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BACKGROUND:Our previous studies have confirmed that H2O2 and ginsenoside Rg1 can cause changes in reactive oxygen species levels in human periodontal ligament cells,but the correlation of reactive oxygen species with apoptosis and autophagy remains unclear. OBJECTIVE:To explore the effect and mechanism of ginsenoside Rg1 on H2O2-induced apoptosis and autophagy of human periodontal ligament cells. METHODS:Human periodontal ligament cells were divided into control group,H2O2 group and H2O2+Rg1 group.Ginsenoside Rg1(50 μmol/L)was pre-incubated for 24 hours and H2O2 was treated for 2 hours(500 μmol/L).CCK-8 assay was used to detect the proliferation ability of the cells.A fluorescent probe DCFH-DA was used to detect the reactive oxygen species level of the cells.qRT-PCR and western blot assay were used to detect heme oxygenase 1,apoptosis-related factor Caspase-3,Bax,anti-apoptotic factor Bcl-2,autophagy Beclin-1,P62,LC3,pathway-related factors phosphatidylinositol-3-kinase(PI3K),protein kinase B(AKT),mammalian target of rapamycin(mTOR)mRNA and protein expression. RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of human periodontal ligament cells in the H2O2 group decreased.Compared with the H2O2 group,the proliferation activity of human periodontal ligament cells increased in the H2O2+Rg1 group.(2)Compared with the control group,the expression of reactive oxygen species and heme oxygenase-1 increased in the H2O2 group.Compared with the H2O2 group,the expression of reactive oxygen species and heme oxygenase-1 decreased in the H2O2+Rg1 group.(3)Compared with the control group,the expression of Caspase-3,Bax,Beclin-1 and LC3 increased,while the expression of Bcl-2 and P62 decreased in human periodontal ligament cells of the H2O2 group.Compared with the H2O2 group,the expressions of Caspase-3,Bax,Beclin-1 and LC3 decreased,and the expressions of Bcl-2 and P62 increased in the H2O2+Rg1 group.(4)Compared with the control group,the expressions of PI3K,AKT and mTOR decreased in human periodontal ligament cells of the H2O2 group.Compared with the H2O2 group,the expressions of PI3K,AKT and mTOR increased in human periodontal ligament cells of the H2O2+Rg1 group.(5)These results suggest that ginsenoside Rg1 can inhibit H2O2-induced apoptosis and autophagy in human periodontal ligament cells by reducing the content of reactive oxygen species and down-regulating the related factor expression of the PI3K/AKT/mTOR pathway.
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BACKGROUND:Rotator cuff muscle degeneration(muscle atrophy,fibrosis and fatty infiltration)is a common condition after rotator cuff tears,which seriously affects shoulder function and surgical outcomes.Ginsenoside Rg1 has biological effects such as anti-oxidation,anti-apoptosis and lipid-lowering.However,the effect of ginsenoside Rg1 on muscle degeneration after rotator cuff tear has not been reported. OBJECTIVE:To investigate the effect of ginsenoside Rg1 on muscle degeneration after massive rotator cuff tear in mice. METHODS:Sixty C57BL/6J mice were randomly divided into sham group,model group,ginsenoside Rg1 low dose group and ginsenoside Rg1 high dose group,with 15 mice in each group.The skin of the right shoulder of mice in the sham group was cut and sutured.Massive rotator cuff tear mouse models of the right shoulder were established in the other three groups.Supraspinatus tendon and suprascapular nerve compression were administrated.Mice in the sham and model groups were intraperitoneally injected with 0.5 mL of saline after operation,while those in the ginsenoside Rg1 low and high dose groups were intraperitoneally injected with ginsenoside Rg1 30 and 60 mg/kg respectively,once a day,for 6 weeks.Mice were assessed for limb function by gait analysis the day after the last injection.After euthanasia,the supraspinatus muscle on the operated side was taken to measure the muscle atrophy rate and muscle contractility.Muscle tissue was stained with oil red O and Masson.RT-PCR was used to detect the expression of atrophy,fibrosis,and fatty infiltration related genes. RESULTS AND CONCLUSION:Compared with the model group,low-and high-dose ginsenoside Rg1 significantly increased paw print area and step length(P<0.05).Compared with the model group,low-and high-dose ginsenoside Rg1 significantly increased myofiber cross-sectional area and supraspinatus contractility(P<0.05),and significantly decreased wet muscle mass reduction ratio,fatty infiltration area ratio,and collagen fiber area ratio(P<0.05).Compared with the model group,low-and high-dose ginsenoside Rg1 significantly decreased the expression of atrophy,fibrosis,and fatty infiltration related genes(P<0.05).There was no significant difference in paw print area,supraspinatus muscle contractility,and myofiber cross-sectional area between ginsenoside Rg1 low and high dose groups(P>0.05),and all other indexes were better in the ginsenoside Rg1 high dose group than in the ginsenoside Rg1 low dose group(P<0.05).To conclude,ginsenoside Rg1 could significantly reduce muscle atrophy,fibrosis and fatty infiltration following massive rotator cuff tear in mice,which is beneficial to improve muscle strength and limb function.
