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Objective: To analyze the molecular interaction network pathway of Shenmai Injection in the treatment of COVID-19 with coronary heart disease by using network pharmacology. Methods: Using the TCMSP and ETCM to retrieve the chemical constituents of Ginseng Radix et Rhizoma Rubra and Ophiopogonis Radix in Shenmai Injection. The target of the compound was predicted through the SwissTargetPrediction database. The target of COVID-19 with coronary heart disease was screened through the NCBI database and the GeneCards database, and the targets of compound and disease were mapped to obtain the target of the compound for treating the disease. FunRich software and DAVID database were used to perform GO function enrichment analysis and KEGG pathway enrichment analysis, and Excel software and Tableau software to draw bar charts and bubble charts for visualization. Finally, Cytoscape 3.7.1 software was used to build compound-target-pathway network. Glide was used to dock the components of Shenmai Injection with 3CL hydrolase (Mpro). Results: The results showed that ophiopogonin D', ophiopogonin D, ginsenoside Rg2, methyl ophiopogonanone A, ophiogenin-3-O-α-L-rhamnopyranosyl (1→2)-β-D-glucopyranoside, ginsenoside Rb2, ginsenoside R0, ophiopogon A, sanchinoside Rd, ophiopogonanone E, and ginsenoside Re showed higher degrees in the analysis and stronger binding with 3CL hydrolase. Those compounds were the main effective components in the treatment of COVID-19 combined with coronary heart disease, involving 77 targets such as IL6, GAPDH, ALB, TNF, MAPK1, MAPK3, TP53, EGFR, CASP3, and CXCL8. KEGG pathway enrichment analysis revealed that there were 124 (P < 0.05) signaling pathways involving HIF-1 signaling pathway, TNF signaling pathway, sphingolipid signaling pathway, Toll-like receptor signaling pathway, neurotrophin signaling pathway, VEGF signaling pathway, apoptosis, Ras signaling pathway, PI3K-Akt signaling pathway, and prolactin signaling pathway. The results of molecular docking showed that the affinity between the 17 components of Shenmai Injection and the 3CL hydrolase of SARS-CoV-2 was less than -25 kJ/mol. Conclusion: Shenmai Injection can achieve simultaneous intervention of COVID-19 and coronary heart disease by inhibiting cytokine storms, maintaining cardiac function homeostasis, regulating immunity, and antivirals. It presents the network regulation mechanism of mutual influence and complex correlation. This study can provide a scientific basis for the treatment of Shenmai Injection in critically ill patients with COVID-19.
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Yiqi Fumai Lyophilized Injection is a modern preparation composed of Panax ginseng, Ophiopogon japonicas, and Schisandra chinensis. Clinically, it is mainly used for the treatment of cardiovascular diseases such as angina pectoris of coronary heart disease and chronic cardiac insufficiency. According to the concept of quality markers (Q-markers), the Q-markers of Yiqi Fumai Lyophilized Injection were predicted and analyzed from the aspects of chemical components, drug effect, network pharmacology, pharmacokinetics, and drug property. Based on the above studies, 13 components of ginsenosides Rb1, Rg1, Rf, Rh1, Rc, Rb2, Ro, Rg3, ophiopojaponin C, phiogenin-3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside, pennogenin-3-O-α-L-rhamnopyranosyl-(1→2)- β-D-xylopyranosyl-(1→4)-β-D-glucopyranoside, fructose, and schisandrin A were identified as Q-markers. According to the Q-markers, we established quality system for process control. The study of Yiqi Fumai Lyophilized Injection based on Q-markers can provide new research ideas for quality assessment of traditional Chinese materia medica injection.
