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@#Abstract: Objective To investigate the influences of notoginsenoside R1 (NGR1) on cell injury and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway of alveolar epithelial cells infected by Klebsiella pneumoniae (Kp). Methods A549 cells were grouped into five groups: control group (C group), infection group (Infect group), infection + low NGR1 group (Infect + L-NGR1 group), infection + high NGR1 group (Infect + H-NGR1 group), and infection+high NGR1+JAK2/STAT3 pathway inhibitor group (Infect+H-NGR1+SD-1029 group). Cell proliferation was measured using CCK8; ELISA kits were applied to detect the contents of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interferon γ (IFN-γ) in the culture medium; flow cytometry was applied to detect apoptosis; RT-qPCR was applied to detect the expressions of JAK2/STAT3; Western blot was applied to detect JAK2/STAT3 pathway, autophagy protein microtubule-associated protein 1 light chain 3 (LC3), autophagy-relatedgene5 (Atg5), autophagy-related gene (Atg) 6 (Beclin-1), apoptosis protein B-cell lymphoma 2 (Bcl-2), Bcl-2-accociated protein (Bax), cysteinyl aspartate specific proteinase (cleaved-caspase-3) proteins expression. Results Compared with the C group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect group were obviously decreased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Compared with Infect group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect+L-NGR1 group and Infect+H-NGR1 group were obviously increased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-Caspase-3 were obviously decreased (P<0.05). Compared with Infect+H-NGR1 group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the protein phosphorylation levels of JAK2, STAT3 in the Infect+H-NGR1+SD-1029 group were obviously decreased (P<0.05), and the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Conclusions NGR1 can activate the JAK2/STAT3 signaling pathway, promote autophagy of alveolar epithelial cells, and inhibit Kp-induced inflammatory injury and apoptosis of alveolar epithelial cells.
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The purpose of this study was to investigate the effect of notoginsenoside R_1(NGR_1) on alleviating kidney injury by regulating renal oxidative stress and the Nrf2/HO-1 signaling pathway in mice with IgA nephropathy(IgAN) and its mechanism. The mouse model of IgAN was established using a variety of techniques, including continuous bovine serum albumin(BSA) gavage, subcutaneous injections of carbon tetrachloride(CCl_4) castor oil, and tail vein injections of lipopolysaccharide(LPS). After successful modeling, mice with IgAN were randomly separated into a model group, low, medium, and high-dose NGR_1 groups, and a losartan group, and C57BL6 mice were utilized as normal controls. The model and normal groups were given phosphate buffered saline(PBS) by gavage, the NGR_1 groups were given varying dosages of NGR_1 by gavage, and the losartan group was given losartan by gavage for 4 weeks. The 24-hour urine of mice was collected after the last administration, and serum and kidney tissues of mice were taken at the end of the animal experiment. Then urine red blood cell count(URBCC), 24-hour urine protein(24 h protein), serum creatinine(Scr), and blood urea nitrogen(BUN) levels were measured. The enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of galactose-deficient IgA1(Gd-IgA1), kidney injury molecule 1(Kim-1), and neutropil gelatinase-associated lipocalin(NGAL) in the mouse serum. The assay kits were used to detect the levels of malondialdehyde(MDA) and superoxide dismutase(SOD), and immunofluorescence(IF) was used to detect the expression level of glutathione peroxidase 4(GPX4) in the mesangial region. Western blot was used to detect the protein expression of nuclear transcription factor E2 related factor 2(Nrf2)/heme oxygenase 1(HO-1) signaling pathway in the renal tissue. Hematoxylin-eosin(HE) staining was used to observe pathological alterations in the glomerulus of mice. The results revealed that, as compared with the model group, the serum Gd-IgA1 level, URBCC, 24 h protein level, renal damage markers(Kim-1 and NGAL) in the high-dose NGR_1 group decreased obviously and renal function indicators(BUN, Scr) improved significantly. The activity of SOD activity and expression level of GPX4 increased significantly in the high-dose NGR_1 group, whereas the expression level of MDA reduced and protein expression levels of Nrf2 and HO-1 increased. Simultaneously, HE staining of the renal tissue indicated that glomerular damage was greatly decreased in the high-dose NGR_1 group. In conclusion, this study has clarified that NGR_1 may alleviate the kidney injury of mice with IgAN by activating the Nrf2/HO-1 signaling pathway, improving antioxidant capacity, and reducing the level of renal oxidative stress.
