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The dry root and rhizome of Panax ginseng C. A. Mey has garnered much interest owing to its medicinal properties against diabetes and cardiovascular diseases. In this study, an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS)-based metabolomics approach was used to illustrate the therapeutic mechanisms of ginseng extract on the serum and urinary metabolic profiles in streptozotocin-induced type 1 diabetes mellitus (T1DM) rats. Pharmacological and renal parameters in response to the administration of ginseng were also evaluated. In total, 16 serum endogenous metabolites and 14 urine endogenous metabolites, including pyruvic acid, indoleacetic acid, and phenylacetylglycine, were identified as potential biomarkers for diabetes. Pathway enrichment and network analysis revealed that the biomarkers modulated by ginseng were primarily involved in phenylalanine and pyruvate metabolism, as well as in arginine biosynthesis. Moreover, the levels of several renal injury-related biomarkers in T1DM rats were significantly restored following treatment with ginseng. The administration of the extract helped maintain tissue structure integrity and ameliorated renal injury. The findings suggest that the regulatory effect of ginseng extract on T1DM involves metabolic management of diabetic rats, which subsequently attenuates T1DM-induced early renal dysfunction.
Subject(s)
Animals , Rats , Biomarkers , Chromatography, High Pressure Liquid/methods , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 1/drug therapy , Kidney , Metabolomics/methods , Panax/chemistry , Plant Extracts/pharmacologyABSTRACT
Panax japonicus, which in the Tujia dialect is known as “Baisan Qi” and “Zhujieshen”, is a classic “qi” drug of Tujia ethnomedicine and it has unique effects on disease caused by “qi” stagnation and blood stasis. This paper serves as the basis of further scientific research and development of Panax japonicus. The pharmacology effects of molecular pharmacology were discussed and summarized. P. japonicus plays an important role on several diseases, such as rheumatic arthritis, cancer, cardiovascular agents, and this review provides new insights into P. japonicus as promising agents to substitute ginseng and notoginseng.
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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.
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Objective:To analyze and identify ginseng miRNA. Methods: The miRNA and target genes of Shizhu ginseng and Yuan ginseng were detected by the degradation sequencing technology; Functional annotation of Degradome genes was carried out using public databases of KEGG/NR/GO database; The expression of miRNA and target genes of Shizhu ginseng and Yuan ginseng was determinated by real time fluorescence quantitative PCR technique. Results: A total of 13 target genes of eight miRNA families were obtained; The target gene type of miRNA was mainly transcriptional factor, response factor, and signal transduction pathway by means of KEGG/NR/GO database analysis. The results of real time fluorescence quantitative PCR verification of aqc-miR-159, bdi-miR162, cpa-miR319, pgi-miR4376, smo-miR396 and its target genes were basically consistent with the expression of miRNA and target genes from the degradation group. Conclusion: The target genes of partial Panax ginseng miRNA is clarified, which lays the foundation for further study of the possible function of ginseng miRNA.
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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.
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Objective: To study the chemical constituents of ginsenosides from the flower buds of Panax ginseng. Methods: The compounds were isolated and purified by Diaion HP-20, MCI gel, silica gel, and semi-preparative HPLC. The structures were elucidated based on NMR and MS data. Results: Four compounds were isolated from the extract of P. ginseng flower buds, and identified as 6’-acetyl-ginsenoside F1 (1), 12α-hydroxyl-ginsneoside Rd (2), 20(S)-ginsenoside Rg3 (3), and 5,6-didehydro-20(S)- ginsenoside Rg3 (4). Conclusion: Compound 4 is a novel ginsenoside, compounds 1 and 2 are new natural products.
