Panax Japonicus Research Articles

Computational Prediction of Antiangiogenesis Synergistic Mechanisms of Total Saponins of Panax japonicus Against Rheumatoid Arthritis.

Objective: To investigate the anti-angiogenesis mechanisms and key targets of total saponins of Panax japonicus (TSPJ) in the treatment of rheumatoid arthritis (RA). Methods: RStudio3.6.1 software was used to obtain differentially expressed genes (DEGs) by analyzing the differences in gene expression in the synovial tissue of RA and to predict the potential targets of active compounds from TSPJ by the PharmMapper and SwissTargetPrediction databases. We evaluated the overlapping genes by intersectional analysis of DEGs and drug targets. Based on the overlapping genes, we used Cytoscape 3.7.2 software to construct a protein–protein interactions (PPI) network and applied Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis to determine the mechanisms of the treatment. Finally, the correlations with angiogenesis-related genes were explored. Collagen-induced arthritis (CIA) model was established and treated with different doses of TSPJ. The manifestations of CIA were determined by evaluation of arthritis index and histology score. Serum levels of vascular endothelial growth factor (VEGF) and the hypoxia-inducible factor 1 (HIF-1) were tested by ELISA. The mRNA levels of IL-1β and IL-17A were detected by real time-quantitative PCR. Results: Altogether, 2670 DEGs were obtained by differential analysis, and 371 drug targets were predicted for four active components (Araloside A, Chikusetsusaponin IVa, Ginsenoside Rg2, and Ginsenoside Ro). A total of 52 overlapping genes were included in the PPI network and the KEGG analysis. However, only 41 genes in the PPI network had protein interactions. The results of the KEGG enrichment analysis were all related to angiogenesis, including VEGF and HIF-1 signaling pathways. Seven genes with negative correlations and 16 genes with positive correlations were obtained by correlational analysis of DEGs in the VEGF and HIF-1 signaling pathways. SRC proto-oncogene, nonreceptor tyrosine kinase (SRC), and the signal transducer and activator of transcription 3 (STAT 3) had a higher value of degree and showed a significant correlation in the pathways; they were regarded as key targets. Compared with the model group, TSPJ significantly relieved the symptoms and decreased the expression of VEGFA, HIF-1α, IL-1β, and IL-17A in serum or spleens of CIA mice. Conclusion: In the current study, we found that antiangiogenesis is one of the effective strategies of TSPJ against RA; SRC and STAT 3 may be the key targets of TSPJ acting on the VEGF and HIF-1 signaling pathways, which will provide new insight into the treatment of RA by inhibiting inflammation and angiogenesis.

Effect of Panax japonicus saponin Ⅳa on alleviating nonalcoholic steatohepatitis by regulating miR-17-5p/MFN2 signaling pathway

To study the effect of Panax japonicas saponin Ⅳa(SPJ-Ⅳa) on nonalcoholic steatohepatitis(NASH) through miR-17-5 p/MFN2 signaling pathway. The nonalcoholic steatohepatitis model was induced by a high-fat diet combined with CCl_4 in Balb/c male mice. The mouse serum and liver were collected, the body weight and liver weight were measured, the liver index was calculated, and the serum biochemical indicators alanine amino transferase(ALT), triglyceride(TG), and glucose(Glu) were measured. The morphological changes in the liver were detected by HE and Masson staining, Real-time PCR was used to detect lipid metabolism-related genes, inflammation-related genes interleukin-6(IL-6) and interleukin-1β(IL-1β), miR-17-5 p and MFN2 expressions, and Western blot was used to detect MFN2 protein expression level. Compared with the normal control group, the liver index in the HFD+CCl_4 group was significantly increased, and the contents of ALT, TG, and Glu were significantly increased; the morphology showed obvious steatosis and collagen fiber deposition; mRNA expression levels of lipid metabolism-related genes, inflammation-related genes and miR-17-5 p increased significantly, the mRNA expression level of MFN2 decreased significantly, and the protein level of MFN2 decreased. After intervention with SPJ-Ⅳa, the levels of ALT, TG and Glu decreased, morphological steatosis decreased, collagen fiber deposition decreased, and mRNA expression levels of lipid metabolism-related genes, inflammation-related genes and miR-17-5 p decreased. The mRNA expression level of MFN2 increased, and the protein level of MFN2 also increased. The results of this study indicated that miR-17-5 p/MFN2 signaling pathway may be involved in the occurrence and development of NASH, and SPJ-Ⅳa had a protective effect on NASH, its mechanism may be related to the regulation of miR-17-5 p/MFN2 signaling pathway.