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Panax notoginseng is an ancient Chinese medicinal plant that has great clinical value in regulating cardiovascular disease in China. As a single component of panax notoginosides, notoginsenoside R1 (NGR1) belongs to the panaxatriol group. Many reports have demonstrated that NGR1 exerts multiple pharmacological effects in ischemic stroke, myocardial infarction, acute renal injury, and intestinal injury. Here, we outline the available reports on the pharmacological effects of NGR1 in ischemia-reperfusion (I/R) injury. We also discuss the chemistry, composition and molecular mechanism underlying the anti-I/R injury effects of NGR1. NGR1 had significant effects on reducing cerebral infarct size and neurological deficits in cerebral I/R injury, ameliorating the impaired mitochondrial morphology in myocardial I/R injury, decreasing kidney injury molecule-1 and neutrophil gelatinase-associated lipocalin in renal I/R injury and attenuating jejunal mucosal epithelium injury in intestinal I/R injury. The various organ anti-I/R injury effects of NGR1 are mainly through the suppression of oxidative stress, apoptosis, inflammation, endoplasmic reticulum stress and promotion of angiogenesis and neurogenesis. These findings provide a reference basis for future research of NGR1 on I/R injury.
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Humans , Reperfusion Injury/prevention & control , Inflammation , China , ApoptosisABSTRACT
The present study established a quality evaluation method for ginsenoside reference substances based on quantitative nuclear magnetic resonance(qNMR) spectroscopy. ~1H-NMR spectra were collected on Bruker Avance Ⅲ 500 MHz NMR spectrometer equipped with a 5 mm BBO probe. The acquire parameters were set up as follows: pulse sequence of 30°, D_1=20 s, probe temperature= 303 K, and the scan number = 32. Dimethyl terephthalate, a high-quality ~1H-qNMR standard, was used as the internal standard and measured by the absolute quantitative method. Methyl peaks of comparatively good sensitivity were selected for quantification, and linear fitting deconvolution was adopted to improve the accuracy of integration results. The qNMR spectroscopy-based method was established and validated, which was then used for the quality evaluation of ginsenoside Rg_1, ginsenoside Re, ginsenoside Rb_1, ginsenoside Rd, and notoginsenoside R_1. The results suggested that the content of these ginsenoside reference standards obtained from the qNMR spectroscopy-based method was lower than that detected by the normalization method in HPLC provided by the manufacturers. In conclusion, the qNMR spectroscopy-based method can ensure the quality of ginsenoside reference substances and provide powerful support for the accurate quality evaluation of Chinese medicine and its preparations. The qNMR spectroscopy-based method is simple, rapid, and accurate, which can be developed for the quantitative assay of Chinese medicine standard references.
Subject(s)
Chromatography, High Pressure Liquid/methods , Ginsenosides/analysis , Magnetic Resonance Spectroscopy/methods , Proton Magnetic Resonance Spectroscopy , Reference StandardsABSTRACT
Objective:To investigate the effects of notoginsenoside R1 （NR1） on the proliferation of mice aortic smooth muscle cells （MOVAS cells） induced by angiotensinⅡ （AngⅡ） and the signal pathway of angiotensin Ⅱ type 1 receptor （AT1R） / mitogen activated protein kinases （MAPKs）. Methods:The proliferation of MOVAS cells was detected by BrdU method after AngⅡ induction. Western blot was used to detect the expression of the two main receptors of AngⅡ （AT1R and AT2R） and MAPKs pathway related proteins （ERK， p38， and JNK）. Results:（1） AngⅡ （5 μmol/L） could promote the proliferation of MOVAS cells （P<0.01）. NR1 （50 μmol/L） could inhibit the proliferation of MOVAS cells induced by AngⅡ （P<0.01）. There was no significant difference between control group and NR1 group （P>0.05）. （2） Compared with AngⅡ group， the expression of AT1R protein in AngⅡ+ NR1 group was significantly lower （P<0.05）， but there was no difference in the expression of AT2R protein （P>0.05）. （3） NR1 could significantly inhibit the phosphorylation of ERK， p38 and JNK protein after AngⅡ stimulation （P<0.01）. Conclusion:NR1 can inhibit the proliferation of MOVAS cells induced by AngⅡ， which may be related to down regulating AT1R and inhibiting the activation of MAPKs.