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ObjectiveTo investigate the influence of concentration ratio(CR) between the internal reference and target components on the quantitative accuracy of quantitative analysis of multi-components by single marker(QAMS) by taking ginsenosides as an example. MethodUltra performance liquid chromatography(UPLC) was employed, the contents of nine components in Ginseng Radix et Rhizoma(ginsenosides Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2, Rb3, Rd) were determined by external standard method(ES). Using ginsenoside Rg1 as the internal reference, the contents of the remaining 8 ginsenosides were determined by QAMS, and the quantitative results were compared with those of ES to evaluate the quantitative accuracy of the established QAMS. According to the contents of these 9 ginsenosides, the simulated sample solutions with different CRs of ginsenoside Rg1 to ginsenosides Rf, Rb2, Rd were prepared with the reference substance(CR=100∶1, 10∶1, 5∶1, 2∶1, 1∶1, 0.5∶1, 0.25∶1), in order to validate the effect of the CRs between the internal reference and other components on the quantitative accuracy of the QAMS. ResultThe results of ginsenosides Re, Rf, Rb1, Rc, Rb2 calculated by the two methods were the same with the relative standard deviation(RSD)<3%, however, the results of ginsenosides Rh1, Rb3 and Rd calculated by the two methods were different with RSDs of 7.06%-9.61%. According to the result of the simulated sample solution, the difference between the calculated results of ES and QAMS was large when the CR between the internal reference(ginsenoside Rg1) and other components was 5 or 10 or even higher. ConclusionThe quantitative error of QAMS will increase when the CR between the quantitative component and the internal reference is too large, so it is suggested that when establishing the QAMS, the components with close concentration to the internal reference should be selected for quantification.
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ObjectiveTo analyze the quantity-quality transfer of standard decoction of Ginseng Radix et Rhizoma(GRR) decoction pieces produced by fresh and traditional cutting, and to provide reference for quality control and application development of the decoction pieces produced by fresh cutting. MethodTen batches of representative GRR decoction pieces produced by fresh and traditional cutting and their standard decoctions were prepared by standard process, and high performance liquid chromatography(HPLC) fingerprint of the standard decoction was established and performed on an Agilent EC-C18 column(4.6 mm×150 mm, 2.7 μm) with acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) as the mobile phase for gradient elution(0-23 min, 18%-21%A; 23-35 min, 21%-28%A; 35-80 min, 28%-32%A), and the detection wavelength was 203 nm. Then similarity evaluation, principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) of fingerprint of the standard decoction were performed to screen the differential components with variable importance in the projection(VIP) value>1. Quantitative analysis was carried out on the screened known differential components, and combined with the indicators of the dry extract rate and the transfer rate, to explore the differences in the quantity-quality transfer between the standard decoction of GRR decoction pieces produced by fresh and traditional cutting. ResultThe fingerprint similarity of the standard decoction of GRR decoction pieces produced by fresh and traditional cutting was more than 0.950, and 18 common peaks were identified, including 9 identified common peaks. The results of PCA and PLS-DA showed that there were some differences in the contents of index components between the two standard decoctions. The contents of ginsenoside Rg1, Re and Ro in GRR decoction pieces produced by fresh cutting were higher than those in traditional decoction pieces, while the contents of ginsenoside Rb1, Rc , Rb2 and Rd were lower than those in traditional decoction pieces. The contents of ginsenoside Rg1, Re, Rb1 and Ro in the standard decoction of GRR decoction pieces produced by fresh cutting were higher than those in the standard decoction of traditional decoction pieces, while the contents of ginsenoside Rc , Rb2 and Rd were comparable between the two standard decoctions. Compared with the standard decoction of the traditional decoction pieces, the average transfer rates of ginsenoside Rg1, Rb1, Rc, Rb2 and dry extract rate of the standard decoction of GRR decoction pieces produced by fresh cutting were significantly increased(P<0.05), and the average transfer rate of ginsenoside Re and Rd also increased, but the difference was not statistically significant. ConclusionThe dry extract rate, content and transfer rate of index components of standard decoction of GRR decoction pieces produced by fresh cutting are better than those of the standard decoction of traditional decoction pieces, which can provides data support for the subsequent clinical application of fresh cutting products.