Saponins from Panax japonicus ameliorate age-related renal fibrosis by inhibition of inflammation mediated by NF-κB and TGF-β1/Smad signaling and suppression of oxidative stress via activation of Nrf2-ARE signaling

BackgroundThe decreased renal function is known to be associated with biological aging, of which the main pathological features are chronic inflammation and renal interstitial fibrosis. In previous studies, we reported that total saponins from Panax japonicus (SPJs) can availably protect acute myocardial ischemia. We proposed that SPJs might have similar protective effects for aging-associated renal interstitial fibrosis. Thus, in the present study, we evaluated the overall effect of SPJs on renal fibrosis. MethodsSprague-Dawley (SD) aging rats were given SPJs by gavage beginning from 18 months old, at 10 mg/kg/d and 60 mg/kg/d, up to 24 months old. After the experiment, changes in morphology, function and fibrosis of their kidneys were detected. The levels of serum uric acid (UA), β2-microglobulin (β2-MG) and cystatin C (Cys C) were assayed with ELISA kits. The levels of extracellular matrix (ECM), matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), inflammatory factors and changes of oxidative stress parameters were examined. ResultsAfter SPJs treatment, SD rats showed significantly histopathological changes in kidneys accompanied by decreased renal fibrosis and increased renal function; As compared with those in 3-month group, the levels of serum UA, Cys C and β2-MG in 24-month group were significantly increased (p < 0.05). Compared with those in the 24-month group, the levels of serum UA, Cys C and β2-MG in the SPJ-H group were significantly decreased. While ECM was reduced and the levels of MMP-2 and MMP-9 were increased, the levels of TIMP-1, TIMP-2 and transforming growth factor-β1 (TGF-β1)/Smad signaling were decreased; the expression level of phosphorylated nuclear factor kappa-B (NF-κB) was down-regulated with reduced inflammatory factors; meanwhile, the expression of nuclear factor erythroid 2-related factor 2-antioxidant response element (Nrf2-ARE) signaling was aggrandized. ConclusionThese results suggest that SPJs treatment can improve age-associated renal fibrosis by inhibiting TGF-β1/Smad, NFκB signaling pathways and activating Nrf2-ARE signaling pathways and that SPJs can be a potentially valuable anti-renal fibrosis drug.

Synthesis of 3α,20S-Dihydroxydammar-24-en-12-One β-D-Glucopyranosides

3α,20S-Dihydroxydammar-24-en-12-one 3-, 20-, and 3,20-di-O-β-D-glucopyranosides, close structural analogs of chikusetsusaponin-LT8 glycoside from Panax japonicus, were synthesized for the first time. Condensation of 3,20S-dihydroxydammar-24-en-12-one (1) with 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide (6) in the presence of Ag2O gave a mixture of the three acetylated 3-, 20-, and 3,20-di-O-β-Dglucopyranosides 7–9 with 7 dominating (34.6%). Glycosylation of 3α-acetoxy-20S-hydroxydammar-24-en-12-one by glycosyl donor 6 led to regio- and stereoselective formation of acetylated 20-O-β-Dglucopyranoside 10 (48.6%). Subsequent removal of the protecting groups by sodium methoxide gave the corresponding free 3-, 20-, and 3,20-di-O-β-D-glucopyranosides 11–13.