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Objective: On the basis of simultaneous determination of seven saponins in flower buds of Panax ginseng, a method of quantitative analysis of multi-components by single marker (QAMS) for the determination of seven saponins was established, and the feasibility of the method was verified. Methods: Using HPLC-UV, ten batches of dried P. ginseng flowers were used as the research object. Ginsenoside Re was used as internal reference to determine the relative correction factor of ginsenoside Rg1, Rg2, Rb1, Rc, Rb2 and Rd. The content of each component was measured by the traditional external standard method, and the difference between the calculated value and the measured value was compared to verify the feasibility and accuracy of the external standard method. Results: The relative correction factors of six ginsenoside Rg1, Rg2, Rb1, Rc1, Rb2, and Rd in P. ginseng flower were 1.07, 1.05, 0.81, 0.80, 0.64, and 0.84, respectively. The relative correction factors of six ginsenosides were reproducible in the 10 batches, the determiation of QAMS were not significantly different from those measured by the external standard method. Conclusion: In the case of shortage of ginsenoside reference substance, a method of QAMS can be used, the content of ginsenoside Rg1, Rg2, Rb1, Rb1, Rb2, and Rd in flower buds of P. ginseng can be determined by relative calibration factor.
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Objective: This study aimed to simultaneously determine six composition of Panax ginseng by quantitative analysis of multi-components with a single-marker (QAMS) in different paris. Methods: Phenomenex Luna C18 (250 mm × 4.6 mm, 5 μm) was used with mobile phase consisting of acetonitrile-0.1% phosphoric acid for gradient elution at a flow rate of 1.0 mL/min, The column temperature was 25 ℃ and the detection wavelength was 203 nm. Ginsenoside Rb1 was used as reference to establish its relative correction factor of Rg1, Re, Rc, Rb2, Rd, The contents of six components were determined by both external standard method and QAMS. and t test was used to evaluate the feasibility and applicability of QAMS. Results: In a certain linear range, the relative correction factor (RCF) was good, No significant differences were observed between the quantitative results of the two methods. Conclusion: It is feasible and suitable to evaluate the quality of Panax ginseng. It can provide a useful reference to quality control of multi-indexed components in Chinese herbs and traditional Chinese preparations.
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Objective: To evaluate the quality of Maiweishen, a simple and accurate HPLC method for determining the contents of 20 active constituents from Maiweishen was established. Methods: The chromatographic separation was achieved on a C18 column (150 mm × 4.6 mm, 5 μm) using a mobile phase made up of acetonitrile and water at a flow rate of 1.0 mL/min. The detection wavelength and column temperature were set as 203 nm and 35 ℃, respectively. Results: Sixteen ginsenosides (Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, F1, Rd, F2, Rg3, protopanaxatriol, compounds K, Rh2, and protopanaxadiol), three kinds of lignan in Schisandra chinensis (schizandrol A, schizandrin A, B), and ophiopogonin D were separated at baseline with good linearity (r ≥ 0.999 6). The recovery rates were 96%-102% (RSD < 2%). Conclusion: The method is simple, fast, accurate, and could be applied to the quality control of Maiweishen.
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Objective To explore the effect of ecological factors and the expression of key enzyme genes on the synthesis and accumulation of ginsenosides. Methods Cultivated four-year-old ginseng were used as test materials, the expression of key enzyme genes (HMGR, FPS, SS, SE, DS, β-AS, CYP82D47, CYP716A47) in the biosynthesis of ginsenosides of roots in different growth periods was determined by real-time quantitative PCR, determination of the content of eight ginsenosides (ginsenoside Rg1, Re, Rf, Rb1, Rb2, Rb3, Rc, Rd) in roots by HPLC, the meteorological data were collected by a small weather station, correlation analysis was performed with SPSS. Results The expression of key enzyme genes in the period of flowering to fruit ripening was higher than the root growing after fruit period and the withering period, the expression of key enzyme genes involved in the synthesis of ginsenosides was influenced by each other, and the expression of key enzyme genes in ginseng roots showed a positive correlation with the accumulation of ginsenosides; The contents of ginsenoside Rg1, Rb1, Re, and Rc were higher in the roots of ginseng, eight kinds of monomer ginsenosides content dynamic changes trend is different; Temperature, photosynthetically active radiation, soil water potential are important ecological factors for ginsenosides synthesis in roots, temperature was significantly negatively correlated with ginsenoside Rb1 and Rd (P < 0.05), PAR can significantly promote the formation of ginsenoside Rg1 (P < 0.05), soil water potential was significantly negatively correlated with ginsenoside Rb1 (P < 0.05); Grey correlation analysis results showed that the major ecological factors that influenced ginsenosides content in ginseng roots were temperature, PAR and relative humidity, the grey correlation between the expression of the key enzyme genes with the content of ginsenosides is less than ecological factors with the content of ginsenosides, under the guidance of ecological factors, the expression of the key enzyme genes regulate the synthesis and accumulation of ginsenosides. Conclusion The dynamic changes of the expression of key enzyme genes and the content of ginsenosides in ginseng were determined, it provides a theoretical basis for elucidating the physiological and ecological mechanism of ginsenoside synthesis and the quality control of Radix Ginseng.