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ObjectiveTo analyze the quantity-quality transfer of standard decoction of Ginseng Radix et Rhizoma(GRR) decoction pieces produced by fresh and traditional cutting, and to provide reference for quality control and application development of the decoction pieces produced by fresh cutting. MethodTen batches of representative GRR decoction pieces produced by fresh and traditional cutting and their standard decoctions were prepared by standard process, and high performance liquid chromatography(HPLC) fingerprint of the standard decoction was established and performed on an Agilent EC-C18 column(4.6 mm×150 mm, 2.7 μm) with acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) as the mobile phase for gradient elution(0-23 min, 18%-21%A; 23-35 min, 21%-28%A; 35-80 min, 28%-32%A), and the detection wavelength was 203 nm. Then similarity evaluation, principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) of fingerprint of the standard decoction were performed to screen the differential components with variable importance in the projection(VIP) value>1. Quantitative analysis was carried out on the screened known differential components, and combined with the indicators of the dry extract rate and the transfer rate, to explore the differences in the quantity-quality transfer between the standard decoction of GRR decoction pieces produced by fresh and traditional cutting. ResultThe fingerprint similarity of the standard decoction of GRR decoction pieces produced by fresh and traditional cutting was more than 0.950, and 18 common peaks were identified, including 9 identified common peaks. The results of PCA and PLS-DA showed that there were some differences in the contents of index components between the two standard decoctions. The contents of ginsenoside Rg1, Re and Ro in GRR decoction pieces produced by fresh cutting were higher than those in traditional decoction pieces, while the contents of ginsenoside Rb1, Rc , Rb2 and Rd were lower than those in traditional decoction pieces. The contents of ginsenoside Rg1, Re, Rb1 and Ro in the standard decoction of GRR decoction pieces produced by fresh cutting were higher than those in the standard decoction of traditional decoction pieces, while the contents of ginsenoside Rc , Rb2 and Rd were comparable between the two standard decoctions. Compared with the standard decoction of the traditional decoction pieces, the average transfer rates of ginsenoside Rg1, Rb1, Rc, Rb2 and dry extract rate of the standard decoction of GRR decoction pieces produced by fresh cutting were significantly increased(P<0.05), and the average transfer rate of ginsenoside Re and Rd also increased, but the difference was not statistically significant. ConclusionThe dry extract rate, content and transfer rate of index components of standard decoction of GRR decoction pieces produced by fresh cutting are better than those of the standard decoction of traditional decoction pieces, which can provides data support for the subsequent clinical application of fresh cutting products.
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OBJECTIVE@#To obtain detailed understanding on the gene regulation of natural compounds in altering prognosis of head and neck squamous cell carcinomas (HNSC).@*METHODS@#Gene expression data of HNSC samples and peripheral blood mononuclear cells (PBMCs) of HNSC patients were collected from Gene Expression Omnibus (GEO). Differential gene expression analysis of GEO datasets were achieved by the GEO2R tool. Common differentially expressed gerres (DEGs) were screened by comparing DEGs of HNSC with those of PBMCs. The combination was further analyzed for regulating pathways and biological processes that were affected.@*RESULTS@#Totally 110 DEGs were retrieved and identified to be involved in biological processes related to tumor regulation. Then 102 natural compounds were screened for a combination such that the expression of all 110 commonly DEGs was altered. A combination of salidroside, ginsenoside Rd, oridonin, britanin, and scutellarein was chosen. A multifaceted, multi-dimensional tumor regression was showed by altering autophagy, apoptosis, inhibiting cell proliferation, angiogenesis, metastasis and inflammatory cytokines production.@*CONCLUSIONS@#This study has helped develop a unique combination of natural compounds that will markedly reduce the propensity of development of drug resistance in tumors and immune evasion by tumors. The result is crucial to developing a combinatorial natural therapeutic cocktail with accentuated immunotherapeutic potential.
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Humanos , Leucocitos Mononucleares , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Carcinoma de Células Escamosas de Cabeza y Cuello/tratamiento farmacológico , Inmunoterapia , PronósticoRESUMEN
Ginsenoside Rg1 is one of the most important saponins in ginseng. It has a wide range of pharmacological activities. It is considered to be a powerful neuroprotective agent. It has neuroprotective effects such as anti-neuroinflammation, anti-oxidative stress, anti-neuronal apoptosis, and enhancing memory. Rg1 shows a good application prospect in the prevention and treatment of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, stroke, and mental diseases such as depression. This paper reviews the research on the neuroprotective mechanism of Rg1 at home and abroad in recent years, in order to provide new research ideas for the clinical treatment of nervous system diseases.