Ecological stoichiometric characteristics of soil and microbial biomass C, N, P in rhizosphere soil of Paris polyphylla and Panax japonicus cultivated understory

Comparison of total organic carbon(TOC), total nitrogen(TN), total phosphorus(TP), soil microbial biomass carbon(MBC), nitrogen(MBN), phosphorus(MBP) and their stoichiometric ratios measuring from understory planting of Paris polyphylla and Panax japonicus rhizosphere soil with the data of the original forest soil will help us to understand the influence of different planting patterns between soil traits and soil microbial interaction and nutrient cycle characteristics. The results showed that the contents of TOC, TN and MBN were the highest in the rhizosphere soil of P. japonicus, while the highest values of TP, MBC and MBP were found in the rhizosphere soil of P. polyphylla. The changes of TOC∶TN, TOC∶TP, TN∶TP, MBC∶MBN, MBC∶MBP and MBN∶MBP of P. polyphylla and P. japonicus rhizosphere soil compared with the data of the original forest soil were 3.65 and 37.32%,-14.89 and 82.23%,-17.87 and 32.76%, 25.67 and-50.60%,-75.95 and-16.33% as well as-80.79 and 69.76%, respectively. TN and TP were significantly correlated with MBN and MBP respectively. Although, monoculture of P. polyphylla and P. japonicus changed soil nutrient level, it did not reach the state of nutrient deficiency. The demands for nitrogen and phosphorus between P. polyphylla and P. japonicus were quite different. According to their different habits, monoculture of P. polyphylla and P. japonicus could change the understory soil traits, resulting in allometric changes in part of soil nutrient stoichiometry and soil microbial stoichiometry, and then the disappearance of internal stability. The variations in the understory soil caused by P. polyphylla and P. japonicus is developing in completely different directions, whether this phenomenon indicates that the two species have less niche overlap needs further study.

A mathematical model for quality evaluation of total saponins of Panax japonicas based on hypolipidemic activity

ObjectiveThe chemical finger printing-based methods for evaluating TCMs quality can report partial of TCMs quality without linking to effective constituents. In this study, a mathematical model was established for the quality evaluation of total saponins of Panax japonicus (TSPJ), a folk medicine in China and Japan for treating diseases, through coupling the dynamic changes of chemical constitutions with corresponding activities. MethodsHigh-performance liquid chromatography (HPLC) fingerprints were applied to establish the chromatographic database of TSPJ. The associated hypolipidemic activity database was determined by TG assay using HepG2 cell model. Correlation analyses of two databases were performed by partial least squares (PLS) for calculating regression coefficients, and the interval value of YZL value (the ratio of positive and negative peak-to-peak area coefficient) closely related to hypolipidemic activity was refined by the formula of Norminv function to value the quality of TSPJ. ResultsIn this study, the chromatographic data of 16 common peaks were obtained from 20 batches of TSPJ. After the estimate by this mathematical evaluation model, seven peaks were positively correlated with hypolipidemic activity, and nine peaks were negatively correlated with hypolipidemic activity. When the YZL value was less than 0.7861, the quality of sample was inferior, while YZL value was more than 6.6992, and the quality of samples was superior. The quality of another ten batches of TSPJ was further assessed to verify this method. ConclusionThese results indicated that the established model could be usefully applied to evaluate the quality of TSPJ in the hypolipidemic activity.

The Preventive Effect of Total Saponins from &lt;i&gt;Panax japonicas&lt;/i&gt; on Inflammation and Insulin Resistance in Adipose Tissue of Mice Induced by High-Fat Diet

