Diversity of Endophytes in Panax quinquefolium Root / 中国实验方剂学杂志

ObjectiveTo analyze the community structure of endophytes in Panax quinquefolium root and explore the dominant endophytic bacteria and fungi， to provide scientific basis for the establishment of endophytic microbial bank in P. quinquefolium root. MethodInternal Transcribed Spacer （ITS） sequencing and 16S sequencing were performed on six P. quinquefolium root samples collected from Wendeng， Shandong province on PacBio Sequel Ⅱ. ResultA total of 8 phyla， 11 classes， 23 orders， 27 families and 53 genera of endophytic bacteria were identified in P. quinquefolium root， among which an unidentified Burkholderiaceae and an unidentified Rhizobiaceae were dominant. A total of 9 phyla， 23 classes， 35 orders， 43 families and 48 genera of endophytic fungi were identified in P. quinquefolium root， among which an unclassified Helotiales and Pseudogymnoascus were dominant. The community structure of endophytic bacteria revealed that the roots were selectively enriched with nitrogen-fixing bacteria such as unidentified Rhizobiaceae， Bradyrhizobium and Herbaspirillum， which suggested that nitrogen is important for the growth of P. quinquefolium root. The community structure of endophytic fungi indicated that P. quinquefolium in Shandong province might be infected by unclassified Helotiales. ConclusionThere is a rich diversity of endophytic bacteria and fungi in P. quinquefolium root， which provides scientific basis for studying the interaction of the plant with endophytic microorganisms and screening the endophytes to promote the growth of P. quinquefolium root.

Resource components and utilization values of different parts of Panax quinquefolium in Shandong province / 中国中药杂志

To explore the resource components and availability of different parts of Panax quinquefolium in Shandong province, the paper employed the non-targeted metabolomics technology based on ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) to analyze the metabolites and their metabolic pathways in the root, fibril, stem, and leaf of P. quinquefolium. The content of seven ginsenosides and polysaccharides in different parts was determined by high performance liquid chromatography(HPLC) and ultraviolet-visible spectrophotometry(UV-Vis). The results showed that the metabolites were mainly sugars, glycosides, organic acids, amino acids and their derivatives, terpenoids, etc. The total abundance of metabolites followed the trend of leaf > root > fibril > stem. Most of the differential metabolites were concentrated in phenylpropane biosynthesis, flavonoid biosynthesis, citric acid cycle, and amino acid biosynthesis. The leaf contained high levels of sugars, glycosides, amino acids and their derivatives, and flavonoids; the root was rich in terpenoids, volatile oils, vitamins, and lignin; the fibril contained rich organic acids; and the stem had high content of nucleotides and their derivatives. The content of ginsenosides Re and Rb_1 was significantly higher in the root; the content of ginsenosides Rg_1, Rg_2, Rd, F_(11), and polysaccharide was significantly higher in the leaf; and the content of ginsenoside Rb_2 was significantly higher in the stem. We analyzed the resource components and availability of different parts of P. quinquefolium, aiming to provide basic information for the comprehensive development and utilization of P. quinquefolium resources in Shandong province.