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@#To investigate whether rare ginsenosides could alleviate idiopathic pulmonary fibrosis (IPF), C57BL/6 mice were randomly divided into control group, bleomycin (BLM)-induced IPF group, rare ginsenoside Rk1 group, rare ginsenoside Rk3 group, rare ginsenoside Rh4 group and rare ginsenoside Rg5 group.All mice except those in the control group were given bleomycin injection.The IPF model was established by BLM for 28 days.The treatment group was given ginsenoside intragastrically at the same time.After the experiment, the lung tissues of mice were collected and the pathological changes of the mice lungs were observed.The content of hydroxyproline (HYP) in mouse lung tissue was measured.The expression of IPF-related genes in mouse lung tissues was detected.In in vitro experiments, Medical Research Council cell strain-5 (MRC-5) was used to induce IPF cell model using transforming growth factor-β1 (10 ng/mL).The effects of four saponins on the expression of IPF-related genes were analyzed by MTT assay, HYP content determination and RT-qPCR.All four rare ginsenosides could effectively alleviate the pathological process such as alveolar structure destruction caused by IPF, reduce the content of HYP, and down-regulate the expression of IPF-related genes, indicating that rare ginsenosides can effectively alleviate IPF.
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The aim of this study was to investigate the therapeutic effects and potential mechanism of c(RGDyK) peptide modified mesenchymal stem cell exosomes loaded with ginsenoside Rg1 (G-Rg1) on ischemic stroke. Thread-tying method was used to establish SD rats transient middle cerebral occlusion model (tMCAO). The model rats were randomly divided into tMCAO group, Exo group, free G-Rg1 group, Exo-Rg1 group and cRGD-Exo-Rg1 group, and sham group was used as control. The infarct volume was measured by 2, 3, 5-triphenyltetrachloride (TTC) staining, the changes of neuron and endothelium were observed by immunofluorescence, and the expression of related proteins was detected by Western blotting. The results showed that cRGD-Exo-Rg1 up-regulated the expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factors (HIF-1α) by activating PI3K/AKT pathway, thus promoting angiogenesis and neurogenesis, effectively reducing the volume of cerebral infarction and improving neural function. In addition, the delivery of cRGD-Exo-Rg1 to ischemic brain tissue up-regulated the expression of occludin and claudin-5, and reduced the injury of blood-brain barrier. Taken together, cRGD-Exo-Rg1 was effective in the treatment of ischemic stroke by promoting angiogenesis and neurogenesis, which provided experimental evidence for the potential clinical benefits of other neuroprotective therapies.
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Ratas , Animales , Accidente Cerebrovascular Isquémico/tratamiento farmacológico , Ratas Sprague-Dawley , Fosfatidilinositol 3-Quinasas , Factor A de Crecimiento Endotelial Vascular/metabolismo , Exosomas/metabolismo , Ginsenósidos/uso terapéuticoRESUMEN
Ginsenoside Compound K (CK) has anti-cancer and anti-inflammatory pharmacological activities. It has not been isolated from natural ginseng and is mainly prepared by deglycosylation of protopanaxadiol. Compared with the traditional physicochemical preparation methods, the preparation of CK by hydrolysis with protopanaxadiol-type (PPD-type) ginsenoside hydrolases has the advantages of high specificity, environmental-friendliness, high efficiency and high stability. In this review, the PPD-type ginsenoside hydrolases were classified into three categories based on the differences in the glycosyl-linked carbon atoms of the hydrolase action. It was found that most of the hydrolases that could prepare CK were PPD-type ginsenoside hydrolase type Ⅲ. In addition, the applications of hydrolases in the preparation of CK were summarized and evaluated to facilitate large-scale preparation of CK and its development in the food and pharmaceutical industries.
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Ginsenósidos/farmacología , Hidrolasas , Sapogeninas/químicaRESUMEN
Via network pharmacology, molecular docking, and cellular experiment, this study explored and validated the potential molecular mechanism of ginsenoside Rg_1(Rg_1) against radiation enteritis. Targets of Rg_1 and radiation enteritis were retrieved from BATMAN-TCM, SwissTargetPrediction, and GeneCards. Cytoscape 3.7.2 and STRING were employed for the construction of protein-protein interaction(PPI) network for the common targets, and screening of core targets. DAVID was used for Gene Ontology(GO) term and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment to predict the possible mechanism, followed by molecular docking of Rg_1 with core targets and cellular experiment. For the cellular experiment, ~(60)Co-γ irradiation was performed for mo-deling of IEC-6 cells, which were then treated with Rg_1, protein kinase B(AKT) inhibitor LY294002, and other drugs to verify the effect and mechanism of Rg_1. The results showed that 29 potential targets of Rg_1, 4 941 disease targets, and 25 common targets were screened out. According to the PPI network, the core targets were AKT1, vascular endothelial growth factor A(VEGFA), heat shock protein 90 alpha family class A member 1(HSP90AA1), Bcl-2-like protein 1(BCL2L1), estrogen receptor 1(ESR1), etc. The common targets were mainly involved in the GO terms such as positive regulation of RNA polymerase Ⅱ promoter transcription, signal transduction, positive regulation of cell proliferation, and other biological processes. The top 10 KEGG pathways included phosphoinositide 3-kinase(PI3K)/AKT pathway, RAS pathway, mitogen-activated protein kinase(MAPK) pathway, Ras-proximate-1(RAP1) pathway, and calcium pathway, etc. Molecular docking showed that Rg_1 had high binding affinity to AKT1, VEGFA, HSP90AA1, and other core targets. Cellular experiment indicated that Rg_1 can effectively improve cell viability and survival, decrease apoptosis after irradiation, promote the expression of AKT1 and B-cell lymphoma-extra large(BCL-XL), and inhibit the expression of the pro-apoptotic protein Bcl-2-associated X protein(BAX). In conclusion, through network pharmacology, molecular docking, and cellular experiment, this study verified the ability of Rg_1 to reduce radiation enteritis injury. The mechanism was that it regulated PI3K/AKT pathway, thereby suppressing apoptosis.