Background: In the present research, we aimed to investigate the effects of total saponins from Panax japonica (TSPJ) on adipocyte inflammation and insulin resistance induced by a high-fat diet (HFD). Moreover, its underlying mechanism was also further explored. Methods: C57/BL mice were randomly divided into four groups as follows: normal diet (ND) group, HFD group, and two TSPJ + HFD groups (30 mg/kg and 90 mg/kg). Insulin resistance test, glucose tolerance test, fat index analysis, histopathological assay, Western blotting analysis, and real-time PCR were performed to analyze the protective effect of TSPJ on adipose tissue. The protective mechanism of TSPJ on 3T3-L1 cells was studied in vitro by Oil Red O staining, 2-deoxyglucose uptake measurement etc. . Findings: In vivo, we found that TSPJ inhibited the activation of NF-κB and increase the transcription of p-AKT, leading to reduced inflammation and improved the sensitivity of adipocytes to insulin. In vitro evaluation revealed that palmitic acid (PA) induced inflammation by activating the NF-κB pathway, and induced insulin resistance by affecting the expressions of p-AKT in adipocytes. TSPJ pre-treatment could prevent such these response in a dose-dependent manner. TSPJ might exert its protective effect on 3T3-L1 cells by affecting the expression of AMPK. Interpretations: Our study showed that TSPJ could inhibit adipocyte inflammation by inhibiting NF-κB pathways. In addition, TSPJ could also affect the expressions of p-AKT to increase insulin sensitivity in adipocytes via targeting AMPK. Funding: This study was financially supported by the grants from National Natural Science Foundation of China. Declaration of Interests: The authors declare that they have no competing interests. Ethics Approval Statement: This study was financially supported by the grants from National Natural Science Foundation of China (Grant No. 81773959 to C.F. Yuan and No. 81974528 to C.F. Yuan) and Health commission of Hubei Province scientific research project in China. (Grant No. WJ2019H527 to C.F. Yuan).

Antihypoxic Action of Panax Japonicus, Tribulus Terrestris and Dioscorea Deltoidea Cell Cultures: In Silico and Animal Studies.

Chemical diversity of secondary metabolites provides a considerable variety of pharmacological actions with a significant extension due to their combinations in plant extracts. Production of plant-derived medicinal products in cell cultures has advantages because of the efficient use of different biotic and abiotic elicitors and better control of the developmental processes. Using PASS software, we predicted biological activity spectra for phytoconstituents identified in cell cultures of Panax japonicus (12 molecules), Tribulus terrestris (4 molecules), and Dioscorea deltoidea (3 molecules). Mechanisms of action associated with the antihypoxic effect were predicted for the majority of molecules. PharmaExpert software allowed analyzing possible synergistic or additive effects of the combinations of phytoconstituents associated with the antihypoxic action. Experimental studies of the antihypoxic effect of the plants' extracts in water and ethanol have been performed in 3 animal models: Acute asphyctic hypoxia (AAH), Acute haemic hypoxia (AHeH), and Acute histotoxic hypoxia (AHtH). Effects of Panax japonicus and Tribulus terrestris preparations exceeded the activity of the reference drug Mexidol in the AHtH model. In the AHeH model, all preparations demonstrated moderate activity; the most potent has been observed for Dioscorea deltoidea. Thus, we found that experimental studies in animal models have confirmed the in silico prediction.

Full-length transcriptome sequencing and modular organization analysis of oleanolic acid- and dammarane-type saponins related gene expression patterns in Panax japonicus

The saponins found in Panax japonicus, a traditional medicinal herb in Asia, exhibit high degrees of structural and functional similarity. In this study, metabolite analysis revealed that oleanolic acid-type and dammarane-type saponins were distributed unevenly in three tissues (rhizome_Y, rhizome_O, and secRoot) of P. japonicus. Single-molecule real-time (SMRT) sequencing and next generation sequencing (NGS) data revealed distinct and tissue-specific transcriptomic patterns relating to the production of these two types of saponins. In the co-expression network and hierarchical clustering analyses, one 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) and two 1-deoxy-D-xylulose-5-phosphate synthase (DXS) etc. transcripts were found to be key genes associated with the biosynthesis of oleanolic acid and dammarane-type saponins in P. japonicus, respectively. In addition, cytochrome p450 (CYP) and UDP-glucuronosyltransferase (UGT) family proteins that serve as regulators of saponin biosynthesis-related genes were also found to exhibit tissue-specific expression patterns. Together these results offer a comprehensive metabolomic and transcriptomic overview of P. japonicus.