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Objective: Human hepatoma SMMC-7721 cells were transplanted into nude mice to study the tissue distribution of nanostructured lipid carrier modified by hyaluronic acid (HA-OUR-NLC) loaded with three components in Panax ginseng (oleanolic acid, ursolic acid, and ginsenosider Rg3, OUR). Methods: FITC and DiR were used as fluorescent probes to dynamically monitor the HA-OUR-NLC targeted behavior of various tissues and organs through fluorescence endoscopic confocal imaging and in vivo imaging studies. Results: RUE values of oleanolic acid, ursolic acid, and ginsenosider Rg3 in tumors were significantly increased in HA-OUR-NLC group, reaching 2.51 ± 1.23, 2.27 ± 1.43, and 2.77 ± 0.25, respectively, which indicated that nanoparticles modified by hyaluronic acid could enhance drug uptake in tumors. The DiR accumulation in tumors of DiR-HA-OUR-NLC was higher than that of DiR-OUR-NLC by the visualized fluorescence of in vivo imaging. Conclusion: It indicated that nanoparticles modified by hyaluronic acid loaded with three components in P. ginseng can be enriched in the tumor site of liver cancer, which is in line with the expectation and can significantly improve the tumor targeting of the drug delivery system.
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Objective: To explain the phenomenon that Panax ginseng is not compatible with Raphani Semen based on pharmacodynamics and pharmacokinetics. Methods: The forced swimming time and biochemical parameters such as blood lactate (BLA), serum urea nitrogen (SUN), and hepatic glycogen (GLU) were determined for anti-fatigue experiment. The UPLC-MS/MS was used to analyze the pharmacokinetics of Rg1, Re, Rb1, and Rd after orally administration of P. ginseng and P. ginseng combined with Raphani Semen to rats. Pharmacokinetic differences of four ginsenosides between single uses of P. ginseng and combined with Raphani Semen were investigated. Results: The results showed that Raphani Semen tended to significantly reduce the anti-fatigue activity of P. ginseng. Co-administration of P. ginseng and Raphani Semen had significant effects on the pharmacokinetics of the four ginsenosides in rats compared to that observed with P. ginseng extract alone. The AUC0–12 h values of the four ginsenosides in PG group were higher than the corresponding values in the PR group. It can be inferred that Raphani Semen decreased the blood exposure of the four ginsenosides in rats when it combined with P. ginseng. Conclusion: The anti-fatigue activity and pharmacokinetic results showed that Raphani Semen may reduce the pharmacological actions of P. ginseng.
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OBJECTIVE： To establish a method for the determination of 33 kinds of pesticide residues in Panax ginseng C.A.Mey by GC-MS/MS and LC-MS/MS. METHODS： The 53 chemical monomers of 33 pesticide residues clearly prohibited by the Chinese ministry of agriculture were selected as the detection indicators. The samples were extracted with acetonitrile by high speed homogenizer. An LC-MS/MS analysis was performed on a CORTECSTM UPLC C18(2.1 mm×150 mm, 1.6 μm) column with isocratic elution of 0.1% formic acid (containing 5 mmol•L-1 ammonium formate) is mobile phase A, 95% acetonitrile(containing 5 mmol•L-1 ammonium formate and 0.1% formic acid)is mobile phase B.Electrospray ionization(ESI)source was applied by positive ionization in multiple reaction monitoring(MRM)modes. GC-MS/MS analysis was performed on a DM17ms(30 m×0.25 mm, 0.25 μm)capillary column with electron impact(EI)source, electron impact (EI) source was applied by positive ionization in multiple reaction monitoring modes (MRM). RESULTS: The correlation coefficient r of 33 pesticide residues showed good linearity in the linear range of 2 to 20 ng•mL-1 was greater than 0.990 0. The average recoveries at spiked levels of low level and high level (0.01 and 0.04 mg•kg-1), repeat 5 times per level. The average recovery was 87.57%-120.98%, and the RSD was between 1.45%-14.03%. CONCLUSION： The method can quickly and effectively detect pesticide residues in ginseng.
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Objective: This study aimed to simultaneously determine six composition of Panax ginseng by quantitative analysis of multi-components with a single-marker (QAMS) in different paris. Methods: Phenomenex Luna C18 (250 mm × 4.6 mm, 5 μm) was used with mobile phase consisting of acetonitrile-0.1% phosphoric acid for gradient elution at a flow rate of 1.0 mL/min, The column temperature was 25 ℃ and the detection wavelength was 203 nm. Ginsenoside Rb1 was used as reference to establish its relative correction factor of Rg1, Re, Rc, Rb2, Rd, The contents of six components were determined by both external standard method and QAMS. and t test was used to evaluate the feasibility and applicability of QAMS. Results: In a certain linear range, the relative correction factor (RCF) was good, No significant differences were observed between the quantitative results of the two methods. Conclusion: It is feasible and suitable to evaluate the quality of Panax ginseng. It can provide a useful reference to quality control of multi-indexed components in Chinese herbs and traditional Chinese preparations.