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Humanos , Proteínas Proto-Oncogénicas c-akt/genética , Farmacología en Red , Ginsenósidos/farmacología , Fosfatidilinositol 3-Quinasas/genética , Factor A de Crecimiento Endotelial Vascular , Simulación del Acoplamiento Molecular , Traumatismos por Radiación , Medicamentos Herbarios Chinos/farmacologíaRESUMEN
This study aims to explore the neuroprotective mechanism of ginsenoside Re(GS-Re) on drosophila model of Parkinson's disease(PD) induced by rotenone(Rot). To be specific, Rot was used to induce PD in drosophilas. Then the drosophilas were grouped and respectively treated(GS-Re: 0.1, 0.4, 1.6 mmol·L~(-1); L-dopa: 80 μmol·L~(-1)). Life span and crawling ability of drosophilas were determined. The brain antioxidant activity [content of catalase(CAT), malondialdehyde(MDA), reactive oxygen species(ROS), superoxide dismutase(SOD)], dopamine(DA) content, and mitochondrial function [content of adenosine triphosphate(ATP), NADH:ubiquinone oxidoreductase subunit B8(NDUFB8) Ⅰ activity, succinate dehydrogenase complex, subunit B(SDHB) Ⅱ activity] were detected by enzyme-linked immunosorbent assay(ELISA). The number of DA neurons in the brains of drosophilas was measured with the immunofluorescence method. The levels of NDUFB8 Ⅰ, SDHB Ⅱ, cytochrome C(Cyt C), nuclear factor-E2-related factor 2(Nrf2), heme oxygenase-1(HO-1), B-cell lymphoma/leukemia 2(Bcl-2)/Bcl-2-assaciated X protein(Bax), and cleaved caspase-3/caspase-3 in the brain were detected by Western blot. The results showed that model group [475 μmol·L~(-1) Rot(IC_(50))] demonstrated significantly low survival rate, obvious dyskinesia, small number of neurons and low DA content in the brain, high ROS level and MDA content, low content of SOD and CAT, significantly low ATP content, NDUFB8 Ⅰ activity, and SDHB Ⅱ activity, significantly low expression of NDUFB8 Ⅰ, SDHB Ⅱ, and Bcl-2/Bax, large amount of Cyt C released from mitochondria to cytoplasm, low nuclear transfer of Nrf2, and significantly high expression of cleaved caspase-3/caspase-3 compared with the control group. GS-Re(0.1, 0.4, and 1.6 mmol·L~(-1)) significantly improved the survival rate of PD drosophilas, alleviated the dyskinesia, increased DA content, reduced the loss of DA neurons, ROS level, and MDA content in brain, improved content of SOD and CAT and antioxidant activity in brain, maintained mitochondrial homeostasis(significantly increased ATP content and activity of NDUFB8 Ⅰ and SDHB Ⅱ, significantly up-regulated expression of NDUFB8 Ⅰ, SDHB Ⅱ, and Bcl-2/Bax), significantly reduced the expression of Cyt C, increased the nuclear transfer of Nrf2, and down-regulated the expression of cleaved caspase-3/caspase-3. In conclusion, GS-Re can significantly relieve the Rot-induced cerebral neurotoxicity in drosophilas. The mechanism may be that GS-Re activates Keap1-Nrf2-ARE signaling pathway by maintaining mitochondrial homeostasis, improves antioxidant capacity of brain neurons, then inhibits mitochondria-mediated caspase-3 signaling pathway, and the apoptosis of neuronal cells, thereby exerting the neuroprotective effect.
