| This study used whole gene chips for the study of meridian theory of traditional Chinese medicine, focusing on Gastrodia and Uncaria which is the classic therapy for Liver-Yang. Gene expression level can qualitatively and quantitatively reflect the full view of pharmacological mechanism.The Compatibility of Gastrodia and Uncaria is referred to the clinical application of Tianma Gouteng Decoctiont.Objective:1. To definite the tissue which the herbal attributing to liver meridian mainly effect on, and to look for relationship of Chinese "organs" and the anatomy of organs;2. To analyze the interaction manner between Gastrodia and Uncaria during the compatibility process;3. To investigate the important biological pathways of the herbal attributing to liver meridian;4. To clarify the main mechanism of "Liver attribution of Gastrodia".Methods:1. Theoretic researchThe relevant literatures about modeling of Liver-Yang were reviewed. SHR rats were determined as the model of "Liver-Yang" which is the closest to the ideal condition. What's more, this modeling approach had no interference for herbal study.Through the study of bio-informatics, the differential gene expression spectrum analysis method was set up., Relevant literatures about the application of gene chip in Modernization of Traditional Chinese Medicine were reviewed to established the Union of gene chip analysis techniques and traditional Chinese medicine research- "disease-drug treatment" differential expression profile.2. Experiment researchFour parallel control groups were set up including blank group, Gastrodia treatment group, Uncaria treatment group, Gastrodia and Uncaria combined decoction treatment group. SHR rats were continuously administered intragastrically for 10 days then were killed 30 minutes after the last administration. Histopathological slides of four rats groups were produced, including heart, liver, pancreas, lung, kidney, brain, and major blood vessel, to observe whether drugs reversed tissue lesions effectively; Total RNAs from liver and kidney of four rats groups were extracted by liquid nitrogen grinding,8 samples in total were hybridized with Agilent whole mouse genome chip after Cy3 labeling. Final data were analyzed to establish "disease-drug treatment" differential expression profile, filterting target genes, carrying on cluster analyze, biological function (GO) analyze and biological pathway analyze.Results:1. Compared with blank group, SHR rats from the glomerular hyalinization of Gastrodia treatment group, Uncaria treatment group, Gastrodia and Uncaria combined decoction treatment group were improved significantly. Renal and hepatic artery vascular wall thickening situation were reversed. The liver cells of Uncaria treatment group rats vacuolar degenerated significantly, and vesicles appeared individually in the liver of this group.2. The genes were differentially expressed in 8 samples as shown in Scatter (Appendix Figure 2-1-Appendix Figure 2-1). Totally,30 Gastrodia-kidney related genes,26 Uncaria-kidney related genes,16 Gastrodia-liver related and 15 Uncaria-liver related genes were selected.3. Cluster analysis results were shown in Figure 5-5.Conclusions:1. Liver and Kidney were the certain tissues that Gastrodia and Uncaria mainly effect on.2. During the compatibility process, Gastrodia acted as master, meanwhile, Uncaria acted as slave. The compatibility makes the pharmacological effects more focused on the liver, consistent with " Guiding compatibility" theory.3. Liver-yang was related to lipid metabolism, amino acid metabolism, G protein-coupled receptor-mediated signaling pathway disorder and imbalance in the body fluids. There also exists systemic inflammatory response.4. For patients with liver yang, Gastrodia can adjust lipid metabolism, affect high-density lipoprotein (HDL) metabolism, get involved in cholesterol ester transport, have the efficacy of anti-atherosclerosis. Gastrodia can inhibit liver inflammation; is able to adjust amino acid metabolism in order to protect the liver; is able to regulate G-protein coupled receptor mediated signal transduction pathway effectively-especially the cyclic adenosine monophosphate (AC) pathway, then the blood pressure could be decreased; can reduce dopamine (DA) and to Norepinephrine (NE) levels, improve insulin resistance, inhibit hyperthyroidism in order to calm the Liver-yang; can reduce serum total bile acid content, control of blood pressure, protect the target organs.5. Compared to single Gastrodia, the pharmacological effects on kidney of combined decoction were weaker, while the protecting for liver was more comprehensive and more significant, which means it is specially localized on liver.6. Organic cation transporter protein was involved in the transport of Gastrodia active ingredients, but the active ingredients of Gastrodia were not existed within metal ion complexes.7. Single Uncaria may have certain toxicity to the SHR rat's liver, which might cause liver disease. After administered intragastrically with Uncaria, three particular genes of SHR rats were differentially expressed:fibroblast growth factor(FGF21) expression was decreased, asparagine synthetase (ASNS) expression was decreased, soluble galectin-binding protein (LGALS3BP) expression was significantly reduced. These may be the main reasons which cause the liver function decreasing or liver cancer. Meanwhile the combined decoction can make an effective reversal of the expression of the three particular genes, which could relieve the toxicity of Uncaria.
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