Neuroprotective effect of ginsenoside Re on drosophila model of Parkinson's disease / 中国中药杂志

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.

Quality evaluation of ginsenoside reference substances based on qNMR spectroscopy / 中国中药杂志

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.

Research on Ginsenoside Re inhibition of CFs proliferation by regulating Wnt/β-catenin signaling pathway / 国际中医中药杂志

Objective:To observe the effect of Ginsenoside Re on the proliferation and protein secretion of primary cardiac fibroblasts (CFs) cultured in high glucose by vitro, and the regulation of Wnt/β-catenin signaling pathway.Methods:The myocardial fibroblast proliferation model induced by high glucose in vitro was used. Cell proliferation was detected by MTT method, cell cycle was measured by flow cytometry, concentration of type Ⅰ,Ⅲ collagens and TGF-β 1 protein were tested by ELISA assay. Protein expression of β-catenin, GSK-3β and p-GSK-3β were determined by Western blot. Results:Compared with the model group, the cell proliferation in Ginsenoside Re high, medium, low group were significantly decreased ( P<0.01), the percentage of cells in G 0 + G 1 phase was increased ( P<0.01), and the percentage of cells in S + G 2 + M phase was decreased ( P<0.01), the content of TGF-β 1 was significantly decreased( P<0.01). The content of type Ⅲ collagen [(6.566±1.620)ng/ml,(7.170±0.470)ng/ml vs. (11.241±2.234)ng/ml] in Ginsenoside Re high, medium group were significantly decreased ( P<0.01). The expression of β-catenin (0.281±0.016, 0.301±0.021 vs. 0.409±0.037) was significantly decreased and the expression of p-GSK-3β (0.369±0.049 vs. 0.268±0.048) in Ginsenoside Re high, medium group were significantly increased ( P<0.01). Conclusion:Ginsenoside Re plays an important role in inhibiting CFs proliferation and reducting the synthesis of collagen and TGF-β 1 by regulating abnormal expression of Wnt/β-catenin signaling pathway. It has the potential to delay the myocardial fibrosis of diabetes mellitus.

Effects of ginsenoside Re on intestinal mucosa of mice with severe acute pancreatitis and its regulation mechanism / 解剖学报

Objective To explore the effect and mechanism of ginsenoside Re on the intestinal mucosa of severe acute pancreatitis (SAP) mice by regulating the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/ STAT3) pathway. Methods Totally 48 mice were divided into : control group, SAP group, SAP + ginsenoside Re group and SAP + ginsenoside Re + LY2784544 group (n = 12). The mice were intraperitoneally injected with caerulein solution (after fasting for 12 hours, 100 μg/kg, 6 times, injection interval 60 minutes) to establish SAP models. Mice in the SAP+ ginsenoside Re group were intraperitoneally injected with ginsenoside Re (4 mg/kg, 1 time a day for 7 consecutive days). Intraperitoneal injection of 12.7 mg/kg LY2784544 was used to inhibit the JAK2/STAT3 pathway. Pancreatic and intestinal mucosal injury were detected in each group. The wet to dry weight ratio of pancreas, serum amylase and inflammatory factor levels were detected in each group. The intestinal mucosal barrier function was analyzed by detecting the levels of D-lactic acid and fluorescein isothiocyanate dextran (FITC-D). The damage of pancreatic tissue and intestinal mucosa tissue was observed by HE staining. Western blotting was used to detect the levels of apoptotic proteins Bax and Bcl-2 in the intestinal mucosa. The JAK2/STAT3 pathway expression levels in pancreatic tissue and intestinal mucosa tissue were detected by Realtime PCR and Western blotting. Results The pancreatic index, serum amylase, interieukin 6 (IL-6) and tumor necrosis factor a (TNF-a), FITC-D, D-lactic acid, Bax protein levels, JAK2 and STAT3 mRNA and protein levels in the SAP group were significantly higher than those in the control group, while Bcl-2 protein was significantly lower than that in the control group (P<0.05). The Bcl-2 protein of SAP+ginsenoside Re group was significantly higher than that of SAP group, and other indexes were significantly lower than those in SAP group (P<0.05). The Bcl-2 protein of SAP+ginsenoside Re+ LY2784544 group was significantly higher than that of SAP + ginsenoside Re group, and other indexes were significantly lower than those in the SAP+ginsenoside Re group (P<0.05). Conclusion Ginsenoside Re may reduce the pancreatic injury in SAP model mice by inhibiting the JAK/STAT pathway to alleviate the inflammatory response, and may protect the small intestinal mucosal barrier by alleviating pancreatitis and inhibiting the intestinal mucosal JAK/STAT pathway to inhibit cell apoptosis.