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Animales , Especies Reactivas de Oxígeno/metabolismo , Antioxidantes/farmacología , Estrés Oxidativo , Factor 2 Relacionado con NF-E2/metabolismo , Caspasa 3/metabolismo , Enfermedad de Parkinson/genética , Proteína X Asociada a bcl-2/metabolismo , Fármacos Neuroprotectores/farmacología , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Drosophila/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Apoptosis , Superóxido Dismutasa/metabolismo , Adenosina Trifosfato/farmacologíaRESUMEN
Baby Boom(BBM) gene is a key regulatory factor in embryonic development and regeneration, cell proliferation, callus growth, and differentiation promotion. Since the genetic transformation system of Panax quinquefolius is unstable with low efficiency and long period, this study attempted to transfer BBM gene of Zea mays to P. quinquefolius callus by gene gunship to investigate its effect on the callus growth and ginsenoside content, laying a foundation for establishing efficient genetic transformation system of P. quinquefolius. Four transgenic callus of P. quinquefolius with different transformation events were obtained by screening for glufosinate ammonium resistance and molecular identification by PCR. The growth state and growth rate of wild-type and transgenic callus were compared in the same growth period. The content of ginsenoside in transgenic callus was determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS). The results showed that transgenic callus growth rate was significantly higher than that of wild-type callus. In addition, the content of ginsenoside Rb_1, Rg_1, Ro, and Re was significantly higher than that in wild-type callus. The paper preliminarily proved the function of BBM gene in promoting growth rate and increasing ginsenoside content, which provided a scientific basis to establish a stable and efficient genetic transformation system for Panax plants in the future.
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Femenino , Embarazo , Humanos , Ginsenósidos , Panax/genética , Cromatografía Liquida , Espectrometría de Masas en Tándem , Proliferación CelularRESUMEN
Ginsenoside Rg_3, an active component of traditional Chinese medicine(TCM), was used as the substitute for cholesterol as the membrane material to prepare the ginsenoside Rg_3-based liposomes loaded with dihydroartemisinin and paclitaxel. The effect of the prepared drug-loading liposomes on triple-negative breast cancer in vitro was evaluated. Liposomes were prepared with the thin film hydration method, and the preparation process was optimized by single factor experiments. The physicochemical properties(e.g., particle size, Zeta potential, and stability) of the liposomes were characterized. The release behaviors of drugs in different media(pH 5.0 and pH 7.4) were evaluated. The antitumor activities of the liposomes were determined by CCK-8 on MDA-MB-231 and 4T1 cells. The cell scratch test was carried out to evaluate the effect of the liposomes on the migration of MDA-MB-231 and 4T1 cells. Further, the targeting ability of liposomes and the mechanism of lysosome escape were investigated. Finally, H9c2 cells were used to evaluate the potential cardiotoxicity of the preparation. The liposomes prepared were spheroid, with uniform particle size distribution, the ave-rage particle size of(107.81±0.01) nm, and the Zeta potential of(2.78±0.66) mV. The encapsulation efficiency of dihydroartemisinin and paclitaxel was 57.76%±1.38% and 99.66%±0.07%, respectively, and the total drug loading was 4.46%±0.71%. The accumulated release of dihydroartemisinin and paclitaxel from the liposomes at pH 5.0 was better than that at pH 7.4, and the liposomes could be stored at low temperature for seven days with good stability. Twenty-four hours after administration, the inhibition rates of the ginsenoside Rg_3-based liposomes loaded with dihydroartemisinin(70 μmol·L~(-1)) and paclitaxel on MDA-MB-231 and 4T1 cells were higher than those of the positive control(adriamycin) and free drugs(P<0.01). Compared with free drugs, liposomes inhibited the migration of MDA-MB-231 and 4T1 cells(P<0.05). Liposomes demonstrated active targeting and lysosome escape. In particular, liposomes showed lower toxicity to H9c2 cells than free drugs(P<0.05), which indicated that the preparation had the potential to reduce cardiotoxicity. The findings prove that ginsenoside Rg_3 characterized by the combination of drug and excipient is an ideal substitute for lipids in liposomes and promoted the development of innovative TCM drugs for treating cancer.
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Humanos , Paclitaxel/farmacología , Liposomas/química , Ginsenósidos/uso terapéutico , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Cardiotoxicidad/tratamiento farmacológico , Línea Celular TumoralRESUMEN
Through the non-targeted metabolomics study of endogenous substances in the liver and serum of hyperlipidemia rats, the biomarkers related to abnormal lipid metabolism in hyperlipidemia rats were found, and the target of ginsenoside Rb_1 in improving hyperlipidemia was explored and its mechanism was elucidated. The content of serum biochemical indexes of rats in each group was detected by the automatic biochemical analyzer. The metabolite profiles of liver tissues and serum of rats were analyzed by HPLC-MS. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to compare and analyze the metabolic data in the normal group, the hyperlipidemia group, and the ginsenoside Rb_1 group, and screen potential biomar-kers. The related metabolic pathways were further constructed by KEGG database analysis. The results showed that hyperlipemia induced dyslipidemia in rats, which was alleviated by ginsenoside Rb_1. The non-targeted metabolomics results showed that there were 297 differential metabolites in the liver tissues of hyperlipidemia rats, 294 differential metabolites in the serum samples, and 560 diffe-rential metabolites in the hyperlipidemia rats treated by ginsenoside Rb_1. Perillic acid and N-ornithyl-L-taurine were common metabolites in the liver and serum samples, which could be used as potential biomarkers for ginsenoside Rb_1 in the improvement of hyperlipidemia. As revealed by pathway enrichment in the liver and serum, ginsenoside Rb_1 could participate in the metabolic pathway of choline in both the liver and serum. In addition, ginsenoside Rb_1 also participated in the ABC transporter, alanine, aspartic acid, and glutamate metabolism, protein digestion and absorption, β-alanine metabolism, taurine and hypotaurine metabolism, caffeine metabolism, valine, leucine, and isoleucine biosynthesis, arachidonic acid metabolism, and methionine and cysteine metabolism to improve dyslipidemia in rats.