Study on the grade standard and quality evaluation of Panax quinquefolium / 中国药房

OBJECTIVE To establish the grade s tandard for Panax quinquefoli um and to evaluate the quality of different grades of medicinal materials. METHODS Totally 24 batches of P. quinquefolium were used as test samples. Pearson correlation analysis method was used to analyze the correlation between qualitative analysis indicators （taproot length ，taproot diameter and weight of single root ）and internal component indicators （ethanol-soluble extract ，and the contents of ginsenoside Rg 1，ginsenoside Re ， ginsenoside Rb 1，ginsenoside Rc ，ginsenoside Rb 2，ginsenoside Rd ，pseudo-ginsenoside F 11）. Combined with chemometrics methods，the reference indexes for the classification of P. quinquefolium were selected ，and the classification standards were formulated. HPLC-ELSD fingerprints of 24 batches of P. quinquefolium were established and their similarity evaluation was also performed. The chromatographic peaks were identified by comparison with the reference substance ，and then the quality of different grades of P. quinquefolium was evaluated by cluster analysis. RESULTS After screening ，taproot diameter ，the weight of single root and the content of ginsenoside Rd were taken as the reference indexes for the classification of P. quinquefolium . According to above 3 indexes，P. quinquefolium were divided into 3 grades：special grade ，first grade and second grade. According to the center value of K-means clustering ，the total score of special-grade medicinal materials was more than 135.40，that of first-grade medicinal materials was 61.82-135.40，and that of second-grade medicinal materials was less than 61.82. In the HPLC-ELSD fingerprints of 24 batches of P. quinquefolium ，25 common peaks were confirmed ，and 7 characteristic peaks were identified. The similarity of the chromatograms of P. quinquefolium of special grade ，first grade and second grade with fingerprints ranged 0.980-0.989，0.962-0.968，0.940-0.949，respectively. The results of cluster analysis showed that different grades of P. quinquefolium could be identified significantly. CONCLUSIONS The grade standard and HPLC-ELSD fingerprints of P. quinquefolium are established，which can be applied for exclusive identification of P. quinquefolium ，and provide reference for its quality control and grade classification.

Research progress on chemical constituents, biological activities, quality evaluation, and product development of Panax quinquefolium / 药学学报

italic>Panax quinquefolium L. is a valuable Chinese herbal medicine with a large market demand. It has a complex chemical composition and numerous biological activities. At present, research on P. quinquefolium is focused on its underground parts, with especial interest in its saponins. There are few studies on non-saponins and the aboveground parts of Panax quinquefolium. Current quality standards are based on the saponin contents, which does not address the other benefits of Panax quinquefolium. This paper summarizes progress on the chemical components, pharmacological effects, quality evaluation, and product development of P. quinquefolium in recent years, and provides references for its further R&D and comprehensive utilization.

Microbial inoculants and garbage fermentation liquid reduced root-knot nematode disease and As uptake in Panax quinquefolium cultivation by modulating rhizosphere microbiota community / 中草药·英文版

Objective: To find a suitable ecological cultivation measure to solve the problem of root-knot nematode disease of Panax quinquefolium (Panacis Quinquefolii Radix) and the heavy metals accumulating in its roots. Methods: Three-year-old P. quinquefolium was treated with four different combinations of microbial inoculant (MI) and garbage fermentation liquid (GFL) [the joint application of ‘TuXiu’ MI and Fifty potassium MI (TF), the combination use of ‘No. 1′ MI and Fifty potassium MI (NF), ‘Gulefeng’ poly-γ-glutamic acid MI (PGA), GFL], and the untreated control (CK). Here, high-throughput sequencing, ICP-MS and UPLC were employed to systematically characterize changes of microbial diversity and structure composition, heavy metals (As, Cd and Pb) content and ginsenoside content among different treatments. Results: The results revealed that different MIs and GFL could increase the root dry weight of P. quinquefolium, PGA enhanced it by 83.24%, followed by GFL (49.93%), meanwhile, PGA and GFL were able to lessen root-knot nematode disease incidence by 57.25% and 64.35%. The treatment of PGA and GFL can also effectively reduce heavy metals in roots. The As content in GFL and PGA was decreased by 52.17% and 43.48% respectively, while the Cd and Pb contents of GFL and PGA was decreased somewhat. Additionally, the content of total ginsenosides was increased by 42.14% and 42.07%, in response to TF and NF, respectively. Our metagenomic analysis showed that the relative abundance of particular soil microbial community members related to the biocontrol of root-knot nematode disease and plant pathogen (i.e., Chaetomium in NF, Xylari in GFL, and Microascus in PGA), heavy metal bioremediation (Hyphomacrobium in PGA and Xylaria in GFL), and nitrogen fixation (Nordella and Nitrospira in TF) was significantly increased; notably, potential harmful microflora, such as Plectosaphaerella and Rhizobacter, were more abundant in the control group. Conclusion: MI and GFL could improve the quality of P. quinquefolium by modifying its rhizosphere microbial community structure and composition, both of them are beneficial to the development of ecological cultivation of P. quinquefolium.

