| Aconitum carmichaeli Debx. (Fuzi) as a typical representative of "Hot property" CHM, and its objective authenticity studies emphatically aimed at the differences of Fuzi, Fuzi combined with rhizoma zingiberis (Ganjiang), and combined with "Cold property" CHM, such as rhizome of chinese goldthread (Huanglian), rheum rhabarbarum, Gypsum etc. However, the investagation on the objective authenticity of "Hot property" CHM (like Fuzi) had not been reported systematicly. This issue mainly focused on the key topics above, and the main research methods and results are blow.1 The content of six alkaloids assay of Fuzi and its similar CHMs with "Hot property" based on HPLCThis part of the studies mainly evaluated the content difference of six alkaloids of Fuzi and its similar CHMs with’"Hot property", including Fuzi from different place of origins, its processed products and the plants origining from Ranunculaceae Aconitum. And the six alkaloids were aconitine, hypaconitine, Mesaconitine, Benzoyl aconitine, Benzoyl hypaconitine and Benzoylmesaconine. The results of the content assay to six alkaloids showed that the content from Fuzi and its similar CHMs with "Hot property" differ greatly. Furthermore, the results of hierarchical cluster analysis showed that Fuzi from different place of origins, its processed products and the plants origining from Ranunculaceae Aconitum can cluster to one row efficacy which envaluates by the content of six alkaloids. The results above demonstrated that the place of origin, the processing and the origin are the cause of the differences in content of target compounds. Otherwise, the date refered can be used as the materials of "Spectum-efficency" relationship analysis.2 Established a biological thermal dynamic method based on the energy metabolism of mitochondriaThis study selected the liver mitochondria as the model which related closely to the energy metabolism. We estabolished a biological thermal dynamic method based on the energy metabolism of mitochondria, which were be used to evaluate the the differences in CHMs with "Hot property". Here we investigated the origin and the concentration of mitochondria, latering a methodological study. The experimental results showed the best options were "SD Rat, and the concentration 4.0 g/mL". Methodology study confirmed that the method precision and repeatability were conform to the requirements of the experiments.3 Study on the differences on "Hot performance" and its material basis of Fuzi from different regionsThis research mainly adopted microcalorimetry to examine the different efficiency on the metabolic activity of mitochondria isolated from rat liver of Fuzi from different regions (Sichuan Jiangyou, Yunnan Butuo, Shanxi Hanzhong, Hubei Ensi, Hunan Loudi and Guizhou Tongren). We summarized the effect of the growth of mitochondrial metabolism, under the action of different concentration of sample, mitochondrial metabolism situation occurrenced regularity changing, and when the concentration of the sample is low, samples can promote the mitochondrial metabolism, characterized by the thermodynamic parameters k, Pmax and Q and Pav rised, and Tlag, Tmax degreed. But when the concentration reached to a certain amount, the growth of the mitochondrial metabolism was restrained, showing Tlag, Tmax, the thermodynamic parameters k, Pmax and Q and Pav were characterized by reducing. Thus, samples on the growth of mitochondrial metabolism had a dual role "promote/suppression". Using the principal component analysis (PCA) for getting the major dimension, the Pmax could be used to characterize drugs promote mitochondrial metabolism efficiency. So Pmax could be used to calculate the promoting effect of mitochondria metabolic (EC50) 50%, it was concluded that different origin one to promote mitochondria generate heat intensity of the lateral root of aconite:Sichuan Jiangyou>Yunnan Butuo>Shanxi Hanzhong>Hubei Ensi> Guizhou Tongren>Hunan Loudi, EC50 were 1.86,1.96,2.12,2.48,3.23,4.41 g/mL, respectivly. These results confirmed that the "Hot property" of Fuzi from different origins were in lateral objective authenticity.CCA was used for spectrum-effect relationships among area values of 37 common peaks in the UPLC-MS fingerprints and the main thermo-kinetic parameters (Pmax) values of mitochondrial metabolism. The results were be as follows:By comparison between chromatograms of samples and the reference standard solution, we found that 16-hydroxycardiopetaline, Carmichaeline, Isotalatizidine, 14-acetykarakoline. Talatisamine. SenbusineB. SenbusineA. Neoline. Hypaconine. Mesaconine, Dehydrated benzoylmesaonitine, Benzoyl Hypaconitine, Benzoyldeoxyaconine, Benzoyl Mesaconitine, Delphinine, Benzoyl Aconitine, Mesaconitine. a total of 17 components, were the material base of the Fuzi from different origins, and the made the differences in the "Hot property"4 Study on the differences on "Hot performance" and its material basis of Fuzi and its processed productsThis research mainly adopted microcalorimetry to examine the different efficiency on the metabolic activity of mitochondria isolated from rat liver of Fuzi and its processed products (Shengfuzi, Yanfuzi, Heishunpian, Paofuzi, Paotianxiong and Baifupian). We found that the effect of the growth of mitochondrial metabolism and regularity of Fuzi and its processed products was similar with Fuzi from different regionss (except Paotianxiong and Baifupian). Using the