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BACKGROUND: Previous studies have found that panax notoginseng saponins have a certain protective effect on immunological liver injury in mice. OBJECTIVE: To explore the therapeutic effect of notoginsenoside R1 on carbon tetrachloride-induced liver fibrosis in rats. METHODS: Experimental liver fibrosis model was made by carbon tetrachloride in male Sprague-Dawley rats. Then 30 g/L notoginsenoside R1 (60 mg/kg) was given once daily for 4 and 6 weeks in the treatment group. Rats in the control and model group were given distilled water of the same volume. Histopathological observation with hematoxylin-eosin staining and Masson’s trichrome staining was used to evaluate the changes of liver structure and fibrosis degree. The expression of collage type I, α-smooth muscle actin and transforming growth factor-β1 mRNA of hepatic tissue was measured by qRT-PCR method. The experimental protocol was approved by the Animal Experiment Ethics Committee of Kunming Medical University (approval No. KMMU2018018). RESULTS AND CONCLUSION: Liver histopathology showed that notoginsenoside R1 improved the degree of liver fibrosis. The expression levels of collagen type I, α-smooth muscle actin and transforming growth factor-β1 mRNA were reduced significantly in the treatment group compared with the model group (P < 0.05). But there was no significant difference after 4 and 6 weeks of treatment with notoginsenoside R1. Overall findings indicate that notoginsenoside R1 can slow down the progression of carbon tetrachloride-induced liver fibrosis in rats to a certain extent.
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Objective: To establish a method for determination of five saponins in Panax notginseng by HPLC and comprehensively evaluate the quality of it by using grey correlation analysis. Methods: The content of notoginsenoside R1 and ginsenoside Rg1, Re, Rb1, Rd in the different origins and commercial grades of P. notginseng was simultaneously determined by HPLC, and the entire quality evaluation model was established by grey correlation analysis. Results: The established method was applied to quantify five major bioactive components in P. notginseng simultaneously with satisfactory results. Gray correlation method can distinguish the samples from genuine producing areas, qualified samples and unqualified samples, and provide reference for quality evaluation of P. notoginseng and quality evaluation of multi-index components of Chinese materia medica. Conclusion: This HPLC method was simple, accurate, stable and rapid with better separation effect, which was suitable for determination of notoginsenoside R1 and ginsenosides Rg1, Re, Rb1, Rd; The grey recognition analysis was suitable for the comprehensive quality evaluation of multi-component samples of Chinese materia medica.