Research progress on saponins, biological activities of saponins, and quality control of Panax quinquefolium / 中国中药杂志

Panax quinquefolium, as a common precious medicinal plant, has complex chemical components and unique pharmacological activities, which can play a healthcare role in the human body. With the deepening of research, the application of P. quinquefolium has become increasingly extensive. This paper summarized the research progress of the saponins isolated and identified from diffe-rent parts of P. quinquefolium, the structural classification and pharmacological activities of the saponins, and the quality control of Panacis Quinquefolii Radix. Further, this paper put forward the urgent problems to be solved in the development of P. quinquefolium. It is hoped to lay a foundation for the further study and provide reference for the research direction of P. quinquefolium.

Effects of magnesium supply level on growth, nutrient element absorption and distribution, and quality of Panax quinquefolium / 中国中药杂志

This study aims to investigate the effects of different magnesium supply levels on the growth, nutrient absorption and distribution, and quality of Panax quinquefolium, and to determine the optimum content of exchangeable magnesium in soil. Three-year-old plants of P. quinquefolium were used in this study, and eight magnesium supply gradients(CK, Mg1-Mg7) were designed for indoor pot experiment(cultivation in soil). The plant growth indexes, nutrient element content in soil and plant, and root saponin content were determined at the end of the growth period. The correlation analysis of nutrient element content in aboveground and underground parts of P. quinquefolium showed significantly negative correlations of magnesium-calcium, magnesium-potassium, and magne-sium-manganese. With the increase in magnesium supply level, the biological absorption coefficient of magnesium increased, while that of total nitrogen, potassium, iron, and manganese decreased; the biological transfer coefficient of magnesium decreased, while that of nitrogen, phosphorus, calcium, iron, and manganese increased. The saponin content was analyzed by principal component analysis, which showed the comprehensive score in the order of Mg4(2.537), Mg2(1.001), Mg3(0.600), Mg1(0), Mg7(-0.765), CK(-0.825), Mg6(-0.922), and Mg5(-1.663). The partial least squares-path modeling(PLS-PM) showed that the correlation coefficients of exchangeable magnesium and pH with quality were-0.748 and-0.755, respectively, which were significant. Magnesium-calcium, magnesium-potassium, and magnesium-manganese showed antagonism in the nutritional physiology of P. quinquefolium. Excessive application of magnesium can lead to the imbalance of nutrient elements in P. quinquefolium. The content of exchangeable magnesium in soil suitable for the quality formation of P. quinquefolium was 193.34-293.34 mg·kg~(-1). In addition to exchangeable magnesium, pH was also important to the quality formation of P. quinquefolium. Therefore, exchangeable magnesium and pH could be regarded as monitoring factors for the quality formation of P. quinquefolium.

Effects of Dimethyl Sulfoxide and Ethanol on Adventitious Root Growth of Panax quinquefolium and Ginsenosides / 中国实验方剂学杂志

Objective:To screen out the suitable nonpolar molecular cosolvent and concentration with adventitious root phenotype and ginsenoside content in the controlled experiment as the evaluation indexes， so as to lay a solid foundation for exploring the causes for good shape and high quality of <italic>Panax quinquefolium</italic>. Method:After being treated with different concentrations of dimethyl sulfoxide （DMSO） and ethanol， the adventitious roots were scanned using a panoramic scanner， and the resulting images were used for measuring the branch number and average diameter by WinRHIZO Pro 2016， Synbiosis ProtoCol 3 colony counter， Image J， and SmartRoot. The contents of ginsenosides Rg₁， Rb₁， and Re were determined by ultra-performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS）. Result:Compared with the blank control， the 0.1% DMSO and 75% ethanol made the adventitious root phenotype and ginsenoside contents significantly changed. Specifically， the branch number and average diameter were significantly reduced. The ginsenoside Rg₁ in the adventitious roots decreased after 0.1% DMSO treatment， whereas the ginsenosides Rg₁ and Re increased after 75% ethanol treatment. The adventitious root phenotype and ginsenoside contents in the 0.1% DMSO treatment group were not significantly different from those in the control group. Conclusion:The 0.01% DMSO does not affect the adventitious root growth of <italic>P. quinquefolium </italic>and is insoluble in water， enabling it to be considered as a suitable nonpolar molecular cosolvent for future research on the genetic causes for the good shape and high quality of <italic>P. quinquefolium</italic>.

