[Mechanism of Alisma orientale in treating nonalcoholic fatty liver disease].

In this study, network pharmacology technology was combined with molecular docking technology and experimental verification to clarify the active ingredients, potential targets and mechanism of Alisma orientale for nonalcoholic fatty liver disease(NAFLD), providing a basis for its clinical application. The active ingredients of A. orientale were screened through traditional Chinese medicine systems pharmacology database(TCMSP), and the potential targets related to both active ingredients and NAFLD were predicted through protein databases by considering the oral bioavailability(OB) and drug-likeness(DL). The "active ingredient-potential target" network was constructed by using Cytoscape software, and the molecular docking was performed between active ingre-dients and potential targets. KEGG pathway analysis and enrichment analysis were performed through DAVID biological information annotation databases. ClueGO software was used to analyze target GO annotation. Western blot and immunocytochemistry were used to detect the protein expression levels, and fluorescent probe was used to detect the reactive oxygen species(ROS) generation level. The results revealed that 7 active ingredients of A. orientale were obtained from TCMSP database and analysis platform, 140 ingredient-related targets were screened, and 59 potential targets were obtained by intersecting disease targets with ingredient-related targets. Molecular docking showed that 7 active ingredients of A. orientale could act on the potential targets including 3-hydroxy-3-methylglutaryl-coenzyme A reductase(HMGCR) and tyrosine-protein phosphatase non-receptor type 1(PTPN1). In addition, KEGG enrichment analysis showed that the potential targets were mainly enriched in inflammatory mediator regulation, insulin resistance, neuroactive ligand-receptor interaction, vascular smooth muscle contraction, FcγR-mediated phagocytosis and other related pathways of tryptophan(TRP) channel. GO enrichment analysis showed that potential targets mainly affected the biological processes of G-protein coupled receptor signaling pathway, organic hydroxyl compound transport, positive regulation of lipid biosynthesis process, positive regulation of lipid metabolic process. Western blot, immunocytochemistry and fluorescent probe confirmed that the extract of A. orientale could reduce HMGCR and PTPN1 protein expression levels effectively, and also could reduce ROS production level of HepG2 cells. This study systematically revealed the material basis and mechanism of A. orientale in regulating NAFLD through multi-component, multi-target, and multi-pathway characteristics, which provided a theoretical basis and scientific basis for the clinical application of A. orientale.

Characterization of oil/water partition coefficient of chishao terpene glucoside components based on contribution rate of representative components for myocardial ischemia / 中国中药杂志

The components of traditional Chinese medicine(TCMCs) are the basic unit of raw materials for Chinese medicines, and their physical and chemical properties directly affect the choice of dosage forms and the optimization of prescriptions. However, most of TCMCs are multi-component complex systems, and the characterization of their overall properties is still in the exploration stage. On the basis of biological activity, the representative components are determined, and then the individual characteristics are fitted with the weight coefficient of efficacy contribution rate, which may provide reference for characterizing the overall properties of TCMCs. In this study, with the pharmacological effects of isoproterenol(ISO)-induced myocardial ischemia in rats as the indicators, the pharmacodynamic contribution rates of three representative components of chishao terpene glucoside components(CSTGCs) were evaluated by the normalization weighting method. The contribution rates of paeoniflorin, paeoniflorin and benzoylpaeoniflorin were 54.87%, 32.46% and 12.67%, respectively. The oil-water partition coefficients of paeoniflorin, albiflorin, benzoylpaeoniflorin in water and buffer solutions with different pH values were measured, and the oil-water partition coefficients of CSTGCs were characterized by the weight of their pharmacodynamics contribution rate. The results showed that the apparent oil-water partition coefficient(log P) of CSTGCs in the phosphate buffer system such as n-octanol-water(pH 2.0, 2.5, 5.0, 5.8, 6.8) were 0.18-0.22, indicating that CSTGCs have common absorption and low permeability, providing basis for the preparation of CSTGCs.

Characterization analysis technique of Panax notoginseng saponin subcomponents for prevention and treatment of myocardial ischemia based on their structure and effect differences / 中国中药杂志

According to the structure and effect differences of Panax notoginseng saponin components(PNSC), subcomponent division and network pharmacological characterization were conducted to provide a research basis for the medicinal properties of P.notoginseng saponin subcomponents and the technical design of unit preparations. PNSC were screened by the TCMSP database and subcomponents were classified according to systematic clustering. Then the subcomponents obtained were subjected to target prediction and attribution analysis by PharmMapper server, GeneCards, DisGeNET and HOME-NCBI-GENE database. A subcomponent target interaction network was constructed by using the STRING database. KEGG and GO enrichment analysis were performed on each subcomponent target using the DAVID database. The subcomponents-targets-pathways visualization network was constructed by Cytoscape. The subcomponent targets and pathways involved were compared to analyze the differences in anti-myocardial ischemic drug mechanisms and the rationality of subcomponent division. Eighteen compounds of PNSC were screened out, and classified into three subcomponents A, B, and C according to their properties, involving 67 targets and 17 common anti-myocardial ischemic pathways directly or indirectly related to myocardial ischemia. Subcomponent A had the highest number of targets and the target interaction was dense, possibly indicating its key role in the mechanism of pharmacodynamics. Subcomponents A, B, and C had similar basic structures, and KEGG and GO analysis showed that they all can enhance the heart function and protection of cardiomyocytes by inhibiting apoptosis, promoting angiogenesis and regulating inflammatory response to play the effect on myocardial ischemia. This study fully reflected the differences in the efficacy of various subcomponents in preventing and treating myocardial ischemia due to the different physical properties of P. notoginseng saponin subcomponents. To some extent, the differences in the efficacy of each subcomponent in the prevention and treatment of myocardial ischemia could verify the rationality of the division of P. notoginseng saponin subcomponents according to the structural properties, realizing the characterization of P. notoginseng saponin subcomponents based on structure and effect differences.