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Objective To investigate whether borneol can promote the bioactive components of the combination of astragaloside IV (AST IV) and Panax notoginseng saponins (PNS) into the blood-brain barrier of rats with middle cerebral artery occlusion (MCAO)/reperfusion. Methods Using the model of MCAO/reperfusion, rats were randomly divided into sham-operation group, model group, borneol group, AST IV group, PNS group, AST IV + PNS group and borneol + AST IV + PNS group, and the content of AST IV and the bioactive components of PNS (ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1) in the cerebral cortex and the cerebellum of the affected side and the healthy side were determined by liquid chromatography-mass spectrometry (LC-MS/MS). Results AST IV, whether used alone or combined with PNS and borneol, was mainly distributed in the cerebral cortex after oral administration, especially in the affected cerebral cortex. Borneol combined with AST IV and PNS significantly increased the content of AST IV in the affected and the healthy cerebral cortex. The bioactive components of PNS such as ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 was mainly distributed in the affected side of the cerebellum when PNS was used alone. Borneol combined with AST IV + PNS significantly increased the content of ginsenoside Rb1 in the cerebral cortex, especially in the affected cortex, increased the content of Rg1 in the healthy and the affected cortex, and increased the content of notoginsenoside R1 in the cerebral cortex, especially in the affected cortex, as well as in the cerebellum. Conclusion AST IV and the bioactive components of PNS such as ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 have a certain distribution in the cerebral cortex and the cerebellum after cerebral ischemia-reperfusion in rats. AST IV was mainly distributed in the cerebral cortex when it was used alone, ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 were mainly distributed in the cerebellum when PNS was used alone. The combination of borneol combined with AST IV and PNS can promote the gather of AST IV, ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 to the cerebral cortex, especially to the cortex of the ischemia-reperfusion side; Moreover, it can promote the absorption of AST IV, ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 in the cerebral cortex to varying degrees, especially in the affected cortex.
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Objective: To establish an HPLC method for the simultaneous content determination of seven constituents in Compound Yi’anning Pills (CYP). Methods: The determination was performed on Shim-pack GIST C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid (gradient elution: 0-5 min, 5% acetonitrile; 5-12 min, 5%-12% acetonitrile; 12-17 min, 12%-20% acetonitrile; 17-30 min, 20% acetonitrile; 30-36 min, 20%-50% acetonitrile; 36-45 min, 50%-80% acetonitrile; 45-70 min, 80% acetonitrile) at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1, 220 nm for schisantherrin A, 250 nm for schisandrin, 268 nm for salvianolic acid B, and 270 nm for tanshinone IIA. The column temperature was set at 35 ℃ and the injection volume was 10 μL. Results: The linear range of detection concentration was 0.015-0.150 mg/mL for notoginsenoside R1, 0.077-0.766 mg/mL for schisandrin, 0.006-0.062 mg/mL for salvianolic acid B, 0.009-0.092 mg/mL for buddleodide, 0.050-0.503 mg/mL for ginsenoside Rg1, 0.067-0.670 mg/mL for ginsenoside Rb1, and 0.011-0.108 mg/mL for tanshinone IIA. Average recoveries respectively were 99.5%, 99.8%, 99.2%, 98.9%, 96.9%, 98.8%, and 97.1%. Conclusion: The method is simple, effective, and accurate, which can be used for simultaneous determination of seven active constituents in CYP.
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Objective: To study and optimize the preparation method of test solution from Panax notoginseng. Methods: The effects of extraction solvent, liquid-material ratio and extraction temperature on the optimization of the preparation method of test solution of notoginsenoside and ginsenosides in P. notoginseng were investigated by Box-Behnken response surface methology. The content of notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1 was simultaneously determined by HPLC. Results: The optimum method was as follow: 75% methanol was used for once reflux extraction, the ratio of liquid-material ratio was 1:40, the extraction temperature was 100 ℃. Conclusion: The optimum preparation of test solution is simple and feasible, and the extraction rate of components is high, which provides a reference for the preparation of test solution from P. notoginseng.
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Objective: To establish the determination and fingerprint of Gesang Jiangtang Capsules (GJC) for its quality control. Methods: UV spectrophotometric method was used for determining polysaccharides, total saponins, and total flavonoids of GJC. Ten batches of GJC were detected and recorded by UPLC. Similarity evaluation was performed by using Similarity Evaluation System for Fingerprint Chromatogram of TCM (2012) to confirm the common peaks. Results: In three batches of GJC, average polysaccharides content, average total saponins content, and average total flavonoids content were 4.56%, 2.97%, and 2.61%, respectively. UPLC fingerprints of 10 batches of GJC were established and 15 common peaks were confirmed. Puerarin, rutin, notoginsenoside R1, ginsenoside Rg1, ombuoside, ginsenoside Rb1, and ombuin were identified by chemical identification of the reference substance, corresponding to peaks 1, 5, 8, 10, 12, 14, and 15. Conclusion: The methods can be used for the quality control of GJC with good precision, accuracy, and reproducibility.