Saponins of Panax japonicus Confer Neuroprotection against Brain Aging through Mitochondrial Related Oxidative Stress and Autophagy in Rats.

Oxidative stress and mitochondrial dysfunction play a vital role in the pathogenesis of brain aging. Saponins from Panax japonicus (SPJ) have attracted much attention for their potential to attenuate age-related oxidative stress as the main ingredient in rhizomes of Panax japonicus. This study aimed to investigate the neuroprotective effects of SPJ on natural aging rats as well as the underlying mechanisms regarding oxidative stress and mitochondrial pathway. Sprague-Dawley rats were divided into control groups (3-, 9-, 15- and 24-month old groups) and SPJ-treated groups. For SPJ-treated groups, SPJ were orally administrated to 18-month old rats at doses of 10 mg/kg, 30 mg/kg and 60 mg/kg once daily. Control groups were given the same volume of saline. After the treatment with SPJ or saline for six months, the cortex and hippocampus were rapidly harvested and deposited at -80°C after the rats were decapitated under anesthesia. The neuroprotective effects of SPJ were estimated by histopathological observation, TUNEL detection, biochemical determination and western blotting. SPJ improved pathomorphological changes in neuronal cells and decreased apoptosis in the cortex and hippocampus of aging rats, increased the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), Na+/K+-ATPase, Ca2+-ATPase and Ca2+/Mg2+-ATPase whereas, decreased malondialdehyde (MDA) contents in the cortex of aging rats. Furthermore, the SPJ increased silent mating type information regulation 2 homolog-1 (SIRT1) protein expression, decreased acetylated level of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) in the cortex and hippocampus of aging rats, and reversed the aging-induced decline of Forkhead box O3 (Foxo3a), Superoxide Dismutase 2 (SOD2), microtubule-associated protein light chain 3 (LC3II) and Beclin1 levels in the cortex and hippocampus. Our data showed that SPJ conferred neuroprotection partly through the regulation of oxidative stress and mitochondria-related pathways in aging rats.

Anticoagulant active ingredients identification of total saponin extraction of different Panax medicinal plants based on grey relational analysis combined with UPLC-MS and molecular docking

Ethnopharmacological relevanceIn traditional use, Panax medicinal plants (ginseng, red ginseng, notoginseng, Panax japonicus, and Panacis majoris rhizoma) have different bioactivities from each other, even under different dosages, but their chemical compositions are very similar; so the question is, what is the primary effective substance induced the different efficacy, and how to identify them from a group of chemical constituents? Aim of the studyThe goal of this research was to provide a strategy to determine the effective substance in Panax genus medicinal plants responsible for the anticoagulant response. Materials and methodsThis research used ultra-high performance liquid chromatography coupled with mass spectrometry (UPLC-MS) to analyze the saponin chemical compositions of different concentration ethanol extraction of notoginseng and the ginseng genus medical plant 75% ethanol extraction (Panax ginseng, red ginseng, Panax japonicus, Panacis majoris rhizome), and used four coagulation factors (PT, TT, APTT, Fib) to evaluate the anticoagulant activity of the extracts. Grey correlation analysis was applied to establish the spectral effect relationship and give the anticoagulant potency of different saponins. Network pharmacology and molecular docking were adopted to clarify and verify the possible mechanisms of anticoagulant action. ResultsThe results showed that the blood physiological regulation activities of Panax medicinal plants were different according to the solvent concentration, processing, species and dosage. Overall, the most suitable solvent for extraction of SQ was 75% ethanol; At low dosage (10–100 mg/mL), the anticoagulant effect of Panax medical plants was: ZJS > ZZS > SQ > RS > HS, and at high doses (100–1000 mg/mL) was: SQ > ZJS > ZZS > RS > HS. GRA and molecular docking results showed the contribution of some components (NG-R2, NG-Fc/G-Ra1/G-Ra2, G-Rc, G-Rk3, and G-Rh4) to the whole anticoagulant activity of the drug were increased, while the effect of CS-IVa was just decreased with the increase of dosage; the anticoagulant effect of G-Rg3 (the main anticoagulant component) is mainly related to the targets F2, AR, RHO, ACR, MB, GZMB, B2M, CA2, CAT, and PAPOLA. ConclusionThis study determined the effective substance of anti-coagulation of ginseng genus herbal medicines and the regulation of different anticoagulant effects of TCM by changing various influencing conditions, including processing method, extraction method, and dose. It also provided an effective strategy for effective substances identification of multicomponent, multifunction, and multipurpose herbal medicine.