Quality evaluation of different origins and commercial grades of Panax notginseng by HPLC and grey correlation analysis / 中草药

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.

Network pharmacologic molecular mechanism of Shenmai Injection in treatment of COVID-19 combined with coronary heart disease / 中草药

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.

Effects of temperature regulation on ginsenoside accumulation and expression of key enzyme genes / 中草药

Objective: To explore the effect of temperature regulation on the accumulation of ginsenosides and the expression of key enzyme genes in the synthetic pathway. Methods: The ginseng callus cultured for 23 d was used as the test material and placed in six incubators at 5, 10, 15, 20, 25, and 30 ℃. The dry fresh weight and saponin content were measured to determine the response sensitive temperature. GAPDH was used as the internal reference gene. Real-time PCR was used to detect nine key enzymes in the saponin synthesis pathway, 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR2), Farnesyl pyrophosphate synthase (FPS), squalene synthetase (SS1), squalene epoxidase (SE1), dammarane diol synthase (DS-II), β-xanthin synthase (PNY1), β-xanthophyll C-28 hydroxylase (CYP716A52v2), protopanaxatriol synthase (CYP716A53v2), and protopanaxadiol synthase (CYP716A47). Results: 20 ℃ was the optimal temperature for the accumulation of dry and fresh weight of ginseng callus. The content of saponin in 5, 10, and 15 ℃ reached the maximum value for 2-3 d. Re, Rg1, and total saponins were 1.93, 11.93, and 1.54 times that of the control group, respectively. The expression levels of HMGR2, SS1, DS-II, SE1 and CYP716A52v2 reached their maximum values at 2-4 d of low temperature, which were 2.8, 1.6, 3.5, 3.7, and 3.8 times higher than those of the control group. Correlation analysis found that SE1 was significantly positively correlated with Re, Rg1 and total saponins. Conclusion: Moderate low temperature is conducive to the rapid accumulation of ginsenosides. SS1, DS-II, SE1, HMGR2, and CYP716A52v2 are key genes that respond to low temperatures, which play an important role in the process of ginsenoside biosynthesis against low temperatures.

Effect of particle design on pharmacokinetics of Shenling Baizhu Pulvis in rats / 中草药

Objective: Using LC-MS to explore the pharmacokinetic process in rats of Shenling Baizhu Pulvis (SBP), which was modified by particle design technology. Methods: Particle design powder of SBP was prepared by particle design technology. A scientific and feasible LC-MS analysis method was established to determine the blood concentration of index compounds such as ginsenoside Re (GI-Re), ginsenoside Rb1 (GI-Rb1), ginsenoside Rg1 (GI-Rg1), atractylenolide I (AT-I), atractylenolide II (AT-II) and pachymic acid (PA) in rats at different time points after administration. DAS 3.2.8 pharmacokinetic software was adopted to analyze the data, which related to blood concentration of index compounds, and the pharmacokinetics parameters were calculated by the non-compartmental model. Results: LC-MS analysis method was established, which has a good linear relationship and specificity for the index compounds in rats, and the RSD of precision, accuracy, extraction recovery and stability were all less than 5% or 10%. Compared with ordinary powder, the particle design powder displayed increased Cmax and AUC0-∞ after administration, and the AUC0-∞ of GI-Re, GI-Rb1, GI-Rg1, AT-I, AT-II and PA were increased to 1.52, 2.02, 1.22, 1.41, 1.13 and 1.43 times, respectively. Conclusion: The LC-MS analysis method meet the requirements of biological sample analysis in Pharmacopoeia of the People's Republic of China. After particle design and modification, the absorption speed of SBP in vivo become faster and the bioavailability is improved significantly.