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Objective: To clarify the effect of solution environment on ultrafiltration separation of Panax notoginseng total saponins (PNS) based on the molecular state. Methods: In the experiment, the transmittance and surface tension were selected as indexes for analyzing the effect of ethanol, inorganic salts, surfactants, and pH on the molecular state of PNS. And then, ethanol, NaCl, and pH were selected as influencing factors to analyze the separation rule of notoginsenoside R1 (R1) and ginsenoside Rb1 (Rb1). Results: The intermolecular interaction force of saponins was weakened by increasing the ethanol concentration; The pH value promoted saponin ionization, increased critical micelle concentration, and increased PNS ultrafiltration transmittance; The salting out effect of inorganic salt reduced the critical micelle concentration and PNS transmittance; The surfactant type was related to the ultrafiltration separation behavior of PNS. Rb1 was more sensitive to the factors than R1 by response surface methodology. Conclusion: The effect of solution environmental factors on the ultrafiltration separation of PNS was clarified by the combination of single factor analysis and response surface methodology. And the saponins can be separated purposefully by dynamically adjusting the molecular state.
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This study aimed to investigate the effect of notoginsenoside R₁ in delaying H₂O₂-induced vascular endothelial cell senescence through microRNA-34a/SIRT1/p53 signal pathway. In this study， human umbilical vein endothelial cells(HUVECs) were selected as the study object; the aging model induced by hydrogen peroxide(H₂O₂) was established， with resveratrol as the positive drug. HUVECs were randomly divided into four groups， youth group， senescence model group， notoginsenoside R₁ group and resveratrol group. Notoginsenoside R₁ group and resveratrol group were modeled with 100 μmoL·L⁻¹ H₂O₂ for 4 h after 24 h treatment with notoginsenoside R₁(30 μmoL·L⁻¹) and resveratrol(10 μmoL·L⁻¹) respectively. At the end， each group was cultured with complete medium for 24 h. The degree of cellular senescence was detected by senescence-associated β-galactosidase(SA-β-Gal) staining kit， the cell viability was detected by cell counting kit-8， the cell cycle distribution was analyzed by flow cytometry， and the cellular SOD activity was detected by WST-1 method in each group. The expressions of SIRT1， p53， p21 and p16 proteins in HUVECs were detected by Western blot. In addition， the mRNA expressions of miRNA-34a， SIRT1 and p53 in HUVECs were assayed by Real-time PCR. These results indicated that notoginsenoside R₁ significantly reduced the positive staining rate of senescent cells， enhanced the cell proliferation capacity and intracellular SOD activity， decreased the proportion of cells in G₀/G₁ phase， and increased the percentage of cells in S phase simultaneously compared with the senescence model group. Moreover， notoginsenoside R₁ decreased the mRNA expressions of miRNA-34a and p53 and the protein expression of p53， p21 and p16.At the same time， notoginsenoside R₁ increased the protein and mRNA expressions of SIRT1. The differences in these results between the senescence model group and the notoginsenoside R₁ group were statistically significant(<0.05). However， there was not statistically significant difference in these results between the notoginsenoside R₁ group and the resveratrol group. In conclusion， the senescence of endothelial cells induced by H₂O₂ can be used as a model for studying aging. Notoginsenoside R₁ has an obvious anti-aging effect on vascular endothelial cells in this study. The possible mechanism is that notoginsenoside R₁ can delay the senescence process of vascular endothelial cells induced by H₂O₂ by regulating microRNA-34a/SIRT1/p53 signal pathway.