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AIM To prepare the transdermal drug delivery system of carbon nanotubes graft ginsenosides Rb1 and Rb2 nanoemulsion.METHODS After 0.5％ carbon nanotubes' dispersion into the prepared nanoemulsion,the combination state was observed under transmission electron microscope.The skin irritation and transdermal mechanism of this drug delivery system were investigated.Then the UVA-damaged mouse skin model was established to evaluate the antioxidant activity.RESULTS Carbon nanotubes had strong adsorption to nanoemulsion,together with the formation of drug storage cavern in epidermis,thus increased the transdermal osmotic quantities of ginsenosides Rb1 and Rb2.The SOD activity in mouse skin tissue in the drug delivery system group was higher than that in the model group and the nanoemulsion group (P ＜0.01),and the MDA content was significantly decreased (P ＜ 0.01).CONCLUSION The transdermal drug delivery system of carbon nanotubes graft ginsenosides Rb1 and Rb2 nanoemulsion can make drugs play an antioxidant role because of its less irritation to skin and better skin permeability.
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A rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometric (RRLC-Q-TOF-MS) method was established and optimized for the analysis of pharmacokinetic behavior of ginsenoside Rb2 in rats by intravenous injection administration.The metabolism of ginsenosides Rb2 in vivo rat was also explored.In the experiment,Agilent SB C18 column was selected for the sample separation with 0.1％ aqueous formic acid solution as mobile phase (A) and acetonitrile as mobile phase (B) at a flow rate of 0.2 mL/min,and the injection volume was set to 5 μL.Q-TOF-MS was carried out in electron pray ionization (ESI) negative ion mode.The limit of quantification (LOQ,S/N =10) and limit of detection (LOD,S/N=3) were 0.10 and 0.08 μg/mL,respectively,and the linear range was 0.1-1.26 μg/mL.The experiment results showed that the concentration-time profile of ginsenoside Rb2 conformed to a two-compartment pharmacokinetic model after intravenous administration for rats.The mean plasma elimination half-lives were (23.58±1.10) min (t1/2α),(1306.55±147.23) min (t1/2β) for Rb2.By analyzing the urine of rats after intravenous administration and the fecal samples after oral administration of ginsenoside Rb2,it was found that the metabolites were M6,M2 (CY),F2,and C-K.
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Objective: A high-performance liquid chromatography-tandem mass spectrometric method for the simultaneous determination of 15 ginsenoside compounds from Panacis Majoris Rhizoma (PMR) was developed. Methods: A Waters Sunfire ™ C18 column (150 mm × 4.6 mm, 5 μm) was used for the separation. The mobile phase consisted of A (H2O + 0.05% HCOOH) and B (CH3CN + 0.05% HCOOH) using a gradient elution. For the quantification of ginsenosides, the multiple reaction monitoring (MRM) mode of the mass spectrometer was applied and the declustering potential (DP), collision energy (CE), and collision cell exit potential (CXP) were optimized to perform automatic on-line MS/MS experiments during the chromatographic separation. Results: By using the optimized method, the linearity range of 15 analytes was 0.000 9 to 2 952.592 3 μg/mL with more than 0.999 determination coefficient (r) of linear regressions, the detection limits of the 15 ginsenosides ranged from 0.003 to 626.554 ng/mL, the limits of quantitation ranged from 0.075 to 1 762.150 ng/mL, the recoveries of 15 ginsenosides in the samples were 98.15%-101.12% with relative standard deviation (RSD) that ranged from 0.82% to 2.15%. Conclusion: The proposed LC-MS/MS method is accurate and reproducible in accordance with TCM guidelines, showing high sensitivity, rapidness, and recovery. This method allows the assessment of various ginsenosides in a single analytical run providing an innovative tool to control Panacis Majoris Rhizoma materials quantification.