Determination of ginsenoside Rg1 and Re in Tongru granules by HPLC / 国际中医中药杂志

Objective To establish a rapid and efficient method for the simultaneous determination of ginsenoside Rg1 and ginsenoside Re in Tongru granules by HPLC.Methods The HPLC analysis was carried out with Agilent ZORBAX SB-C18 and acetonitrile-0.1％ phosphoric acid(15:85,V/V) as mobile phase,the flow rate was 1.0 ml/min,the column temperature was 30 ℃,injection volume was 10 μl,and the detection wavelength was 203 nm.Results The Rg1 solution of ginsenoside showed a good linear relation between 0.136-2.040 μg (r2=0.999 9).The ginsenoside Re solution presented a good linear relationship (r2=0.999 8)between 0.066-0.990 μg,with the average addition recovery of 99.3％ and 99.1％,and the relative standard deviation of 0.84％ and 0.75％(n=6),respectively.Conclusions The method is highly specific,sensitive,reproducible,simple and efficient which can be used for quality control of Tongru Granules.

Optimization of extraction process of Shenqi Qiangxin Tablets based on pharmacodynamic test and analytic hierarchy process / 中草药

Objective: To optimize the extraction technology of Shenqi Qiangxin Tablets (SQT). Methods: With the improvement of heart lesion of pharmacological model of isoproterenol induced heart failure in rats as the index, pharmacological efficacy test was used to screen extracting conditions of the technology. The extraction technology was optimized by analytic hierarchy process combined with principal component analysis, single factor and orthogonal tests using each content of solid matter, ginsenosides Rg1, Re as indexes. And the verification test was carried out by using solid mass and icariin content as indexes. Results: Pharmacological efficacy test showed that technology 4 was superior. The optimal extraction condition of technology 4 was as follow: five medicinal materials including red ginseng and astragalus were reflux extracted three times with 50% ethanol, 11 fold for the first time, 10 fold for the second and three times, 2.5 h for each extraction; Epimedium and the other two medicinal materials were decocted three times with water, 19 fold for the first time, 16 fold for the second and third times, 1.5 h for each decction. The verification test showed that the average yield of ethanol extracted solids was 19.78%, and the average extraction rate of ginsenoside Rg1 and Re was 77.52%; The average value of water extracted solids was 16.58%, and the average extraction rate of epimedium was 90.98% (RSD < 2.0%, n = 3). Conclusion: The optimized extraction technology was stable and feasible.

Effect of different decocting methods on active ingredients in Banxia Xiexin Decoction / 中草药

Objective To investigate the effects of the three methods of decocting with deslag, decocting without deslag, and double decocting on the content of nine ingredients baicalin, baicalein, ginsenoside Re, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, liquiritin, 6-gingerol, berberine hydrochloride, palmatine hydrochloride, and total flavonoids in Banxia Xiexin Decoction (BXD). Methods Nine index components were determined by HPLC. The HPLC analysis was performed on Welch Ultimate XB-C18 column (250 mm × 4.6 mm, 5 μm) with mobile phase of acetonitrile-0.1% phosphate aqueous solution for gradient elution; And carried out at column temperature of 28 ℃, volume flow of 0.9 mL/min, and detection wavelength of 203, 252, 280, and 355 nm. The total flavonoids were determined by colorimetry. Results Nine kinds of ingredients and total flavonoids could be detected in three different decoctions. In the method of decocting with deslag, baicalin, baicalein, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, and liquiritin increased by 10.01%, 12.88%, 29.09%, 16.75%, and 15.02%, respectively, compared with decocting without deslag; It decreased by 5.54%, 4.15%, 14.49%, 7.85%, and 9.18%, respectively compared with double decocting; Ginsenoside Re, 6-gingerol, berberine hydrochloride, and palmatine hydrochloride increased by 37.90%, 3.78%, 5.33%, and 5.99% compared with decocting without deslag, respectively; compared to the double decocting methods, it increased by 1.07%, 11.57%, 3.41%, and 1.93%. The total flavonoids increased 22.61% higher than decocting without deslag and 6.54% higher than double decocting. Conclusion: The results can effectively reflect the quality difference of different decocting methods. Among the three methods of decoction, the method of decocting without deslag has significantly improved the dissolution of the active ingredients of each component in the decoction, and improve the clinical efficacy of BXD to a certain extent. It provides a good experimental basis for the decocting without deslag method used in Zhang Zhongjing’s Treatise on Febrile Diseases.