Panax quinquefolium saponin Optimizes Energy Homeostasis by Modulating AMPK-Activated Metabolic Pathways in Hypoxia-Reperfusion Induced Cardiomyocytes / 中国结合医学杂志

OBJECTIVE@#To investigate the effects and underlying mechanisms of Panax quinquefolium saponin (PQS) on energy deficiency in hypoxia-reperfusion (H/R) induced cardiomyocytes.@*METHODS@#The H/R injury involved hypoxia for 3 h and then reperfusion for 2 h. Cardiomyocytes recruited from neonatal rat ventricular myocytes (NRVMs) were randomly divided into control, H/R, H/R+compound C (C.C), H/R+PQS, and H/R+C. C+PQS groups. BrdU assay, lactase dehydrogenase (LDH) leakage and early apoptosis rate were evaluated to assess cell damages. Contents of high energy phosphate compounds were conducted to detect the energy production. Protein expression levels of adenosine monophosphate-activated protein kinase a (AMPKα), glucose transporter 4 (GLUT4), phosphate fructose kinase 2 (PFK2), fatty acid translocase/cluster of differentiation 36 (FAT/CD36), and acetyl CoA carboxylase 2 (ACC2) in the regulatory pathways were measured by Western blotting. Immunofluorescence staining of GLUT4 and FAT/CD36 was used to observe the mobilization of metabolic transporters.@*RESULTS@#PQS (50 mg/L) pretreatment significantly alleviated H/R-induced inhibition of NRVMs viability, up-regulation of LDH leakage, acceleration of early apoptosis, and reduction of energy production (P<0.05). Compared with the H/R group, up-regulated expression of AMPKα, GLUT4, PFK2, FAT/CD36 and ACC2 were observed, and more GLUT4 and FAT/CD36 expressions were detected on the membrane in the H/R+PQS group (P<0.05). These effects of PQS on H/R-induced NRVMs were eliminated in the H/R+C.C+PQS group (P<0.05).@*CONCLUSION@#PQS has prominent advantages in protecting NRVMs from H/R-induced cell damages and energy metabolic disorders, by activation of AMPKα-mediated GLUT4-PFK2 and FAT/CD36-ACC2 pathways.

Establishment of Two-reference Substances for Determination of Multiple Components for 8 Kinds of Ginsenosides in Panax quinquefolium Broken Pieces / 中国药房

OBJECTIVE：To establish the m ethod for simultaneous determination of 8 kinds of ginsenosides in Panax quinquefolium broken pieces. METHODS ：HPLC-DAD method was used to determine the contents of ginsenoside Rg 1，Re，Rb1， Rc，Ro，Rb2，Rb3，Rd in P. quinquefolium broken pieces. The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid water solution （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelength was set at 203 nm，and sample size was 10 μL. Ginsenoside Re and ginsenoside Rb 2 were used as control ，liner calibration with two-reference substances correction was used to predict the retention time of other 6 components，and was compared with the relative retention time method. Using ginsenoside Re as control ， above components were quantified by the relative correction factor method ，and the results were compared with the external standard method. RESULTS ：The contents of ginsenoside Rg 1，Re，Rb1，Rc，Ro，Rb2，Rb3，Rd were 10.59-12.78，2.160-2.768， 27.492-38.880，3.154-4.018，3.368-4.080，0.343-0.755，0.961-1.415，5.857-6.923 mg/g. The accuracy of two-reference substances linear correction method to predict the retention time of components was higher ，and the absolute deviation of the predicted retention time was lower than that of the relative retention time method. There was no significant difference between the relative correction factor method and the external standard method ，and relative error was ＜3％ . CONCLUSIONS ：Established two-reference substances for determination of multiple components can be used for qualitative and quantitative analysis of 8 kinds of ginsensides in P. quinquefolium broken pieces simultaneously and accurately.