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Objective: To select the reasonable post-processing methods after harvest of the flower of Panax ginseng (FPG) and Panax quinquefolius (FPQ). The present study evaluated the effect of different drying methods on the quality of FPG and FPQ based on the contents of 14 ginsenosides. Methods: The contents of 14 ginsenosides were quantified by HPLC including ginsenosides Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd and their corresponding malonyl-ginsenosides (m-Rg1, m-Re, m-Rb1, m-Rc, m-Rb2, m-Rb3, and m-Rd). Results: The contents of 14 ginsenosides were highest by oven drying at 40℃, followed by freeze drying, microwave drying, and infrared drying, and decreased with the increased temperature. The contents of malonyl-ginsenosides obviously decreased with reaching a high temperature (> 100℃). The decrement of malonyl-ginsenosides and the variation of corresponding ginsenosides was not equivalent, and the total content of 14 ginsenosides was also reduced. Conclusion: Oven drying at 40℃ is a suitable drying method for keeping the content of original ginsenosides, and oven drying at high temperature (> 100℃) can prompt the transformation into rare ginsenosides. Different drying methods can be selected according to different purposes in clinical application.
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Objective: To study the pharmacokinetic profiles of the nine ginsenosides from the roots of Panax ginseng in rats, such as ginsenosides Rb1, Rb2/Rb3, Rc, Rd, Re, Rf, Rg1, and Rh1. Methods: After different time points of ig administration of 200 mg/kg ginsenosides, the blood was taken from the venous plexus of fundus. The biological samples were extracted with n-butanol. Chromatographic separation was performed on a C18 column using a gradient elution program at the flow rate of 0.2 mL/min. The LC-MS system was operated using an electro-spray ionization probe in the negative ion model. After the oral administration of 200 mg/kg ginsenosides to rats, plasma was collected and analyzed under the above conditions. The pharmacokinetic parameters were calculated by non-compartment model. Results: After the oral administration of ginsenosides to rats, six ginsenosides were detected in plasma which included Rb1, Rb2/Rb3, Rc, Rd, Re, and Rg1. Among these ginsenosides, the protopanaxatriol ginsenoside Rg1 and Re were quickly eliminated. However, the pharmacokinetic behaviors of protopanaxadiol ginsenoside Rb1, Rb2/Rb3, Rc, and Rd were markedly different from those of ginsenosides Rg1 and Re in rats with the significantly longer half-life of the protopanaxadiol ginsenosides. Conclusion: The method is accurate, stable, and reliable, and can be used for profiling total ginsenosides' pharmacokinetic properties in rats.
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Objective: To establish a quantitative analysis of multi-components with a single-marker (QAMS) method for the determination of eight components in Qibai Pingfei Granule (QPG). Methods: Ginsenoside Rb1 was used as the internal reference substance. The relative correlation factors (f) of ginsenosides Rg, Re, Rf, Rc, Rb2, Rc, and astragaloside were calculated and established. The results were compared with those obtained by the external standard method in 10 batches of QPG. The results showed that the f values of ginsenosides Rg, Rf, Rc, Rb2, astragaloside, and ginsenosides Rc to ginsenoside Rb1 were 1.244, 1.075, 1.133, 1.090, 1.071, 0.967 and 1.070. Results: Ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, Rd, and astragaloside had good relations within the ranges of 0.416 0-4.992, 0.315 4-3.785 2, 0.259 6-3.115, 0.385 2-4.622, 0.222 8-2.674, 0.193 2-2.318, 0.183 5-2.202, and 0.574 6-6.895 μg, respectively. The two methods did not show the significant difference in assay results. Conclusion: So the QAMS method is feasible and credible, and could be used to determine the multiple components in QPG.