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Humans , Cells, Cultured , Cellular Senescence , Ginsenosides , Pharmacology , Human Umbilical Vein Endothelial Cells , Hydrogen Peroxide , MicroRNAs , Genetics , Signal Transduction , Sirtuin 1 , Genetics , Tumor Suppressor Protein p53 , GeneticsABSTRACT
Objective To establish the quality standard for Yiwei Xiaoyu Granules.Methods TLC was used for the qualitative analyses of Atractylodis Macrocephalae Rhizoma, Angelicae Sinensis Radix and Dendrobii Caulis; HPLC was applied to determine the contents of notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1and ginsenoside Re. Results The TLC spots were clear and free from negative interference. Notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1and ginsenoside Re showed good linear relationships within the ranges of 0.067 2–0.672 μg (r=0.999 6), 0.501 2–5.012 μg (r=0.999 6), 0.326 5–3.265 μg (r=0.999 6), 0.098 3–0.983 μg (r=0.999 7), whose average recoveries were 96.32% (RSD=1.3%), 96.45% (RSD=1.5%), 101.23% (RSD=1.7%), and 97.89% (RSD=1.7%), respectively. Conclusion This method is accurate, reliable, and reproducible, which can be used for the quality control of Yiwei Xiaoyu Granules.
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Sanqi in Chinese herbal medicine is the root and rhi-zoma of Panax notoginseng (Burk.)F.H.Chen. As the effects of strengthening with tonics, promoting blood circulation to re-move blood stasis,relieving swelling and pain and hemostasia,it is widely used as a tonic medicine in the traditional Chinese medicine. The main active constituents of Sanqi are panax noto-ginseng saponins,including ginsenoside Rg1,Rb1 and notogin-senoside R1. Notoginsenoside R1 is one of the unique monomer compositions of panax notoginseng,which is often used as an in-gredient indicator in new drug research and development.In cur-rent years, the scientists have been conducted tremendous fun-damental studies to research the pharmacological activities of no-toginsenoside R1, to reveal its protective effects on the cardio-vascular system,central nervous system as well as to many other aspects. It is hoped that the relevant study about the pharmaco-logical action of notoginsenoside R1 would help its further clini-cal application. This paper mainly reviews the research on the pharmacological mechanism of notoginsenoside R1 in recent years.
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AIM To investigate the pharmacokinetic behaviors of four constituents in sugar-free Xinnaosu Granules in rat plasma.METHODS Rats intragastrically administered with the 0.5％ CMC-Na suspension of this drug (3 g/kg) had their blood collected for the determination of plasma concentration by UHPLC-MS/MS,after which pharmacokinetic parameters were calculated by DAS2.0 software.RESULTS The plasma concentrationtime curves for tanshinone Ⅱ A,salvianolic acid B,ginsenoside Rg1,notoginsenoside R1 accorded with two compartment model,with the t1/2values of (1.68 ±0.56),(4.13 ±0.87),(3.62 ±0.87),(9.77 ±3.12) h,Tmax values of (0.51 ±0.19),(1 ±0),(6 ±0),(4.00 ±1.09) h,Cmax values of (0.42 ±0.08),(0.17 ±0.02),(0.46±0.11),(0.41 ±0.12) mg/L,respectively.CONCLUSION All the four constituents in sugar-free Xinnaosu Granules demonstrate high bioavailabilities.
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AIM To establish an HPLC method for the content determination of six constituents in Shiyifang Vinum (Notoginseng Radix et Rhizoma,Dipsaci Radix,Carthami Flos,etc.).METHODS The content determination of notoginsenoside R1,ginsenoside Rg1,ginsenoside Rb1 and asperosaponin Ⅵ was performed on a 30℃ thermostatic Inertsil(R) ODS-3 C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 203 nm.The content determination of brucine and strychnine was conducted on a 30 ℃ thermostatic Geminni(R) C18 110(A) column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-mixed solution of 0.01 mol/L sodium heptanesulfonate and 0.02 mol/L potassium dihydrogen phosphate flowing at 1.0 mL/min in an isocratic elution manner,and the detection wavelength was set at 260 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 98.52％-99.96％ with the RSDs of 2.0％-2.3％.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Shiyifang Vinum.