Molecular identification of Sanqi Tablets using DNA barcoding and its methodological evaluation / 中草药

Objective: In order to study the application of DNA barcoding in the authentication of Chinese patent medicines, Sanqi Tablets were used as the object to investigate the applicability, specificity and precision of this method. Methods: Fifteen batches of commercially available Sanqi Tablet samples were collected. The conditions of DNA extraction for Sanqi Tablet had been first investigated, and the DNA was used for testing the applicability of the methods such as PCR amplification, sequence acquisition, and species authentication in the principles for molecular identification of traditional Chinese materia medica using DNA barcoding. The specificity and reproducibility of DNA barcoding in identification of Sanqi Tablets and its adulterations from the roots of Panax notoginseng, P. ginseng and P. quinquefolius were also studied. Results: The Sanqi Tablet sample with an amount of sampling to be 100 mg and a water bath at 56 ℃ for 8 h gave an average concentration of 60.7 ng/μL and then the PCR amplification, sequence acquisition and species assignment were all successful. The ITS2 sequences of P. notoginseng, P. ginseng and P. quinquefolius were all 230 bp in length, and there were seven stable SNP loci between P. notoginseng and P. ginseng, P. notoginseng and P. quinquefolius. ITS2 sequences could be successfully obtained from lab-made and the adulterated Sanqi Tablets, and the Sanger sequencing chromatograms of different ratios of P. notoginseng and P. ginseng mixtures, P. notoginseng and P. quinquefolius mixtures had heterozygous peaks with corresponding peak height ratio at SNP positions. The repeatability, intermediate precision and reproducibility were all in line with the requirements of “General Regulation 9101” in the Chinese Pharmacopoeia. Conclusion: The ITS2 sequence can stably and accurately authenticate the raw materials of Sanqi Tablets with substantial specificity and precision. The DNA barcoding identification method of Sanqi Tablets will provide a new technical tool for ensuring the safety of Sanqi Tablets in clinical medications, and provide reference for the identification of other single-herb products documented in the Chinese Pharmacopoeia.

Study on the Heart Protective Effect of Panax quinquefolium Extract Based on Zebra Fish Model / 中国药房

OBJECTIVE：To study the heart protective effect of Panax quinquefolium extract of Zebra fish. METHODS ：The drug dosage of P. quinquefolium extract and the optimal modeling dose of terfenadine were determined by tolerance test ；24 hours post fertilization （24 hpf），Zebra fish were divided into control group （0.5％ dimethyl sulfoxide ），model group （2.5 μg/mL terfenadine），positive control group （2.5 μ g/mL terfenadine + 30 μ g/mL resveratrol） and different mass concentration P. quinquefolium extract groups （2.5 μg/mL terfenadine+5，10，25 μg/mL P. quinquefolium extract，by raw material ）. 48 h after incubation，the heart rate and sinus venous-bulbus arteriosus （SV-BA）interval of Zebra fish were used as indexes ，and the heart injury repair rate was calculated to investigate the heart protection of P. quinquefolium extract. Another 24 hpf Zebra fish were divided into blank control group ，model group ，positive control group ，20 batches of P. quinquefolium extract from different producing area groups （5 μg/mL，by raw material ），with same administration route and 48 h of incubation. The heart rate ，SV-BA interval of heart was determined ，heart injury repair rate and heart injury relative repair rate were calculated ；the heart protective effect of P. quinquefolium extract was further validated. RESULTS ：Compared with blank control group ，heart rate was decreased significantly（P＜0.01），while SV-BA interval was increased significantly in model group （P＜0.01）. Compared with model group，heart rate of Zebra fish was increased significantly in positive control group and 5，10，25 μg/mL P. quinquefolium extract groups（P＜0.01），while SV-BA interval of Zebra fish was shortened significantly in 5，10 μg/mL P. quinquefolium extract groups （P＜0.05 or P＜0.01）；heart injury repair rate of Zebra fish in 5 μg/mL P. quinquefolium extract group reached 73.77％. In validation test ，compared with model group ，heart rate was increased significantly （P＜0.01），while SV-BA interval of Zebra fish was decreased significantly in 20 batches of P. quinquefolium extract groups （P＜0.01）；heart injury repair rate ranged 70.45％-85.78％，and heart injury relative repair rate ranged 75.48％-98.12％. CONCLUSIONS ：P. quinquefolium extract has good heart protective effects.