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Objective: To explore the solid fermentation process of Panax ginseng by Monascus purpureus, which can transfer some major ginsenoside into rare ginsenoside Rg3 with stronger biological activity. Methods: The static dark culture method was used to perform microbial fermentation; Vanilline-glacial acetic acid method was used to detect the total saponins before and after fermentation, and the ginsenoside Rg3 was detected by HPLC. Results: The optimum process parameters of Monascus purpureus fermentation was fermentation 6 d, fermentation temperature 32 ℃, pH 7.0, and water content of substrate 50%. After 6 d of fermentation, the content of total saponins in fermentation products increased by 40%, and the content of ginsenoside Rg3 was 6.047 mg/g, which was 2.3 times as much as that of non-fermented P. ginseng. According to the change of monomer saponin content along with the fermentation time, it was deduced that the transformation path was Rb1 (Rb2)→Rd→Rh2→Rg3. Conclusion: The solid fermentation process of Monascus purpureus established in this study is reasonable, which not only lays a foundation for the directional production of rare saponins Rg3 but provides a theoretical support for preparing rare ginsenoside in vitro.
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Objective To clone the riboflavin kinase gene of Panax ginseng and perform bioinformatics analysis to construct the prokaryotic expression vector and induce its expression in Escherichia coli. Methods The sequences of comp61599_c0_seq12 were screened from transcriptome database in P. ginseng, and software and online resources were used for its bioinformatics analysis. The 3’ end of sequence was lost and the full-length of 3’ end sequence was obtained by 3’ RACE technology. The complete sequences of cDNA of riboflavin kinase in ginseng was obtained by PCR amplification, and the prokaryotic expression vector of pET-32a-PgRFK was constructed and transformed into E. coli BL-21 for inducing expression. Results A riboflavin kinase gene with length of 1 200 bp was successfully cloned from P. ginseng which encoded 399 amino acid and its prokaryotic expression vector was successfully constructed. The molecular weight of SDS-PAGE electrophoresis results was consistent with prediction. Conclusion The PgRFK gene was successfully cloned and expressed in E. coli.
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Qi-fen fever is a syndrome of Qi-fen syndrome in defensive qi nutrient blood syndrome differentiation. Baihu Decoction is a classic prescription of cooling Qi-fen, but when yang brightness heat at qi aspect is too exuberance to hurt human healthy qi and body fluid, Baihu with Ginseng Decoction will be used for treatment. In order to explore the mechanism of antipyretic effects of Baihu with Ginseng Decoction, this paper applied the network pharmacology method on the basis of summarizing the effects of ginseng on the natural immune system and the relationship between fever and the natural immune system. Firstly, 22 chemically active components of ginseng and their corresponding targets were screened by TCMSP database under limited conditions, and HPO database was used to search for fever-related disease targets. Secondly, the "ginseng-target-fever" network was constructed by STRING database corresponding to ginseng chemical active ingredients target and fever-related disease targets, and Cytoscape software was used to visualize the above network and obtained 18 key nodes according to the relevant characteristic parameters. Finally, The GATHER database platform was applied for the enrichment analysis of the gene function of key nodes. According to the network construction and topological calculation, it suggests that ginseng could activate the natural immune system by regulating MyD88-dependent or MyD88-independent Toll-like receptor signaling pathway to promote the antipyretic effects of Baihu Decoction.
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Objective To investigate the chemical constituents of the rhizomes of Ligusticum chuanxiong and discuss the significance of first discovery of ginsenosides from the plant. Methods The compounds were isolated and repeatedly purified by column chromatographies such as macroperous resin, Sephadex LH-20, silica gel, and preparative TLC as well as semi-preparative RP-HPLC. Their structures were elucidated by physicochemical properties, NMR, and MS spectral analyses. Results Three ginsenoside compounds were isolated from the n-butanol extracts of rhizomes of L. chuanxiong, and their structures were identified as (20S)-ginsenoside Rh1 (1), (20R)-ginsenoside Rh1 (2), and (20R)-ginsenoside Rg3 (3). Conclusion Ginsenosides are isolated from the genus Ligusticum (Umbelliferae) for the first time, it is of great significance for clarifying pharmacodynamic material basis of the rhizomes of L. chuanxiong. These results also provide the reference data for further verifying the relevance of plant evolution and traditional efficacy between L. chuanxiong and Panax ginseng.