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The aim of the present study was to investigate whether ginsenoside-Rb2 (Rb2) can affect the secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla. Rb2 (3~30 microM), perfused into an adrenal vein for 90 min, inhibited ACh (5.32 mM)-evoked CA secretory response in a dose- and time-dependent fashion. Rb2 (10 microM) also time-dependently inhibited the CA secretion evoked by DMPP (100 microM, a selective neuronal nicotinic receptor agonist) and high K+ (56 mM, a direct membrane depolarizer). Rb2 itself did not affect basal CA secretion (data not shown). Also, in the presence of Rb2 (50 microg/mL), the secretory responses of CA evoked by veratridine (a selective Na+ channel activator (50 microM), Bay-K-8644 (an L-type dihydropyridine Ca2+ channel activator, 10 microM), and cyclopiazonic acid (a cytoplasmic Ca2+-ATPase inhibitor, 10 microM) were significantly reduced, respectively. Interestingly, in the simultaneous presence of Rb2 (10 microM) and L-NAME (an inhibitor of NO synthase, 30 microM), the inhibitory responses of Rb2 on ACh-evoked CA secretory response was considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of Rb2-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of Rb2 (10 microM) was greatly elevated compared to the corresponding basal released level. Collectively, these results demonstrate that Rb2 inhibits the CA secretory responses evoked by nicotinic stimulation as well as by direct membrane-depolarization from the isolated perfused rat adrenal medulla. It seems that this inhibitory effect of Rb2 is mediated by inhibiting both the influx of Ca2+ and Na+ into the adrenomedullary chromaffin cells and also by suppressing the release of Ca2+ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade.
Subject(s)
Animals , Rats , 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester , Adrenal Medulla , Calcium , Catecholamines , Chromaffin Cells , Cytoplasm , Dimethylphenylpiperazinium Iodide , Membranes , Neurons , NG-Nitroarginine Methyl Ester , Nitric Oxide Synthase , Receptors, Nicotinic , Veins , VeratridineABSTRACT
AIM: To develop a method for determining ginsenoside Rb_1,Rc,Rb_2 and Rd in Shenqi Granules (Radix Rhizoma Ginseng,Radix Astragali,Rhizoma Atractylodis macrocephalae,Poria,etc.) by microbore LC.METHODS: Microbore liquid chromatography was applied.Microsil C_(18) column( 150 mm×1.0 mm) was used with the mobile phase containing acetonitrile-water for gradient elution.The flow rate was 50 μL/min,and the detection wavelength was set at 220 nm.RESULTS : The relationships between the concentrations and the peak areas of these for components were all linear.The recoveries were 98.14% for ginsenoside Rb_1,98.04% for ginsenoside Rc,98.79% for ginsenoside Rb_2,and 97.97% for ginsenoside Rd respectively.The relative standard deviations (RSD) were 0.67%,0.99%,0.75%,and 1.46%,respectively.CONCLUSION: This method is simple,convenient and can be used for quality control.
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AIM:To develop a method for determining ginsenoside Rb_1,Rc,Rb_2 and Rd in Shenqi Granules(Radix Rhizoma Ginseng,Radix Astragali,Rhizoma Atractylodis macrocephalae,Poria,etc.)by microbore LC.METHODS:Microbore liquid chromatography was applied.Microsil C_ 18 column(150 mm?1.0 mm)was used with the mobile phase containing acetonitrile-water for gradient elution.The flow rate was 50 ?L/min,and the detection wavelength was set at 220 nm.RESULTS:The relationships between the concentrations and the peak areas of these for components were all linear.The recoveries were 98.14% for ginsenoside Rb_1,98.04% for ginsenoside Rc,98.79% for ginsenoside Rb_2,and 97.97% for ginsenoside Rd respectively.The relative standard deviations(RSD)were 0.67%,0.99%,0.75%,and 1.46%,respectively.CONCLUSION:This method is simple,convenient and can be used for quality control.