Complete chloroplast genome of Panax japonicus var. major (Burkill) C. Y. Wu & K. M. Feng (Araliaceae) and phylogenetic analysis

Panax japonicus var. major (Araliaceae) is an important medicinal plant, whose rhizomes have pharmacological activities such as protection of cardiovascular and cerebrovascular system, central nervous system and so on. In this study, we sequenced complete chloroplast (cp) genome sequence of P. japonicus var. major so as to explore phylogenetic relationship between this species and related species. The results showed that complete cp genome of P. japonicus var. major was 156,402 bp in length, consisting of a large single copy (LSC) region of 86,187 bp, a small single copy (SSC) region of 18,007 bp, and two inverted repeat regions (IRa and IRb) of 26,104 bp. In total, 114 genes were annotated, comprising of 80 protein-coding genes, 30 tRNA genes, and four rRNA genes. The phylogenetic analysis indicated that P. japonicus var. major was closely related to its original variety, P. japonicus var. japonicus; meanwhile, Panax was a monophyletic group with high support value and it had a close relationship with Aralia in Araliaceae.

Inhibitory Effects of Essential Oil Extracts From Panax Plants Against β-Secretase, Cholinesterase, and Amyloid Aggregation

Numerous pharmacological studies on Panax plants have been performed. However, these studies were limited to ginsenosides, which are typical constituents in Panax plants. In our research program to discover novel agents to prevent dementia and improve dementia symptoms, especially Alzheimer’s disease (AD), we investigated the inhibitory activities of essential oil (EO) extracts from 4 Panax plants against β-secretase, amyloid β (Aβ) aggregation, acetylcholinesterase (AChE), and butyrylcholinesterase (BChE). An EO extract of Panax japonicus showed the most potent activity with 51.3% inhibition at 500 μg/mL against β-secretase. Panax ginseng showed the most potent inhibitory activity against AChE and BChE with 70.4% and 84.4% inhibition at 50 μg/mL, respectively. Panax notoginseng extract showed the most potent activity with 57.3% inhibition at 500 μg/mL against Aβ aggregation. From these results, an EO extract of P. ginseng could be an effective agent to improve AD symptoms, while EO extracts of P. japonicus and P. notoginseng could be suitable for AD prevention.

A novel preparation of Panax japonicas with high bioequivalence of ginsenoside Rg3

A novel preparation of Panax japonicas with high bioequivalence of ginsenoside Rg3

Comparative transcriptome analysis reveals a gene expression profile that contributes to rhizome swelling in Panax japonicus var. major

Panax japonicus var. major is a rare and endangered Chinese traditional medicinal plant that simultaneously forms two types of rhizomes (Swp: Swollen part and Slp: Slender part). However, the mechanism controlling rhizome enlargement in P. japonicus var. major is poorly understood. In this study, the mechanism of rhizome enlargement was investigated by comparing the differences of internal anatomy, hormone contents, starch content and molecular transcription levels between Swp and Slp of in P. japonicus var. major. Illumina sequencing generated 1,333 differentially expressed genes (DEGs) that were differentially expressed between the two samples. Pathway enrichment analysis revealed that DEGs involved in starch and sucrose metabolism, diterpenoid biosynthesis and plant hormone signal transduction pathway were significantly enriched. Additionally, starch determination results showed that starch content in the swollen part was significantly higher than that in the slender part. Furthermore, hormone determination results showed that gibberellin A1 (GA1), gibberellin A53 (GA53), indole-3-acetic acid (IAA) and jasmonic acid (JA) contents were negatively correlated, while cis-zeatin (cZ) content was positively correlated with rhizome enlargement. The results of this research screened out several DEGs and potential candidate genes for rhizome enlargement, which lays the foundation for the identification of expanded genes in Panax species.