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Objective: To obtain the structural characteristics and phylogentic relationships of the chloroplast genome in Araliaceae species. Methods: we used 20 chloroplast genomes which have sequenced as materials, they were from 20 species belonging to 10 genus of Araliaceae. Analysis the differences of genomes and the dilation or shrink of four boundaries for IR, we used MEGA 4.0 to build the phylogenetic tree with Angelica gigas of sibling species as the outgroup and analysis their phylogentic relationships. Results: There was a small difference among the chloroplast genomes size, and the largest difference is 1 909 bp. All of species had existed gene replacements, cemA replaced ycf10, and number of genes existed some differences, they were mainly caused by tRNA. The four boundaries of IR was relatively conservative, only Panax vietnamensi, Panax notoginseng and Schefflera delavayi were special, their boundary genes were in IR. All nodes of the phylogenetic tree of Araliaceae which was based on Angelica gigas were of high supports, and the tree had good resolution to reflect the genetic relationships among Araliaceae. Conclusion: Chloroplast genomes have a lot of information, it can be used to analysis phylogeny among the species which are affinity or faster evolution.
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Objective: In order to explore the anti-oxidant capacity of wild and cultivated ginseng, the anti-oxidase activities and antioxidants contents on ascorbic acid-glutathione (AsA-GSH) cycle were compared. Methods: The activity of superoxide dismutase (SOD) was detected by NBT method and the activity of catalase (CAT) was detected by potassium permanganate titration. The anti-oxidase activities and anti-oxidants contents on AsA-GSH cycle were tested using spectrophotometric determination. The contents of glutamate (Glu), cysteine (Gly), and glycine (Cys) were tested by automatic amino acid analysis. Results: The activities of SOD and CAT in wild ginseng were higher than that of cultivated ginseng. In AsA-GSH cycle, the activities of anti-oxidase such as ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), and glutathione reductase (GR). And the contents of antioxidants such as glutathione (GSH), ascorbic acid (AsA), and dehydroascorbic acid (DHA) were also more in wild ginseng. And then the contents of amino acids (Glu, Gly, and Cys), the precursor of GSH synthesis were high expression in wild ginseng to keep the balance of AsA and GSH recirculation. Conclusion: The anti-oxidase activities and antioxidants content on AsA-GSH cycle in wild ginseng are higher than those in cultivated ginseng, which may be leaded to the antioxidant capacity of wild ginseng is stronger. It will provide a theoretical basis for efficacy differences research on wild and cultivated ginseng.
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Panax ginseng C. A. Mey is one of precious traditional Chinese herbal medicine for two thousand years history, due to its various pharmacological effects and wide utilization in the clinical treatment of tumors and other diseases. Presently tumor has become an important factor threatening human health that the antitumor effect of P. ginseng is attracted great attention. In this paper, the effective components of antitumor action by P. ginseng and its molecular mechanism and structure-activity relationship are summarized. Studies have shown that the main effective components of P. ginseng for antitumor effect are ginsenosides and its metabolic products of intestinal bacteria, ginseng polysaccharides, and ginseng polyacetylenes. The functional mechanism of which are clear relatively now and their main mechanisms including induction of cycle arrest, apoptosis and differentiation of tumor cells, enhancement of immunity to tumor cells, inhibition of tumor cell proliferation, invasion and metastasis, etc. And the molecular mechanism is involved in the regulation of many related genes, proteins, proteases, immune cells, cytokines, and signaling pathways, etc. In addition, the active ingredients of P. ginseng exert antitumor effect in a dose-dependent manner, and the different chemical structures of which lead to different antitumor activity. In conclusion, P. ginseng is abundant with antitumor active ingredients, which is expected to provide safe and effective natural medicine and its preparation for clinical treatment of various tumors in the future.