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Ratas , Animales , Hiperlipidemias/tratamiento farmacológico , Metaboloma , Ginsenósidos/metabolismo , Metabolismo de los Lípidos , Metabolómica/métodos , Hígado/metabolismo , Biomarcadores , TaurinaRESUMEN
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease affecting both upper and lower motor neurons in the brain and spinal cord. One important aspect of ALS pathogenesis is superoxide dismutase 1 (SOD1) mutant-mediated mitochondrial toxicity, leading to apoptosis in neurons. This study aimed to evaluate the neural protective synergistic effects of ginsenosides Rg1 (G-Rg1) and conditioned medium (CM) on a mutational SOD1 cell model, and to explore the underlying mechanisms. We found that the contents of nerve growth factor, glial cell line-derived neurotrophic factor, and brain-derived neurotrophic factor significantly increased in CM after human umbilical cord mesenchymal stem cells (hUCMSCs) were exposed to neuron differentiation reagents for seven days. CM or G-Rg1 decreased the apoptotic rate of SOD1G93A-NSC34 cells to a certain extent, but their combination brought about the least apoptosis, compared with CM or G-Rg1 alone. Further research showed that the anti-apoptotic protein Bcl-2 was upregulated in all the treatment groups. Proteins associated with mitochondrial apoptotic pathways, such as Bax, caspase 9 (Cas-9), and cytochrome c (Cyt c), were downregulated. Furthermore, CM or G-Rg1 also inhibited the activation of the nuclear factor-kappa B (NF-κB) signaling pathway by reducing the phosphorylation of p65 and IκBα. CM/G-Rg1 or their combination also reduced the apoptotic rate induced by betulinic acid (BetA), an agonist of the NF-κB signaling pathway. In summary, the combination of CM and G-Rg1 effectively reduced the apoptosis of SOD1G93A-NSC34 cells through suppressing the NF-κB/Bcl-2 signaling pathway (Fig. 1 is a graphical representation of the abstract).
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Humanos , FN-kappa B/metabolismo , Ginsenósidos/farmacología , Esclerosis Amiotrófica Lateral/genética , Medios de Cultivo Condicionados/farmacología , Superóxido Dismutasa-1 , Enfermedades Neurodegenerativas , Neuronas/metabolismo , ApoptosisRESUMEN
Ginsenoside Compound K(CK)is a kind of protopanaxadiol ginsenosides,which exerts antitumor effects on various cancers,such as lung cancer,liver cancer and breast cancer.Pharmacological studies have proved that CK induces tumor cell cycle arrest and apoptosis,as well as regulates tumor cell autophagy and inhibits tumor metastasis by regulating multiple signaling pathways(AMPK/mTOR and PI3K/Akt et cetera).However,as an intestinal metabolite of natural protopanaxadiol ginsenosides,CK cannot be directly extracted from the ginseng plant.Therefore,biotransformation from natural ginsenosides such as ginsenoside Rb1 or biosynthesis are utilized to obtain CK.Biotransformation of CK uses enzymes or microorganisms to transform different ginsenosides or their structural analogs into CK;biosynthesis of CK cultures cell factories and biosynthetic enzymes to synthesize glucose and other simple compounds into CK.In this review,we systematically summarized the research progress in biopreparation and antitumor mechanism of CK in recent years,with the aim of providing evidence for the future development of CK as a clinical anti-tumor candidate drug or an adjunctive drug.
