据。

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