Determination of seven saponins in flower buds of Panax ginseng by quantitative analysis of multi-components by single marker / 中草药

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.

Comparative Analysis of Ginsenosides in Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra by UFLC-QTRAP-MS/MS / 中国药学杂志

OBJECTIVE To develop a comprehensive analytical method based on UFLC-QTRAP-MS/MS for simultaneous determination of protopanaxadiol [ginsenoside Rb1, Rc, Rb2, Rd, F2, 20(S)-Rg3, 20(R)-Rg3, CK], protopanaxatriol [ginsenoside Re, Rg1, Rf, 20(S)-Rg2, 20(S)-Rh1, 20(R)-Rg2, 20(R)-Rh1, F1] and oleanolic(ginsenoside Ro) in Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra. METHODS Under the optimized chromatographic conditions, good separation for seventeen target compounds was obtained on a SynergiTM Hydro-RP 100 column(2.1 mm×100 mm, 2.5 μm) at 40 ℃ with 0.1% aqueous formic acid (A)/acetonitrile (B) as the mobile phase by gradient elution at a flow rate of 0.4 mL·min-1. The target compounds were analyzed under multiple reaction monitoring (MRM) mode with an ESI source operated in negative ion mode, and principal component analysis (PCA) and hierarchical cluster analysis (HCA)were used for data processing. RESULTS The calibration curves of the 17 components had good linearity (r>0.999 0). The precision, repeatability and stability were all satisfying.The average recoveries of standard addition for the compounds were between 96.69% and 102.01%,and the relative standard deviations were less than 5%. The results of PCA and HCA showed that Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra were clearly distinguished.The main compositions with significant difference were ginsenoside 20(S)-Rg3, 20(R)-Rg3, 20(S)-Rh1, 20(R)-Rh1, and 20(R)-Rg2. CONCLUSION The established method could provide a new technique for the comprehensive evaluation and quality control of Ginseng Radix et Rhizoma and Ginseng Radix et Rhizoma Rubra, at the same time, it would pave the way for discovering the material basis contributing to the different properties and efficacies of the two medicinal materials.

Simultaneous determination of six constituents in Yitai Capsules by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of ginsenoside Re,ginsenoside Rg1,ginsenoide Rb1,specnuezhenide,calycosin-7-O-β-D-glucoside and oleanolic acid in Yitai Cap sules (Ginseng Radix et Rhizoma,Astragali Radix,Ligustri lucidi Fructus,etc.).METHODS The analysis of 70％ ethanol extract of this drug was performed on a 25 ℃ thermostatic Luna C18 column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-O.2％ phosphoric acid flowing at 1.0 mL/minin a gradient elution manner,and the detection wavelength was set at 203 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 95.58％-102.12％ with the RSDs of 0.82％-1.73％.CONCLUSION This simple and stable method can be used for the rapid quality control of Yitai Capsules.

Study on Quality Standard of Yiwei Xiaoyu Granules / 中国中医药信息杂志

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.