Synthesis of 12‐epi‐Protopanaxadiol and Formal Synthesis of Ginsenoside Chikusetsusaponin‐LT8

In the context of the total synthesis of protopanaxadiol, two strategies were explored. One strategy from an optically active trienic epoxide, possessing a 5‐membered ring, prepared from (S)‐epoxy‐limonene and (S)‐epoxyfarnesol, which was submitted to Ti‐(III)‐mediated radical cascade to afford an original tetracyclic structure resulting from a 6‐endo‐trig 6‐endo‐trig 8‐endo‐trig process. A second strategy, starting from a Wieland‐Miescher‐type ketone, using a “ring‐by‐ring” synthesis allowed the synthesis of 12‐epi‐protopanaxadiol. In this latter strategy, an efficient sequence of reactions to install the two vicinal C8–C14 quaternary centers involves: i) a Barbier type reaction; ii) an oxidative allylic transposition and iii) a nickel‐catalyzed addition of trimethylaluminium. By using this second strategy, 20‐hydroxydammar‐24‐ene‐3,12‐dione was synthesized which represents a formal synthesis of chikusetsusaponin‐LT8, isolated from Panax japonicus.

Integration of Data-Dependent Acquisition (DDA) and Data-Independent High-Definition MSE (HDMSE) for the Comprehensive Profiling and Characterization of Multicomponents from Panax japonicus by UHPLC/IM-QTOF-MS.

The complexity of herbal matrix necessitates the development of powerful analytical strategies to enable comprehensive multicomponent characterization. In this work, targeting the multicomponents from Panax japonicus C.A. Meyer, both data dependent acquisition (DDA) and data-independent high-definition MSE (HDMSE) in the negative electrospray ionization mode were used to extend the coverage of untargeted metabolites characterization by ultra-high-performance liquid chromatography (UHPLC) coupled to a VionTM IM-QTOF (ion-mobility/quadrupole time-of-flight) high-resolution mass spectrometer. Efficient chromatographic separation was achieved by using a BEH Shield RP18 column. Optimized mass-dependent ramp collision energy of DDA enabled more balanced MS/MS fragmentation for mono- to penta-glycosidic ginsenosides. An in-house ginsenoside database containing 504 known ginsenosides and 60 reference compounds was established and incorporated into UNIFITM, by which efficient and automated peak annotation was accomplished. By streamlined data processing workflows, we could identify or tentatively characterize 178 saponins from P. japonicus, of which 75 may have not been isolated from the Panax genus. Amongst them, 168 ginsenosides were characterized based on the DDA data, while 10 ones were newly identified from the HDMSE data, which indicated their complementary role. Conclusively, the in-depth deconvolution and characterization of multicomponents from P. japonicus were achieved, and the approaches we developed can be an example for comprehensive chemical basis elucidation of traditional Chinese medicine (TCM).

Phylogenetic relationship and characterization of the complete chloroplast genome of Panax notoginseng, the endemic medicinal herbs to China

Panax notoginseng is the most important valued and endemic medicinal herb to China. It belongs to the Araliaceae family, which has a longer medical history in China. The complete chloroplast genome of Panax notoginseng is 156,387 bp in size and displays a typical quadripartite structure of the large single-copy region (LSC, 86,112 bp), small single-copy region (SSC, 18,005 bp) that separate by a pair of inverted repeat regions (IRs, each for 26,135 bp). The base nucleotide composition of the cpDNA is 30.8% of A, 31.1% of T, 19.9% of C, and 18.2% of G, with a total G + C content of 38.1%. The whole chloroplast genome of P. notoginseng contains 134 genes, including 89 protein-coding genes (PCGs), 37 transfer RNA (tRNAs) genes, and eight ribosomal RNA (rRNAs) genes species. Phylogenetic relationship analysis based on 37 medicinal herbs species confirmed the position of P. notoginseng closely related to Panax japonicus and Panax vietnamensis.