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Notoginsenoside R1(NGR1),a critical compound in traditional herb Panax notoginseng, is a kind of estrogen receptor agonist.It is reported to exhibit anti-apoptotic,anti-oxidative and anti-inflammatory properties activity, so it is widely used for treatment of various diseases.In order to investigate the potential neuroprotective effect of NGR1 in hypoxic-ischemic brain damage(HIBD), primary cortical neurons were used in this study to establish oxygen-glucose deprivation/reoxygenation(OGD/R) injury models. They were treated with NGR1 and estrogen receptor inhibitor ICI-182780 respectively, then the neuronal survival, cell membrane integrity and apoptosis were assessed by MTT assay,lactate dehydrogenase test(LDH) and Hoechst 33342 stain respectively, while the protein expression levels of ATF6α,p-Akt,Akt,Bax and Cleaved Caspase-3 were measured by Western blotting. Results indicated that as compared with the blank control group,OGD/R could induce cell injury and apoptosis(P<0.05), reduce relative integrity of cell membrane(P<0.05), decrease protein expression of ATF6α,p-Akt(P<0.05), and increase protein expression of Bax and Cleaved Caspase-3(P<0.05) in the primary cortical cells. After NGR1 treatment, the expression levels of ATF6α,p-Akt were obviously increased, and the expression levels of Bax and Cleaved Caspase-3 and the apoptosis of neuron were decreased(P<0.05). However, these neuroprotective properties of NGR1 against ODG/R-induced cell damage could be blocked by ICI-182780. This finding indicated that NGR1 may protect the primary cortical neurons against OGD/R induced injury,and the mechanism may be associated with accelerating the activation of the ATF6/Akt signaling pathway via estrogen receptors.
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Objective: To develop an HPLC-ELSD method for the simultaneous determination of six saponins constituents including notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd in Xinling pills.Methods: HPLC-ELSD was used, and the chromatographic separation was performed on an Agilent Eclipse XBD-C18 column (150 mm×4.6 mm,5 μm) with acetonitrile-water as the mobile phase with gradient elution at the flow rate of 1.0 ml·min-1, the column temperature was maintained at 20℃, the drift tube temperature was 60℃, and the gas pressure was 4.00 bar.Results: Notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd was linear within the range of 0.30-6.00μg(r=0.999 5), 1.14-22.80μg (r=0.999 6), 0.17-3.40 μg (r=0.999 7), 0.81-16.20 μg (r=0.999 7), 0.08-1.60 μg (r=0.999 8) and 0.07-1.40 μg (r=0.999 8), respectively.The average recovery was 98.23%, 97.98%, 99.14%, 99.15%, 98.72% and 98.37%, and the RSDs were 1.56%, 1.31%, 1.16%, 1.07%, 0.73% and 0.92%(n=6), respectively.Conclusion: The method is convenient, accurate and reproducible in the quality control of saponins components in Xinling pills
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Objective To establish the quality standard of Fufang-Shiwu-Zhixue powder. Methods The microscopical identification was adopted to analyze charred typha pollen and cuttlebone. TLC was used to identity rhubarb and radix notoginseng. UV-HPLC was used to determine the contents of notoginsenosides R1, ginsenosides Rg 1, ginsenosides Rb1. Results Microscopic identifications shower the characteristics of harred typha pollen and cuttlebone. The identified characteristics of rhubarb and radix notoginseng by TLC were distinct and the spots were clear. Notoginsenosides R1, ginsenosides Rg 1, ginsenosides Rb1 showed good linearity in the range of 0.16-1.58 μg (r=0.9998), 0.58-5.81 μg (r=0.9999), 0.33-3.29 μg (r=1.0000), respervtively.The average recoveries were 98.51% (RSD=1.81%), 97.80% (RSD=2.04%), 98.22%(RSD=1.45%). Conclusions The method is accurate, simple and repeatable, which could be used for the quality control of Fufang-Shiwu-Zhixue powder.