Screening of active anti-myocardial ischemia components of Panax notoginseng based on molecular docking technology / 中国中药杂志

The molecular docking technology was used in this study to virtually screen the active anti-myocardial ischemic components in Panax notoginseng, clarify the compositions of the anti-myocardial ischemic component unit and the basis for pharmacological activity of P. notoginseng, and provide the basis for the acquisition of the component raw materials and the formulation design before the preparations. One hundred and nineteen compounds in P. notoginseng were collected by searching TCMSP to establish the ligand database, and TNF, IL1 B, NFKBIA, and NOS3 which were related with myocardial ischemia were selected to create the receptor database. Then Discovery Studio software LibDock module was used to dock the ligands and receptors, with the approved small-molecule drugs which were related to targets or the treatment of myocardial ischemia disease in the DrugBank as the reference, and the average scores of approved small-molecule drugs were set as the threshold. A total of 13 compounds with a score above the threshold and in the top ranking were virtually screened. The study showed that all the 13 components screened out were saponins, which constituted the main component unit of the anti-myocardial ischemic activity of P. notoginseng, namely the P. notoginseng saponin components. After the comparative analysis of the main active residues of the approved commercial drugs and P. notoginseng saponin components on each target, the similarity of their effects suggested that the P. notoginseng saponin components may have the same anti-myocardial ischemic efficacy as clinical drugs. The components of P. notoginseng which exerted anti-myocardial ischemic activity were mainly the saponin components. The preliminary screening of the active anti-myocardial ischemic components of P. notoginseng had been completed, which provided a certain reference for the development of anti-myocardial ischemic Chinese medicine component preparations.

Characterization of overall biopharmaceutical properties of traditional Chinese medicine components based on identification of representational components / 中国中药杂志

Preformulation research, an in-depth research for pharmacological and pharmaceutical properties of raw materials, has been widely used in the field of chemical drugs. Although traditional Chinese medicine components (TCMCs) are the basic units in Chinese medicine preparation, the properties characterization of these components is still in an exploratory stage, because empiricism and blindness are still present in the development of the Chinese medicine preparations. TCMCs, a complex multi-component system, is very difficult to be analyzed in details. Herein, a research idea is put forward for the characterization of overall properties by using representative compositions. Firstly, various composition groups were set up for screening the representative components by in vitro and in vivo efficacy evaluation according to the original proportion. Then the equilibrium solubility, oil/water (O/W) partition coefficient and apparent permeability coefficient of the representative components were detected. Subsequently, the similarity assessment and discrete analysis were performed for the subdivision of TCMCs. The overall properties of TCMCs were fitted by mass fraction or efficacy contribution of each representative component, so as to characterize the overall properties of components/sub-components.

Discovery and verification of representative ingredients of Chishao terpene glucoside components against ischemia and hypoxia injury / 药学学报

The discovery and verification of components are prerequisites for developing of component preparations. The molecular docking technique and pharmacodynamic activity evaluation provide effective methods for the discovery and verification of the representative components of Chishao terpene glucoside components (CSTGCS) against ischemia and hypoxia injury. The chemical constituents of CSTGCS were analyzed qualitatively by UPLC-TOF/MS/MS. Main chemical constituents were docked with key receptor proteins of myocardial ischemia to preliminarily screen anti-ischemia active ingredients, and screening for main active ingredients with Libdockscore. Then a H9c2 cell hypoxia injury model was established, and creatine kinase (CK), lactate dehydrogenase (LDH), superoxide dismutase (SOD), malondialdehyde (MDA) were determined to screen the representative combinations in CSTGCS. In addition, apoptosis index, apoptotic protein expression and mitochondria-associated mRNA levels were determined to verify the inhibition of the representative components (RCS) on the apoptosis of hypoxic cells. Eventually, the representative components of CSTGCS were determined. The results showed that paeoniflorin, albiflorin, benzoyl paeoniflorin and oxypaeoniflorin were considered to be the main active components because of their high matching with target proteins (4TWT, 3O4O, 4KZN, 1M9J) in space and energy. There was no statistical difference in regulating CK, LDH, SOD, MDA levels and maintaining mitochondrial function as well as inhibiting cell apoptosis between CSTGCS group and RCS group (paeoniflorin + albiflorin + benzoyl paeoniflorin combination). Therefore, paeoniflorin, albiflorin and benzoyl paeoniflorin were selected as the most representative ingredients of CSTGCS against ischemia and hypoxia injury, providing a basis for the overall properties of the components and formulation of CSTGCS.