Preliminary investigation on arbuscular mycorrhiza fungi of cultivated Panax quinquefolium roots / 中国中药杂志

In this study, the infection of root arbuscular mycorrhizal fungi(arbuscular mycorrhizal fungi, AMF of Panax quinquefolium in Shandong province was investigated, and the distribution characteristics and infection regularity of AMF were found out. The AMF of P. quinquefolium roots in different habitats was examined by alkali dissociation-trypickin blue staining method to study the infection rate and infection intensity. The contents of ginsenoside(Rb_1, Re, Rg_1, Rb_2, Rd and Rh_1) in the roots of P. quinquefolium was determined by HPLC. The experimental data were SPSS 17.0 statistical software for One-way analysis of variance, cluster analysis and correlation analysis. The results showed that the AMF infection in roots of P. quinquefolium, and there were obvious structures such as hyphae, arbuscular branches and vesicles, and the AMF infection rate and infection intensity showed obvious spatial and temporal heterogeneity with the growth age and origin of P. quinquefolium. The infection rate of AMF in roots of P. quinquefolium from 1 to 3 years increased significantly with the increase of growth years(P<0.05). The infection intensity and infection rate of P. quinquefolium showed a similar change trend, the AMF infection rate and infection intensity reached the highest level in the third year. Cluster analysis showed that the infection rates of roots of P. quinquefolium in similar geographical locations could be clustered together. Correlation analysis showed that the AMF infection rate of P. quinquefolium root was significantly positively correlated with the infection intensity, and the AMF infection rate and infection intensity were significantly positively correlated with the contents of ginsenoside Rg_1, Re and Rb_1. This study explored the distribution characteristics and regularity of AMF in roots of P. quinquefolium under the protected cultivation conditions, and provided basic data for ecological cultivation of P. quinquefolium and research and development of biological bacterial fertilizer.

Seeds identification of Panax ginseng and Panax quinquefolium using ITS2 DNA barcordes / 中草药

Objective Traditional identification methods of pharmacognosy is difficult to distinguish the seeds of Panax ginseng and Panax quinquefolius. In order to improve the efficacy and accuracy of identification and provide the scientific foundation for the establishment of Chinese herbal medicine seed quality standards, molecular identification methods of the seeds were established by DNA barcoding technology. Methods The pharmacognostical identification method was used to study the morphological identification and microscopic characters of different seeds. DNA barcodes and Chinese Pharmacopoeia species standard barcode database were employed to identify the seeds by ITS2 sequence comparison, genetic distance comparison and systematic NJ tree construction. Results Intraspecific genetic distances of individuals participating were smaller than interspecific genetic distances. Phylogenetic tree map showed that two species were repectively clustered into one. A total of 42 samples of seeds from Panax ginseng and Panax quinquefolius produced by nine areas were all top-quality and easy to distinguish. Conclusion ITS2 DNA barcodes can identify and differentiate the seeds of P. ginseng and P. quinquefolius germplasm resources quickly, accurately and efficiently.

Quality ecotype of Panax quinquefolium L. based on heredity-chemistry-ecology characteristics / 药学学报