principal component analysis (PCA) for getting the major dimension, the k and O could be used to characterize drugs promote mitochondrial metabolism efficiency. We selected Q could be used to calculate the promoting effect of mitochondria metabolic (EC50) 50%, it was concluded that Fuzi and its processed products to promote mitochondria generate heat intensity:Shengfuzi>Paotianxiong>Heishunpian>Paofuzi>Yanfuzi>Baifupian, EC50 were 0.51,1.01,1.18,1.45,1.47,1.95 g/mL, respectivly. These results confirmed that the "Hot property" of Fuzi and its processed products were in lateral objective authenticity.CCA was used for spectrum-effect relationships among area values of 37 common peaks in the UPLC-MS fingerprints and the main thermo-kinetic parameters (k and O) values of mitochondrial metabolism. The results were be as follows:By comparison between chromatograms of samples and the reference standard solution, we found that 16-hydroxycardiopetaline, Isotalatizidine, Talatisamine, SenbusineA, SenbusineB, Neoline, Fuziline, Mesaconine, Dehydrated benzoylmesaonitine, 14-anisoylneoline, Benzoyl Hypaconitine, Benzoyldeoxyaconine, Benzoyl Mesaconitine, Delphinine, Benzoyl Aconitine,14-Benzoyl-lO-OH-aconine, Mesaconitine, a total of 17 components, were the material base of the Fuzi and its processed products, and the made the differences in the "Hot property".5 Study on the differences on "Hot performance" and its material basis of the plants origining from Ranunculaceae AconitumThis research mainly adopted microcalorimetry to examine the different efficiency on the metabolic activity of mitochondria isolated from rat liver of the plants origining from Ranunculaceae Aconitum (Chuanwu, Caowu and Zueshangyizhihao). We found that the effect of the growth of mitochondrial metabolism and regularity of the plants origining from Ranunculaceae Aconitum was similar with Fuzi from different regionss (except Chuanwu). Using the principal component analysis (PCA) for getting the major dimension, the Pmax and Q could be used to characterize drugs promote mitochondrial metabolism efficiency. We selected Q could be used to calculate the promoting effect of mitochondria metabolic (EC50) 50%, it was concluded that the plants origining from Ranunculaceae Aconitum to promote mitochondria generate heat intensity:Caowu>Xueshangyizhihao>Chuanwu, EC50 were 1.10,1.61,2.31 g/mL, respectivly. These results confirmed that the "Hot property" of the plants origining from Ranunculaceae Aconitum were in lateral objective authenticity.CCA was used for spectrum-effect relationships among area values of 37 common peaks in the UPLC-MS fingerprints and the main thermo-kinetic parameters (Pmax and Q) values of mitochondrial metabolism. The results were be as follows:By comparison between chromatograms of samples and the reference standard solution, we found thatCarmichaeline, Karakolidine, Isotalatizidine, Neoline, Fuziline, dehydrated benzoylhypaonitine,14-benzoylhypaconine, Benzoyldeoxyaconine, Benzoyl Mesaconitine, Benzoyl Aconitine and Hypaconitine, 14-Benzoyl-10-OH-aconine, Foreaconitine, Mesaconitine, a total of 14 components, were the material base of the the plants origining from Ranunculaceae Aconitum, and the made the differences in the "Hot property"6 Study on the biological mechanism of "Hot property" of Fuzi and its similar CHMs based on the network pharmacologyThe main method of this study was network pharmacology. Firstly, the selected 26 target component, which were obtained from the "Spectum-efficency"analysis, were imported to link in a biological information database related targets screening group of 36. DAVID analysis software then was used to conduct the GO analysis, finally confirmed the seven related to metabolism and energy metabolism pathway and attach a subclass medicinal materials "Hot property" was most relevant, it mainly included fatty acid metabolism, polysaccharide biosynthesis, metabolism, fatty acid metabolism, membrane lipid metabolism, phospholipid metabolism and glucolipid metabolic pathways. With the above biological thermal dynamics research results have certain conformity degree, illustrates the objective authenticity of subclasses medicinal herbs’"Hot property", its biological mechanism mainly associated with energy metabolism in the body of multiple links.This study was firstly selected he rat liver mitochondria as the model, which associated with energy metabolism. We systematicly examined the influence on the energy metabolism of mitochondria of Fuzi and its similar CHMs with "Hot property’ (including Fuzi from different place of origins, its processed products and the plants origining from Ranunculaceae Aconitum.), and demonstrated that the objective authenticity was the characteristics of samples. Using the related thoughts of spectrum-effect, screened the influence with subclasses of medicinal herbs "Hot property" chemical composition, and adapted the UPLC-MS to identify chemical composition. Network pharmacology methods were used to intergretive the effective compounds and the protential biological targets to simulation the pathways which can characterize its connotation of "Hot property". It could be a nice reference to the clinical use of hot natured CHMs, and the "spectrum-efficency" relationship analysis research mode also could be a typical method to detect the material basis of the nature charicteristics of CHMs. |
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