Triterpenoid Saponins from Tu Jia Ethnomedicine Bai San Qi and Their Cytotoxicity on Hep G2 and BGC-823 Cell Lines

ObjectiveTu Jia ethnomedicine is a unique medical system inherited and adhibited by Tu Jia minority living in central-south China. Panax japonicus C. A. Mey. (Bai San Qi, 白三七) is recognized as an effective and rare medicinal plant to treat weakness, fatigue and rheumatism in Tu Jia ethnomedicine. This paper is to discover more substance evidence for the application of Tu Jia ethnomedicine. MethodsColumn chromatography and preparative high performance liquid chromatography (HPLC) was applied for isolation and purification; 1H-NMR, 13C-NMR, 1H-1H COSY, HSQC and HMBC NMR spectra were applied for structure identification; Methyl thiazolyl tetrazolim (MTT) assays were applied for cytotoxicity evaluation. ResultsTotally 12 known compounds were isolated by column chromatography and preparative HPLC from rhizomes of Panax japonicus C. A. Mey. (Bai San Qi, 白三七). Structures of these compounds were identified by their NMR spectra. All the 12 compounds were triterpenoid saponins. Five of them were oleanolic acid type, and the remaining 7 were dammarane type. Eleven compounds were assayed for their cytotoxic activity against Hep G2 human liver cancer cell lines and BGC-823 human gastric cancer cell lines. Three of the 11 showed relatively dominant cytotoxicity against these cell lines. ConclusionsA total of 12 known compounds have been identified from Panax japonicus C. A. Mey. (Bai San Qi, 白三七); NMR spectra of compounds with similar skeletons showed regular characteristics; 3 compounds showed relatively dominant cytotoxicity against Hep G2 and BGC-823 cancer cell lines, and the result can be valued as weak while setting the taxol as a positive control.

Analysis of gene expression of Tetrahymena thermophila treated with Panax japonicas

Panax japonicus is a traditional Chinese medicine,and its principle components have shown certain pharmacological activities for cell damage,aging and cell apoptosis. In order to clarify the pharmacological mechanism and involved metabolic pathways of P. japonicas,the gene expression of Tetrahymena thermophila under P. japonicus treatment was analyzed through high-throughput transcriptome sequencing in this study. Based on the transcriptome analysis,3 544 differentially expressed genes were identified in control group,of which 1 945 genes showed up-regulated expression and 1 599 genes showed down-regulated expression. Under P. japonicas treatment in the experiment group,3 312 differentially expressed genes were screened,of which 1 `493 genes showed up-regulated expression and 1 819 genes showed down-regulated expression. GO enrichment analysis indicated that in control group,the genes in the cells in a series of fundamental biological process were down-regulated,such as DNA replication and protein synthesis; while the signal transduction process and fatty acids oxidizing process were enriched. Whereas in the experiment group,down-regulated genes were mainly enriched in oxidation-reduction,cofactor metabolic process and vitamin metabolic process; up-regulated genes were enriched in signal transduction process and protein modification process. In the analysis using KEGG database,cell cycle pathway was enhanced and autophagy pathway was inhibited under the condition of P. japonicas treatment. Real-time quantitative polymerase chain reaction( RT-qPCR) was used to detect the expression differences between 6 up-regulated and 4 down-regulated genes in related metabolic pathways. The RT-q PCR results and RNA-Seq data were highly correlated and consistent with each other. This study could provide important direction and basis for further study on the mechanism of cell growth regulation with the treatment of P. japonica.