ABSTRACT
Objective:To analyze the common active ingredients， potential target genes and pathways of Ginseng Radix et Rhizoma "Tonifying Qi" and Notoginseng Radix et Rhizoma "Enriching blood" in alleviating fatigue based on the network pharmacology technology. And the compound ingredients of total Ginsenoside Ginseng Root and Notoginseng total Saponins were selected to verify the core target genes in vitro. Method:The main active ingredients and related targets of Ginseng Radix et Rhizoma and Notoginseng Radix et Rhizoma were screened by traditional Chinese medicine systems pharmacology （TCMSP）. The data of fatigue genes were established by GeneCards comprehensive database and Human Mendelian Genetic Integrated Database（OMIM）. Depending， The data sets of fatigue-related genes are established based on the data bank of GeneCards and OMIM. The intersecting genes of drugs and disease were obtained by R software. Cytoscape software was used to establish the regulatory network among the active ingredients， drug targets and fatigue-related genes. PPI network of intersecting genes was constructed by STRING 11.0 software， and the core genes were screened by CytoHubba software and Matthews correlation coefficient （MCC） algorithm. Based on the results of network analysis， 24 male SPF ACR mice were randomly divided into control group， total Ginsenoside Ginseng Root group （0.08 g·kg-1） and Notoginseng total Saponins group （0.08 g·kg-1）. The corresponding drugs were given for 3 weeks. The expressions of core genes in muscle tissue were detected by real-time fluorescence quantitative PCR. Result:The 20 active components and 181 drug targets were screened from TCMSP. 33 intersecting genes of diseases and drugs were obtained when compared with GeneCards and OMIM comprehensive database using R software. 10 core genes including aryl hydrocarbon receptor （AHR）， androgen receptor （AR）， glutathione S-transferase P1 （GSTP1）， cysteine proteinase-3（Caspase-3）， cytochrome p450 enzyme 3A4 （CYP3A4）， intercellular adhesion molecule 1 （ICAM1） and nuclear factor kappa B inhibitor alpha （NFKBIA） were screened out by the algorithm of MCC. Total Ginsenoside Ginseng Root and Notoginseng total Saponins had no significant effect on GSTP1 and ICAM1 genes， but they could significantly inhibit the expressions of AHR， CYP3A4， Caspase-3， NFKBIA and AR （P<0.05，P<0.01）， and there were no significant difference in anti-fatigue effect between total Ginsenoside Ginseng Root and Notoginseng total Saponins groups. Conclusion:The mechanism of anti-fatigue of Ginseng Radix et Rhizoma and Notoginseng Radix et Rhizoma may be related to the regulation of AHR， CYP3A4 and Caspase-3 genes， and there is no significant difference in their anti-fatigue effects， through the analysis of network and experimental verification.
ABSTRACT
Objective:To explore the mechanism of modified Guipitang in the treatment of Yin-Fire insomnia with anxiety with the help of network pharmacological analysis technology. Method:Traditional Chinese Medicine Systems Pharmacology (TCMSP) was used to screen the main components and target genes of modified Guipitang. GeneCards and Online Mendelian Inheritance in Man (OMIM) were used to establish the target gene sets of insomnia and anxiety. STRING 11.0 software was used to analyze the interaction between the overlapping genes, and Cytoscape_3.6.1 software analysis and Matthews correlation coefficient (MCC) algorithm were used to screen the core genes. Based on the results of network analysis, 48 SD female rats were randomly divided into blank control group, model group, eszopiclone tablets group (0.2 mg·kg-1·d-1), modified Guipitang low，medium，and high-dose groups (0.31，1.25，5 g·kg-1·d-1). The model of insomnia with anxiety was established by intraperitoneal injection of Para-chlorophenylalanine (PCPA) and these rats were treated with corresponding drugs for 7 days. Then the frequency, time and distance of the activities were observed in the experiment of autonomic activity. Real-time quantitative polymerase chain reaction (PCR) was used to detect the mRNA expressions of proactivated protein kinase 8 (MAPK8), RAC-alpha serine/threonine protein kinase (Akt1), mitogen-activated protein kinase 3 (MAPK3) and interleukin-6 (IL-6) in rat hippocampus. Result:A total of 228 active compounds were screened from TCMSP database and 181 intersecting genes of diseases and drugs were obtained by comparing with GeneCards and OMIM comprehensive database. 9 core genes, including MAPK3, MAPK8, Akt1 and IL-6 were identified by STRING software and MCC algorithm. Animal experiments showed that the number of activity times, time and distance of modified Guipitang in high and medium dose groups were significantly lower than those in the model group. The high and middle dose groups of modified Guipitang could significantly inhibit the mRNA expression of MAPK3, MAPK8, Akt1 and IL-6 in hippocampus（P<0.01）, while the low dose group had no significant effect. Conclusion:The mechanism of modified Guipitang in treating Yin-fire insomnia with anxiety may be related to the regulation of MAPK3, MAPK8, Akt1 and IL-6 genes.
