| Formula is the basic form of traditional Chinese medicine in clinical, the compatibility is core of formula, chief、adjuvant、assistant and courier principle is the major method for the compatibility judgement, what is more, action mechanism of formula is the synergy between Multi-components and multi-targets. However, the effective components are not clear, targets are unknown, which seriously hindered the action mechanism of formula research progress."Lord sick that the king," which explained the relationship between the medcine in the formula and the disease, but also explain that the formula compatibility always change according to the disease. Presently,The main method based on reductionism study can not fully and scientificly explain the formula computability law and action mechanism. So the exploration of effective chemical components and targets is still important for the study of formula. Gegen Qinlian Decoction (GGQLD),first recorded in the book of<shanghanlun>, is a classic formula, which was consisted of four traditional Chinese medicine herbs, Pueraria lobata(Wild) Ohwi (PO); Scuterllaria baicalensis Georgi(SG);Coptis chinensis Franch(CF);Glycyrrhiza uralensis Fisch(GF)。Now,it was wildly used to treated type2diabete metllius. This paper aims to integrate the technologies of uniform design, mathematical statistics, transcriptomics, metabolomics and molecular docking to explore the compatibility law of Gegen Qinlian Decoction in the treatment of type2diabetes mellitus, and find out the effective components and targets. It will be helpful for the clinical and experiment study on Gegen Qinlian decoction1%The Quality Control of PO> SG^CFandGF Decoction Pieces. Objective:to choose better Decoction Pieces for the Gegen Qinlian Decoction Method:PO、SG、CFand GF Decoction pieces which provided by8different manufacturers was bought, then HPLC was used to control the quality of Decoction Pieces according to the requirements of Pharmacopoeia, further fingerprint was applied.Results:By contrast the main component content and water content, we found that the Decoction Pieces from8manufacturers are different in quality. Ge Gen (Henan,912014), SG(Beijing,911002), CF(Sichuan,905024), GF (Inner Mongolia, 910018), made by Yanjing sliced medicinal factory,are better than that made by other factory. The water content Decoction Pieces is8.9%,8%,7.7%,8.8%,respectively. which was lower than the moisture ruled by the Pharmacopoeia. The main components of puerarin, baicalin, berberine,(epiberberine, Huang Lianjian, BA Martin), glycyrrhizic acid, glycyrrhizin respectively is3.05%,13.9%,6.16%,3.3%,1.58%,0.678%, were higher than the standard ruled by the Pharmacopoeia, and the fingerprint of Decoction Pieces from different manufacturers are similar. Conclusion:Ge Gen (Henan,912014), SG(Beijing,911002), CF(Sichuan,905024), GF (Inner Mongolia,910018),made by Yanjing sliced medicinal factory, have better quality,which can be used for the following experiment.2、Experiment study on the compatibility regularity of Gegen Qinlian Decoction in the treatment of type2diabetes mellitus based on uniform design Objective:To analyze the compatibility law of Gegen Qinlian Decoction in the treatment of type2diabetes.Methods:In order to copy type2diabetic rat model,312rats were fed with high-fat and high-sugar diet for a month, then injected with Streptozotocin(STZ,25mg/kg) via tail vein. Only rats with FBG value were higher than11.1m M in both detection, were considered as rats with type2diabetic mellitus. The uniform design table was used to design15Gegen Qinlian Decoction group with different ratios of PO SG CFand GF. Except the control group,the rats with type2diabetic mellitus was divideded into18group, namely Gegen Qinlian Decoction1-5group, Gegen Qinlian Decoction group, type2diabetic mellitus group, Metformin group.Then the19group rats were feed with corresponding drugs for90days.90days later, rats were executed. Blood was collected and prepared of serum. The FBG, Insulin, GSP, TC, TG, HDL-C, LDL-C in the serum were detected. the Compatibility law of the treatment of type2diabetes was analysed with correlation analysis, stepwise regression and partial least squares regression analysisResults:The amount of PO in Gegen Qinlian Decoction was significantly negatively correlated