Total Ginsenosides Research Articles

Functional regulation of ginsenoside biosynthesis by RNA interferences of a UDP-glycosyltransferase gene in Panax ginseng and Panax quinquefolius

Panax ginseng (Asian ginseng) and Panax quinquefolius (American ginseng) have been used as medicinal and functional herbal remedies worldwide. Different properties of P. ginseng and P. quinquefolius were confirmed not only in clinical findings, but also at cellular and molecular levels. The major pharmacological ingredients of P. ginseng and P. quinquefolius are the triterpene saponins known as ginsenosides. The P. ginseng roots contain a higher ratio of ginsenoside Rg1:Rb1 than that in P. quinquefolius. In ginseng plants, various ginsenosides are synthesized via three key reactions: cyclization, hydroxylation and glycosylation. To date, several genes including dammarenediol synthase (DS), protopanaxadiol synthase and protopanaxatriol synthase have been isolated in P. ginseng and P. quinquefolius. Although some glycosyltransferase genes have been isolated and identified association with ginsenoside synthesis in P. ginseng, little is known about the glycosylation mechanism in P. quinquefolius. In this paper, we cloned and identified a UDP-glycosyltransferase gene named Pq3-O-UGT2 from P. quinquefolius (GenBank accession No. KR106207). In vitro enzymatic activity experiments biochemically confirmed that Pq3-O-UGT2 catalyzed the glycosylation of Rh2 and F2 to produce Rg3 and Rd, and the chemical structure of the products were confirmed susing high performance liquid chromatography electrospray ionization mass spectrometry (HPLC/ESI-MS). High sequence similarity between Pq3-O-UGT2 and PgUGT94Q2 indicated a close evolutionary relationship between P. ginseng and P. quinquefolius. Moreover, we established both P. ginseng and P. quinquefolius RNAi transgenic roots lines. RNA interference of Pq3-O-UGT2 and PgUGT94Q2 led to reduce levels of ginsenoside Rd, protopanaxadiol-type and total ginsenosides. Expression of key genes including protopanaxadiol and protopanaxatriol synthases was up-regulated in RNAi lines, while expression of dammarenediol synthase gene was not obviously increased. These results revealed that P. quinquefolius was more sensitive to the RNAi of Pq3-O-UGT2 and PgUGT94Q2 when compared with P. ginseng.

Pathogenic fungal elicitors enhance ginsenoside biosynthesis of adventitious roots in Panax quinquefolius during bioreactor culture

Fungi have been widely used as biotic elicitors to promote metabolite production of plant cell, tissue, and organ cultures. Alternaria panax Whetz. and Cylindrocarpon destructans (Zinss) Scholten are the main pathogenic fungi in ginseng cultivation. The present study used extracts of A. panax and C. destructans as elicitors to simulate adventitious roots (ARs) of Panax quinquefolius L. for improvement of ginsenoside accumulation during AR bioreactor culture. To select a suitable elicitation method, the effects of treatment duration and concentration of both fungal extracts were investigated. After 30 d of AR bioreactor culture, the addition of A. panax and C. destructans extracts for 8 d to the culture medium produced higher ginsenoside content and productivity than the other treatment periods. During the elicitation of both fungal extracts, the peak contents of nitric oxide (NO), putrescine (Put), and ginsenoside were observed at 2, 4, and 8 d, respectively. Therefore, NO and Put could be upstream signals to regulate ginsenoside synthesis. The optimal elicitor concentration varied between extracts of A. panax and C. destructans. The maximum total ginsenoside content and productivity were observed with 4mg/L and 20mg/L of the A. panax and C. destructans extracts, respectively. The ginsenoside monomers with the most increase were Rg3, Rh2, and Re for the A. panax extract and Rg3, Rh2, and Rf for the C. destructans extract. In addition, the AR biomass was more inhibited by the C. destructans (20mg/L) extract, although its ginsenoside content (34.1mg/g DW) was higher than that with the A. panax (4mg/L) extract. The maximum ginsenoside production (276.0mg/L) was achieved with the A. panax (4mg/L) extract. Therefore, the A. panax extract was more suitable as an elicitor for ginsenoside production during AR bioreactor culture. The present study confirmed that treating 30-d-old P. quinquefolius ARs with 4mg/L A. panax extract for 8 d during bioreactor culture is the optimal elicitation method; these ARs can be applied to the commercial production of P. quinquefolius products.

Effect of Controlled Light Environment on the Growth and Ginsenoside Content of Panax ginseng C. A. Meyer

Background: The photosynthetic efficiency cool-season, semi-shade ginseng is normal at low morning temperatures, but drops at high afternoon temperatures. Therefore, optimal plant performance would be ensured if it were possible to control daily light transmission rates (LTR). Methods and Results: Plants were grown in a controlled light environment that replicated 11 AM conditions and comparatively analyzed against plant grown under normal conditions. Growth in the controlled light environment resulted in a 2.81 fold increase in photosynthetic efficiency with no change in chlorophyll content, although LTR were high due to low morning temperatures. Increased aerial plant growth was observed in the ginseng plants adapted to the controlled light environment, which in turn influenced root weight. An 81% increase in fresh root weight (33.3 g per plant on average) was observed in 4-year-old ginseng plants grown in controlled light environment compared to the plants grown following conventional practices (18.4 g per plant on average). With regard to the inorganic composition of leaves of 4-year-old ginseng plants grown in controlled light environment, an increased in Fe content was observed, while Mn and Zn content decreased, and total ginsenoside content of roots increased 2.37 fold. Conclusions: Growth of ginseng under a favorable light environment, such as the condition which exist naturally at 11 AM and are suitable for the plant's photosynthetic activity creates the possibility of large scale production, excellent-quality ginseng.