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Objective To explore the effects of ginsenoside Rg1 on blood-brain barrier, neuroinflammation and behavioral function of traumatic brain injury (TBI) mouse model.MethodsThe experiment was divided into two parts. In the first part, 27 SPF male BALB/c mice were randomly divided into blank group, sham operation group and TBI model group, with 9 mice in each group. TBI model group was made by controlled cortical impact (CCI) after craniotomy, while sham operation group was only performed craniotomy without any treatment, and the blank group was not treated at all. The effect of modeling was evaluated after operation. In the second part, 50 male BALB/c mice were randomly divided into sham operation group, three different drug dosage groups and solvent (DMSO) control group, with 8 mice in each group. The drug treatment groups were injected with ginsenoside Rg1 at the doses of 10, 20 and 40 mg/kg respectively 6 hours after TBI model had been successfully established, while the DMSO control group was given the same amount of 1% DMSO for one week, twice a day. Modified neurological severity scores (mNSS) were performed on the 1st, 3rd, 7th and 14th day after modeling, and the blood-brain barrier leakage was detected by Western blotting on the 3rd day after modeling. On the 14th and 16th day, the elevated cross maze test and water maze test were used to detect the neurobehavioral function. On the 28th day after anesthesia and perfusion, the brains were taken out, and the neuroinflammation such as activation of microglia and astrocytes was observed by immunofluorescence staining.ResultsThe expression level of MMP-9, a marker of blood-brain barrier, decreased in ginsenoside Rg1 treatment group (P<0.01). The number of microglia (Iba-1 positive) and astrocyte (GFAP positive) cells decreased significantly (P<0.05), which indicated that neuroinflammation was inhibited, and the best effect was achieved at the dosage of 20 mg/kg (P<0.01). The mNSS of mice in ginsenoside Rg1 treatment group were significantly lower than those in DMSO control group (P < 0.01), and the proportion of times they entered the open arm was significantly higher than that in DMSO control group (P < 0.05). The time ratio in the quadrant where the water maze experimental platform was located and the times of crossing the platform were significantly higher than those in control group (P < 0.05), and the dosage of 20 mg/kg had the best effect.ConclusionThe TBI mouse model was successfully constructed and applied to the study of ginsenoside Rg1 repair of mouse traumatic brain injury. Ginsenoside Rg1 can significantly improve blood-brain barrier, alleviate neuroinflammation and improve neurobehavioral function in TBI model mice, and the effect is the most significant at the dose of 20 mg/kg.
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Hepatocellular carcinoma(HCC)is the third leading cause of cancer death worldwide.Ginsenoside Rk3,an important and rare saponin in heat-treated ginseng,is generated from Rg1 and has a smaller mo-lecular weight.However,the anti-HCC efficacy and mechanisms of ginsenoside Rk3 have not yet been characterized.Here,we investigated the mechanism by which ginsenoside Rk3,a tetracyclic triterpenoid rare ginsenoside,inhibits the growth of HCC.We first explored the possible potential targets of Rk3 through network pharmacology.Both in vitro(HepG2 and HCC-LM3 cells)and in vivo(primary liver cancer mice and HCC-LM3 subcutaneous tumor-bearing mice)studies revealed that Rk3 significantly inhibits the proliferation of HCC.Meanwhile,Rk3 blocked the cell cycle in HCC at the G1 phase and induced autophagy and apoptosis in HCC.Further proteomics and siRNA experiments showed that Rk3 regulates the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway to inhibit HCC growth,which was validated by molecular docking and surface plasmon resonance.In conclusion,we report the discovery that ginsenoside Rk3 binds to PI3K/AKT and promotes autophagy and apoptosis in HCC.Our data strongly support the translation of ginsenoside Rk3 into novel PI3K/AKT-targeting ther-apeutics for HCC treatment with low toxic side effects.
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OBJECTIVE Microgravity exerts several negative effects on the learning and memory of astro-nauts during space flight.Rg1 and Rb1,the key steroidal components of ginseng,have shown potent neuroprotec-tive effects with a high safety profile.The object of the current study is to investigate the influence of Rg1 and Rb1 on simulated microgravity-induced memory and learning dysfunction in the hindlimb suspension(HLS)rat model.METHODS The HLS rats were orally administered Rg1(30 and 60 μmol·kg-1)or Rb1(30 and 60 μmol·kg-1)for four weeks.The Morris water maze test(MWM)and reward operating conditioning reflex test(ROCR)were conducted to evaluate spatial and associative learning and memory.After the behavior tests,the serum and the prefrontal cortex(PFC)were dissected to measure the mechanism.RESULTS Rg1 and Rb1 treatment amelio-rated the cognitive deficits of HLS-exposure rats in MWM and ROCR,reduced reactive oxygen species generation and increased antioxidant enzyme activity.Rg1 and Rb1 also assisted in the recovery of mitochondrial complex Ⅰ(NADH dehydrogenase)activities and Mfn2,and decrea-sed Drp-1 expression.Furthermore,Rg1 and Rb1 reduced the ratio of Bax/Bcl-2 and the expression of cleaved-cas-pase 3,cytochrome c,increased the levels of SYN,PSD95 and activated BDNF-TrkB/PI3K-Akt pathway in the PFC.CONCLUSION Rg1 and Rb1 treatment attenuated cog-nitive deficits induced by HLS,mitigated mitochondrial dysfunction,attenuated oxidative stress,inhibited apopto-sis,and increased the synaptic plasticity,which was partly mediated by the modulation of the BDNF-TrkB/PI3K-Akt signaling.