Medicinal materials in China differ in quality by different ecological types. Our research group found that there were two ecotypes of domestic Panax quinquefolium L. according to the characteristics of ginsenosides, inside versus outside Shanhaiguan. The genetic and ecological mechanisms of quality variation of Panax quinquefolium L. is unknown. Based on the genetic-chemical-ecological strategy, transcriptome and HPLC technology were used for comprehensive correlation analyses of transcriptomic data, ginsenoside content and environmental climate ecological factors. The transcriptomic results showed that key genes of ginsenoside biosynthesis, such as HMGR, AS and FPS, were significantly down-regulated in the inside Shanhaiguan ecotype. HPLC results showed that the quality of outside Shanhaiguan ecotype Panax quinquefolium L. was higher than that of the inside ecotype, with the content of ginsenosides in outside Panax quinquefolium L. was higher than that of inside ecotype except Rb2. Correlation analyses revealed that content of Panax quinquefolium L. ginsenoside is positively related to the expression levels of ginsenoside biosynthesis key genes (MK, HMGS, HMGR, and AS), and negatively related to the expression of glycosyl transferase (GT). The content of ginsenosides is negative related with climate factors, such as temperature, sunshine, and is positively related with moisture in both ecological environments. This study has provided a new mechanistic insight into the quality variations of two ecotypes for Panax quinquefolium L. and established a scientific basis for studying the ecological factors for the quality of traditional Chinese medicine.

Determination of ginsenoside Re, pseudo-ginsenoside F11, ginsenoside Rb3, and ginsenoside Rd from flower buds of Panax quinquefolium / 中草药

Objective To establish the determination method of four ginsenosides from flower buds of Panax quinquefolium by Waters Acquity UPLC H-Class with Xevo TQD. Methods The chromatographic separation was performed on a Waters Acquity UPLC BEH C18 column (50 mm × 2.1mm, 1.7 μm). The mobile phase was a mixture of acetonitrile and water containing 0.05% ammonium hydroxide with gradient elution; The flow rate was 0.45 mL/min, and the column temperature was 35 ℃; Multiple reaction monitoring (MRM) acquisition under negative ion scan mode by ESI was also performed. Results The method was established with good precision, stability, repeatability, and accuracy. Ginsenoside Re, pseudo-ginsenoside F11, ginsenoside Rb3, and ginsenoside Rd showed a good linearity in the range of corresponding concentrations. Conclusion The UPLC-MS/MS method is rapid, simple, accurate, reliable, which can be used for the analysis of ginsenosides from flower buds of P. quinquefolium.

Effect of Panax Quinquefolium 20s-Protopanaxtriolsaponins on the Inflammatory Reaction after Cerebral Is-chemia Reperfusion Injury in Rats / 中国药师

Objective:To evaluate the effect of panax quinquefolium 20s-protopanaxtriolsaponins ( PQTS) on the inflammatory reac-tion after cerebral ischemia/reperfusion injury in rats.Methods: Wistar rats were randomly divided into the sham , the model, PQTS treatment groups respectively at dose of 12.5, 25.0 and 50.0 mg· kg-1 and the positive control group (flunarizine hydrochloride injec-tions, FHI, 2.0 mg· kg-1).All the rats were with intraperitoneal injection once a day for 3 consecutive days.The rats were subjected to 2-hour transient middle cerebral artery occlusion (MCAO) followed by 24-hour reperfusion.The neurological function was scored and the infarct volume was measured after the 24-hour reperfusion.Brain edema was measured by the dry-wet weight.Myeloperox-idase ( MPO) activity was determined by a spectrophotometer .The permeability of blood brain barrier was evaluated by the Evans blue content in brain determined by a spectrophotometer .The mRNA levels of interleukin-1β(IL-1β), interleukin-6 (IL-6) and tumor necrosis fac-tor-α( TNF-α) were evaluated by QPCR test .Moreover , the expression of NF-κBp65 was analyzed using Western blotting .Results:Compared with the model group , PQTS and FHI treatment groups (12.5, 25.0 and 50.0 mg· kg-1 ) and FHI group could improve the neurological function , decrease the infarct size , reduce the brain water content , inhibit the MPO activity and reduce the permeability of blood brain barrier.In addition, PQTS and FHI treatment groups (12.5, 25.0 and 50.0 mg· kg-1 ) also effectively inhibited the in-crease in the mRNA levels of IL-1β, IL-6 and TNF-α, and reduced the protein expression of NF-κBp65 when compared with the model group (P<0.05 or P<0.01).Conclusion:The results indicated that PQTS and FHI can significantly protect brain against cerebral is-chemia/reperfusion injury in rats by anti-inflammatory actions.