ABSTRACT
Zuoguiwan is a classic prescription for replenishing vital essence, tonifying kidney-Yin and nourishing the bone marrow. Zuoguiwan is one of the effective prescriptions for the prevention and treatment of osteoporosis (OP), which reflects the thought of Reinforcing Yang from Yin. The OP animal model simulates the pathological state and pathogenesis of OP in human, which is an important means to research the pathogenesis of OP and verify the effect of drugs. In this paper, two kinds of animal models and characteristics of Zuoguiwan in the prevention and treatment of osteoporosis were discussed in details. They are the primary osteoporosis animal models, including ovariectomized animal models and spontaneous elderly osteoporosis animal models, and the secondary osteoporosis models, including glucocorticoid-induced animal models, cyclophosphamide-induced animal models and subtotal nephrectomy animal models. The evaluation methods of Zuoguiwan in preventing and treating OP, including bone absorption markers and bone formation markers analyzed by enzyme-linked immunosorbent assay (ELISA), bone mineral density detected with dual-energy X-ray, the number of trabeculae, trabecular segregation, trabecular thickness, bone volume/tissue volume ratio and bone surface/volume ratio analyzed using micro-CT, bone pathological morphology observed by hematoxylin-eosin staining, bone biomechanical properties, such as the maximum load force based on biomechanical test. In order to provide scientific reference for the basic and clinical research of Zuoguiwan, the OP animal models and the pharmacodynamic effect of Zuoguiwan are evaluated comprehensively with five different and objective evaluation methods. However, the animal model of OP needs to be further optimized to highlight the pathogenesis and syndrome characteristics of Zuoguiwan in the treatment of OP.
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Objective: To investigate the mechanism of Zuoguiwan in treating ovariectomy-induced osteoporosis rats by receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) signaling pathway mediated by β2-adrenergic receptor (β2AR). Method: Forty Sprague-Dawley female rats were randomly divided into Sham-operated group (Sham) and four ovariectomized (OVX) subgroups. Rats in Sham and OVX groups were treated with 17β-estradiol (50 μg·kg-1·d-1), and low and high-dose ZGW (2.3,4.6 g·kg-1 lyophilized powder) for 3 months, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to measure the serum markers of bone turnover. Micro-CT was used to evaluate and measure trabecular bone microarchitecture and bone mineral density (BMD) of the right distal femur. Western blot analysis and Real-time PCR were used to measure mRNA and protein expressions of β2AR, OPG and RANKL. Result: After 12 weeks of treatment with Zuoguiwan, the level of serum β-cross-linked c-telopeptide of type Ι collagen (β-CTX) (PPPβ2AR in the hypothalamus (PPConclusion: The mechanism of Zuoguiwan in alleviating BMD and trabecular bone microarchitecture in ovariectomy-induced osteoporosis rats might be related to the regulation of RANKL/OPG Pathway mediated by β2AR.
ABSTRACT
Ginseng has effects in reinforcing vital energy,invigorating health effectively and relieving fatigue symptoms,and ginsenoside( GS) is the main component of its anti-fatigue effect. Totally 17 active components and 92 drug targets of ginseng compounds were screened from Traditional Chinese Medicine Systems Pharmacology; and 78 intersecting genes of diseases and drug targets were obtained based on R Language Technology. The protein-protein interaction( PPI) network was constructed by STRING 11. 0 software,and Matthews Correlation Coefficient( MCC) algorithm was used to screen core target genes. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to analyze the major genes and their roles in regulatory networks. The results indicated that ginseng could regulate the core target genes,including AKT serine/threonine kinase( AKT1),interleukin-1β,Toll-like receptor binding molecule 1( ICAM1),mitogen-activated protein kinase 8( MAPK8),AP-1 transcription factor subunit( JUN),transducer and activator of transcription 1( STAT1) and prostaglandin peroxidase synthase 2( PTGS2). It could participate in the functions of cytokine receptor binding,cell adhesion molecule binding and tumor necrosis factor receptor superfamily binding,and also regulate the signal pathways of tumor necrosis factor,interleukin 17 and c-type lectin receptor,so as to exert an anti-fatigue effect. Based on the results of network analysis,32 four-week-old male SPFACR mice were randomly divided into control group,low-dose ginsenoside group,middle-dose ginsenoside group and high-dose ginsenoside group. The corresponding drugs were administrated for 3 weeks. The results showed that GS could significantly up-regulate the expressions of STAT1 and AKT1( P<0. 01,P<0. 05),and downregulate the expressions of PTGS2 and JUN( P<0. 01). However,there was no significant effect on MAPK8,IL-1β and ICAM1. Ginseng's anti-fatigue regulation network was constructed through network pharmacology,and the results were verified by experiments,in order to reveal the anti-fatigue mechanism of ginseng and provide scientific basis for its clinical application.