with the content of FBG, TC, TG, HDL-C, LDL-C; the stepwise regression analysis result shows that the main medcine of Gegen Qinlian Decoction is PO,within the experimental range, Gegen Qinlian Decoction14is the best formula in treatment of type2diabetes; PLS results showed that accompany with the increasing ofPO、CF, and decreasing of SG、GF, the FBG, TC, LDL-C in the serum of type2diabete mettlius model rats will drop, on the basis of VIP, the order is PO>SG>CF>GF. Conclusions:Based on the relevance between herbs and type2diabetes, the compatibility law of Gegen Qinlian Decoction is that PO plays the role of chief*CFplays the role of adjuvant SG and gancao plays the role of assistant and courier.3、The study on the action mechanism of Gegen Qinlian Decoction in the treatment of type2diabetes based on metabonomicsObjective:To explore the molecular mechanism and therapeutic targets of Gegen Qinlian Decoction in the treatment of type2diabetes by metabonomics. Methods:rat molding, grouping, administration, index selection and detection method have shown chapter two methods section. By comparing the efficacy and time-effect analysis, we confirm the Gegen Qinlian Decoction14is the best formula in treatment of type2diabetic rats. Plasma was collected, Then three times methanol was added to get rid of protein, LC-MS-QTOF was applied for data acquisition, principal component analysis (PCA) and mathematical statistics was used to analyse the data.Results:After three months of administration, Gegen Qinlian14significantly lowered FBG, Insulin, HOMA-IR, GSP, TC, TG, LDL-C levels in serum of type2diabetic rat. insulin resistance status of type2diabetic rats was Changed, leucine and sphingosine-1-phosphate increased in plasma of diabetic model group by mathematical statistical methods, Gegen Qinlian14significantly decreased the content of them.Conclusion:leucine may be an biomarker for Gegen Qinlian Decoction in the treatment of type2diabetes mellitus, PPARy may be one of most important targets for Gegen Qinlian Decoction in the treatment of type2diabetes mellitus.4、The study on the action mechanism of Gegen Qinlian Decoction in the treatment of type2diabetes mellitus based on the TranscriptomicsObjective:To explore the molecular mechanism and therapeutic targets of Gegen Qinlian Decoction in the treatment of type2diabetes. Methods:rat molding, grouping, administration, index selection and detection method have shown chapter two methods section. By comparing the Pharmacodynamics and time-effect analysis, we confirm the Gegen Qinlian Decoction14is the best formula in treatment of type2diabetic rats, the liver tissue was collected for the research of the difference spectrum transcription by expression microarray, gene with expression value exceed more than two times were validated by realtime PCR and insulin resistance cell model of human liver. the expression of important hepatic gene PPARa, CEBPa, and SREBP-1C rise up, PI3K signal transduction pathway mediated by the action of IRS2was repaired, which was further confirmed in human hepatocytes in vitro model of IR.Conclusion:through IRS2/PI3K signaling pathway, Gegen Qinlian Decoction regulates glucose and lipid metabolism to Intervene IR in the liver and improve treatment of T2DM, PPARa is an important nodes of its regulation5-. Active substance prediction for Gegen Qinlian Decoction in the treatment of type2diabetes mellitus by the molecular docking technology Objective:To predict the active substance of Gegen Qinlian Decoction in the treatment of type2diabetes by molecular docking technology.Methods:ligands are the targets found by transcriptomics and metabolomics experiments, the crystal structure of them were downloaded from the protein database. before docking, the ligand and the water were extracted and receptor were treated with hydrogenation. Chemical components and structures were downloaded from Chinese herbal ingredients database and pubmed compound database in.sdf Format, sybyl2.5software was used for molecular dockingResults:flavonoids and alkaloids from Gegen Qinlian Decoction has a good capacity to agnoist PPARa and PPARy. Conclusion:flavonoids and alkaloids may be the active components for Gegen Qinlian Decoction in the treatment of type2diabetes. |
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