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OBJECTIVE: To develop a rapid DNA detection kit for DNA extraction and PCR identification of Panax ginseng C. A. Mey. METHODS: The classical DNA extraction and PCR identification methods for Panax ginseng C. A. Mey were modified, and the compositions and reaction conditions of the kit were determined. In addition, the specificity, stability, sensitivity, and repeatability of the kit were evaluated. The genomic DNAs of genuine and counterfeit ginseng goods were extracted by the kit and PCR was performed to identify the authenticity. The purity of the extracted DNA was detected by UV spectrophotometry. Finally, commercially available ginseng samples were verified. RESULTS: The purity of the genomic DNA extracted by the kit was (1.73 ± 0.13) (OD260/OD280), and a fragment between 150 and 200 bp could be amplified only from authentic Panax ginseng C. A. Mey. The specificity of the kit was 100%. The repetitive experiments showed that the average intra-assay CV% and inter-assay CV% of the kit were 2.38% and 2. 62%, respectively. The DNA in solutions diluted by 200 times could still be detected. Stability experiment proved that repeated freeze-thawing for 20 times had no significant effect on the activity of this kit and the test sample could be stored at - 20℃ for one year. The specificity test confirmed that 8 samples among the 10 commercial products were genuine, and 2 were counterfeit. CONCLUSION: The nucleic acid extraction and purity of the DNA detection kit can meet the requirement for identification of Panax ginseng C. A. Mey. The kit has good specificity, high sensitivity, and good stability, so it is suitable for the rapid detection of Panax ginseng C. A. Mey.
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Objective: To construct the expression vector of PgMYB4 gene in Panax ginseng and study its function of the drought resisting in Arabidopsis thaliana. Methods: A P. ginseng gene PgMYB4 was cloned by RT-PCR analysis, further, recombinant plasmid vector with PgMYB4 was transformed into wild-type plants of A. thaliana by Agrobacterium tumefacies-mediated Floral Dip method. Transgenic A. thaliana with the expression of PgMYB4 was obtained, further compared with wild-type plants of A. thaliana, their determination of physiologic index related to drought stress was detected. Results: The cDNA (named a PgMYB4) contained a 735 bp open reading frame, encoded 245 amino acids and the predicted molecular weight was 27.914 KDa; Under the condition of drought stress, the growth of transgenic A. thaliana was obviously better than wild-type A. thaliana. Compared with wild-type A. thaliana, the decreased range of relative chlorophyll content in the leaves of transgenic plants of A. thaliana was smaller and the proline content of transgenic plants of A. thaliana increased significantly. The water loss of transgenic plants of A. thaliana was less than that of Wild-type transgenic plants of A. thaliana. Conclusion: Ginseng PgMYB4 gene has the ability of resistance to drought stress.
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Objective: To establish a new specific and high-throughput method for CYP450 quantification, and to investigate the effect of drugs on rat liver CYP450 using Salvia miltiorrhiza and Panax ginseng as tool drugs. Methods: Mass spectrometry with multiple reaction monitoring (MRM) was applied to quantify nine rat liver enzyme CYP450 isoforms from the rat model with S. miltiorrhiza and P. ginseng. With QconCAT heavy peptides as internal standard, we aimed to assess the effects of S. miltiorrhiza and P. ginseng on CYP450 isoforms. Results: The relative standard deviation and relative error of this method were less than 5.9% and 6.8%, respectively, with good linearity (r2>0.9). Nine CYP450s isoforms (CYP1A1, CYP1A2, CYP2B1, CYP2B2, CYP2C6, CYP2C11, CYP3A1, CYP3A2, and CYP17A1) in rat liver microsome treated with S. miltiorrhiza and P. ginseng were also quantified. Compared with the control, S. miltiorrhiza downregulated the expression levels of CYP1A1, CYP2B2, CYP3A2, CYP2C11, and CYP17A1, while upregulated CYP1A2 and CYP2B1. For the effects of P. ginseng, the expression of CYP1A1 and CYP2B2 was decreased, while CYP1A2, CYP2B1, CYP3A2, CYP2C11, and CYP17A1 were increased. Conclusion: We successfully construct a method with MRM to quantify rat liver CYP450 isoforms. And we firstly quantify CYP2B1, CYP2B2, and CYP17A1 in rat liver. The results reveal the regulation of CYP450 by S. miltiorrhiza and P. ginseng, which could provide clinical application for drug compatibility in practice, and avoid adverse drug reactions.