Simultaneous Determination of Seven Components in Shenrong Guben Tablets by HPLC Wavelength Switching Method / 中国药师

Objective:To develop an HPLC wavelength switching method for the determination of seven components in Shenrong Guben tablets simultaneously.Methods:A Waters Sunfire C18column (250 mm ×4.6 mm,5 μm) was adopted. The column temperature was set at 30 ℃. The mobile phase was acetonitrile-0.1% phosphate solution with gradient elution. The flow rate was 0.9 ml·min-1. The detection wavelength were set at 330 nm for verbascoside and martinoside,230 nm for albiflorin and paeoniflorin,and 203 nm for ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1. Results:The linear range of verbascoside, martinoside,albiflorin, paeoniflorin, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1was 6.38-159.50 μg·ml-1(r =0.999 3),3.19-79.75 μg·ml-1(r =0.999 9),4.37-109.25 μg·ml-1(r =0.999 5),14.26-356.50 μg·ml-1(r =0.999 4), 1.95-48.75 μg·ml-1(r = 0.999 8), 2.21- 55.25 μg·ml-1(r = 0.999 7) and 2.09- 52.25 μg·ml-1(r = 0.999 1), respectively. The average recovery and the corresponding RSD was 98.24% (1.11%),97.64% (1.43%),99.23% (0.80%), 100.13% (0.65%),96.99% (1.56%),98.10% (1.24%) and 97.75%(1.37%),respectively. Conclusion:The developed method can determine the contents of verbascoside, martinoside, albiflorin, paeoniflorin, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1in Shenrong Guben tablets simultaneously,which can be applied in the quality control of Shenrong Guben tablets.

A Comparative Pharmacokinetic Study of Ginsenoside Re after Oral Administration in Normal and Ultraviolet B Irradiation-induced Damage Rats / 分析化学

A methodology of quantitative analysis on ginsenoside Re (G-Re) in rat plasma by ultra performance liquid chromatography-triple quadrupole mass spectrometry was developed for comparing the pharmacokinetic profiles between normal rats and Ultraviolet B (UVB) irradiation-induced damage rats after oral administration. The sample separation was carried out on an Ascentis?Express C18column (5.0 mm× 3.0 mm,2.7 μm) with 0.1% formic acid in water and acetonitrile as the mobile phase under gradient elution. MS analysis was operated in multiple-reaction monitoring (MRM) mode using electrospray ionization (ESI) with negative ion mode,and the ions for quantification were m/z 991.54/945.53/475.60. The limit of detection (LOD,S/N=3), limit of quantification (LOQ, S/N=10) were 4.0 ng/mL and 13.5 ng/mL, respectively. G-Re was in good linearity between 15 ng/mL and 20000 ng/mL(r=0.999),the intra-day and inter-day precisions, recovery, matrix effect and stability could meet the pharmacokinetic analysis requirement. The results indicated that the metabolic process of G-Re conformed to a two-compartment pharmacokinetic model after single oral administration in the normal and model groups. The t1/2αwere(0.21± 0.04) h and (0. 69 ± 0. 07) h, respectively; t1/2βwere (17. 08 ± 0. 53) and (21. 40 ± 16. 77) h, respectively;AUC(0-t)were (321.91±2.27) μg/(L·h) and (474.99±194.96) μg/(L·h), respectively;AUC(0-∞)were (332. 44 ± 1. 66) μg/(L·h) and (518. 64 ± 231. 39) μg/(L·h), respectively; the pharmacokinetic parameters were significantly different between normal and UVB irradiated rats (p<0.05), except for t1/2α. This UHPLC-QQQ-MS method showed excellent separation, accuracy, high sensitivity, specificity and good repeatability,and it was suitable for the pharmacokinetic study of G-Re in vivo.

The effect of Re on immune response against subunit anti-caries vaccie rPAc / 安徽医科大学学报