Extraction of Eight Ginsenosides from Leaves of Panax quinquefolium L. By Ultrasoinc Fountain-based Ultrasonic-assisted Nebulization Extraction Coupled with Solid Phase Extraction / 分析化学

A new ultrasonic-assisted extraction (UANE) method coupled with solid phase extraction (SPE) using ultrasonic fountain was established for the extraction of eight common ginsenosides from leaves of Panax quinquefolium L. The extraction system has been designed and several experimental parameters,including the type and volume of extraction solvent,pH value and salt concentration of extraction solvent,type and volume of elution solvent,and amount of C18, extraction time were examined and optimized. Under the optimal conditions,the recoveries of ginsenosides were in the range of 96. 3% -110. 6%, the relative standard deviations (RSDs) were in the range of 2.8%-4.3%,indicating that the method has a good performance for the extraction of these ginsenosides. Compared with traditional UANE-SPE method, the modified method simplified the extraction device,shortened the extraction time and improved the extraction efficiency.

Influence of Panax quinquefolium saponins on oxidative damage and function of vascular endothelium in diabetic rats / 中国药理学通报

Aim To investigate the protective effect of Panax quinquefolium saponins (PQS ) on oxidative damage and function of vascular endothelium. Meth-ods After 3 days of adaptive feeding of Wistar male rats,56 rats were randomly selected as the control group,while others were induced by streptozotocin (STZ,30 mg·kg - 1 ). The diabetic rats were random-ly divided into three groups:the model group,PQS group(100 mg · kg - 1 )and Vitamin E group (100μmol·kg - 1 ). The normal group was injected with e-qual amount of citrate buffer. PQS and Vitamin E groups of diabetic rats were administered orally once a day. After 4,8,16 weeks of administration,the con-centration of blood glucose and LPO,SOD of serum, heart and kidney were measured,and the tension of the aortic vascular ring was determined. Results Compared with model group,with prolongation of med-ication,the concentration of glucose and the LPO of serum,heart and kidney in the PQS group significantly decreased(P < 0. 05,P < 0. 01),then the results of the aortic vascular ring tension showed that acetylcho-line induced vasodilatation and maximum diastolic per-cent were obviously elevated in PQS group(P < 0. 05, P < 0. 01). Conclusion PQS could elevate the an-tioxidant function in diabetic rats,thus improving the endothelium-dependent vasodilation function and inter-fering with the occurrence and development of diabetic vascular complications

Identification of different ginsengs and producing areas of Panax quinquefolium using non-linear chemical fingerprint / 药学学报

Panax quinquefolium and Panax ginseng were investigated using non-linear chemical fingerprint technology, and a novel method to identify two ginsengs and different producing areas of P. quinquefolium was put forward. The non-linear chemical fingerprints of P. quinquefolium(collected from Canada, Jilin and Shaanxi)and P. ginseng (collected from Jilin)were obtained by reactions took place in the system of "H2SO4-MnSO4-CH3COCH3-NaBrO3" and their system similarities were evaluated. In the meantime, the quality difference in P. quinquefolium from different producing areas was evaluated using HPLC to determine the contents of main 7 ginsenosides. As a result, the non-linear chemical fingerprints exhibited a good reproducibility and characteristic when dosage of detection was 0.4 g and the reaction temperature was 37 ℃. The fingerprint characteristics of P. quinquefolium were obviously different from P. ginseng. The two species of ginsengs could be distinguished by the visual fingerprint characteristics, and P. quinquefolium from different producing areas were identified by the system similarities. Furthermore, HPLC analysis showed that the quality P. quinquefolium from different producing areas was varied, which indicated that rapid identification and quality evaluation of P. quinquefolium become very important and necessary. Compared with HPLC technology, non-linear chemical fingerprint is a more convenient, rapid and economic technology. This study provides a novel strategy to distinguish and evaluate P. quinquefolium and P. ginseng, which will provide a reference for the quality evaluation and control of Chinese medicine.