Objective To study the immune efficacy of ginsenoside Re as the adjuvant of anti-caries subunit vac-cine rPAc. Methods 40 mice were randomly diveded into four groups:anti-caries vaccine group( rPAc group) ,gin- senoside Re groups( Re group) , Re+rPAc group, normal saline group( NS group) . Mice were intranasally immu-nized twice on weeks 0 and 2 with rPAc,Re,Re+rPAc or normal saline,respectively. Concentration of specific anti-bodies in serum and saliva were detected by enzyme-linked immunosorbent assay(ELISA). In addition, cytokine production of spleen lymphocyte responses were also evaluated. 24 Wistar rats,infected with S. mutans Ingbritt, were randomly diveded into four groups:rPAc group,Re group, Re+rPAc group, NS group. Rats were jmmunized intranasally with rPAc,Re, Re+rPAc or normal saline,respectively. The Keyes method was used to determine the caries activity. Results The level of serum specific anti-PAc IgG, IgG1,IgG2a and saliva anti-PAc IgA were sig-nificantly higher in Re+rPAc group (P＜0. 05). In addition, the expression of cytokines interluekin-4 and inter-feron-γ were significantly upregulated in Re+rPAc group than those in any other groups(P＜0. 01). Re+rPAc im-munized rats showed significantly fewer E,Ds and Dm lesions than rPAc-immunized rats,Re-immunized rats or NS-immunized rats(P＜0. 05). Conclusion Re as the adjuvant of anti-caries subunit vaccine rPAc triggers a stronger humoral and cellular response against dental caries,and Re is a promising adjuvant for anti-caries vaccine rPAc.

Role of JAK2/STAT3 signaling pathway in ginsenoside Re pretreatment in prevention of isoproterenol-induced acute myocardial ischemia / 中国药理学通报

Aim To observe the protective effect of ginsenoside Re pretreatment on rats with isoproterenol-induced acute myocardial ischemia via JAK 2/STAT3 signaling pathway .Methods SD rat model with acute myocardial ischemia was established using isoprotere-nol.Seventy-five rats were randomly divided into five groups: control group, model group , puerarin group (PUE), high dose group (Re-H, 20 mg· kg -1) and Re low-dose group ( Re-L, 10 mg kg -1 ) .The blood flow on the heart surface of rats in each group was ob-served by moor laser blood flow imaging system .The levels of CK , LDH, SOD, MDA and GSH in myocar-dium were measured by ELISA .The expressions of Bax and Bcl-2 proteins were detected by immunohistochem-istry.The expressions of JAK , p-JAK, STAT3 and p-STAT3 proteins were detected by Western blot .Re-sults Compared with the control group , the mean blood flow on the heart surface of rats in the model group significantly decreased , the levels of CK , LDH and MDA in the myocardium increased , the levels of GSH and SOD decreased , the ratio of Bcl-2/Bax de-creased ( P <0.05 ) , and the expression of JAK 2/STAT3 pathway related proteins was enhanced ( P <0.05 ) . The mean blood flow on the heart surface markedly increased , the levels of CK , LDH and MDA decreased , the level of GSH-Px increased , the ratio of Bcl-2/Bax increased, and the expression of JAK2/STAT3 pathway proteins evidently increased in the Re-H group compared with those of the model group ( P<0.05 ) .Conclusion Ginsenoside Re pretreatment has a good protective effect on the myocardium in rats with acute myocardial ischemia , which may be related to the activation of JAK2/STAT3 signaling pathway .

Pharmacokinetic study of ginsenoside Re after vaginal administration in rabbits by UPLC-MS/MS determination / 中草药·英文版

Objective: A selective and sensitive ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was employed to study the pharmacokinetics of ginsenoside Re (GRe) in rabbits after vaginal administration of Xiaomi suppository to evaluate the systemic exposure of the suppository for the local treatment. Methods: Chromatographic separation was on an ACQUITY UPLC® BEH C18 column, and acetonitrile-0.1% formic acid was used as mobile phase in gradient elution. The plasma samples were deproteinized by acetonitrile, and the pharmacokinetic parameters were calculated by Winnonlin 6.4. Results: Calibration curve of GRe showed a good linearity over the concentration range from 5 ng/mL to 500 ng/mL (r = 0.9999). The low limit of quantification of 5 ng/mL could satisfy the experimental requirement. The intra-day and inter-day precision of GRe at three concentrations were less than 1.96%, and the average recoveries of GRe were more than 64.0%. The pharmacokinetic parameters for vaginal administration were as follows: Tmax, 0.5 h; Cmax, 20.88 ng/mL; AUC0-t, 64.71 h · ng/mL and the residence time was 3.06 h. By using deconvolution calculation method, the cumulative absorption fraction of GRe was about 0.89%. Conclusion: The systemic exposure of GRe was minimal after vaginal administration of Xiaomi suppository.