Chemical and bioactive comparison of flowers of Panax ginseng Meyer, Panax quinquefolius L., and Panax notoginseng Burk.

BackgroundAlthough flowers of Panax ginseng Meyer (FPG), Panax quinquefolius L. (FPQ), and Panax notoginseng Burk. (FPN) have been historically used as both medicine and food, each is used differently in practice. MethodsTo investigate the connection between components and enhancing immunity activity of FPG, FPQ, and FPN, a method based on a rapid LC coupled with quadrupole time-of-flight MS and immunomodulatory activity study evaluated by a carbon clearance test were combined. ResultsAccording to quantitative results, the ratio of the total content of protopanaxatiol-type ginsenosides to protopanaxadiol-type ginsenosides in FPN was 0, but ranged from 1.10 to 1.32 and from 0.23 to 0.35 in FPG and FPQ, respectively. The ratio of the total content of neutral ginsenosides to the corresponding malonyl-ginsenosides in FPN (5.52±1.33%) was higher than FPG (3.2±0.64%) and FPQ (2.39±0.57%). The colorimetric analysis showed the content of total ginsenosides in FPQ, FPG, and FPN to be 13.75±0.60%, 17.45±0.42%, and 12.45±1.77%, respectively. The carbon clearance assay indicated that the phagocytic activity of FPG and FPQ was higher than that of FPN. A clear discrimination among FPG, FPQ, and FPN was observed in the principal component analysis score plots. Seven compounds were confirmed to contribute strongly by loading plots, which may be the cause of differences in efficacy. ConclusionThis study provides basic information about the chemical and bioactive comparison of FPG, FPQ, and FPN, indicating that protopanaxtriol-type ginsenosides and malonyl-ginsenosides may play a key role in their enhancing immunity properties.

Study on relationship between effective components and soil enzyme activity in different growth patterns of Panax ginseng

Study on 5 effective components and 6 soil enzyme activities of 2 different growth patterns, analyse the dates with the canonical correlation analysis, In order to reveal the relations between the effective components and soil enzyme activities. The result showed that they had a great relation between the effective components and soil enzyme activities, the activity of the same enzyme in humus soil was higher than that in farmland soil. Growth pattern of farmland soil, if the invertase and phosphatase activity were too high, which would inhibit the accumulation of total ginsenoside, water-miscible total proteins and total amino acid; Growth pattern of humus soil, if the invertase, urease and phosphatase activity were too high, which would inhibit the accumulation of total ginsenoside and the total essential oils. Integral soil enzyme activity can be used as a index of soil quality, which, together with other growth factors. The appropriate enzyme activity can accelerate the circulation and transformation of all kinds of material in the soil, improve effectively components accumulation.

Antimelanogenesis Activity of Hydrolyzed Ginseng Extract (GINST) via Inhibition of JNK Mitogen-activated Protein Kinase in B16F10 Cells.

GINST is a hydrolyzed ginseng extract produced by an in vitro process that imitates the metabolic function of bacteria in the human digestive track and has approved by the Ministry of Food and Drug Safety of Korea for the management of postprandial hyperglycemia. Additionally, GINST has been reported to have other physiological functions including anti-aging and antioxidant effects. The objectives of this study are to compare the antimelanogenic effects of fresh ginseng extract (FGE) and GINST extract and to elucidate the functional mechanism. The concentration of total ginsenosides in FGE and GINST was measured using ultraperformance liquid chromatography with a C18 column. B16F10 cells were treated with FGE and GINST for 72 h to assess melanin content, tyrosinase activity, and protein levels of microphthalmia-associated transcription factor (MITF) and tyrosinase-related protein-1 (TRP-1). The activity of kinases involved in mitogen-activated protein kinase (MAPK) signaling, such as extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinases (JNK), and p38 mitogen-activated protein kinases (p38), were measured using western blots. While neither FGE nor GINST inhibited the activity of mushroom tyrosinase directly, GINST decreased melanogenesis and tyrosinase activity markedly. Furthermore, our results indicate that GINST downregulated the levels of MITF and TRP-1 possibly by suppressing JNK signaling. We concluded that, when compared to FGE, GINST has a superior antimelanogenic effect mediated by the downregulation of MITF, TRP-1, and intracellular tyrosinase activity via the JNK signaling pathway. Thus, we suggest that GINST has the potential to be used as a novel skin whitening agent.

Variation of Ginsenosides in Ginseng of Different Ages

Panax ginseng has been used in traditional oriental medicine for thousands of years. Ginsenosides, the major chemical components of the roots, are considered to be responsible for the medicinal properties of ginseng. Ginsenosides increase with the age of ginseng root in general knowledge, and in this study the content of ginsenosides in ginseng of different ages was quantified. Separation and determination of eight main ginsenosides, Rg1, Re, Rb1, Rc, Rg2, Rb2, Rb3 and Rd, was performed by high performance liquid chromatography with UV detection at 203 nm. The content of Rg1, Re, Rb1, Rc, Rg2 and Rd increased from 5 to 16-year-old ginseng and then decreased, while Rb2 and Rb3 increased in the range of 5-12 years, but then slowly decreased. However, the total eight ginsenosides in 16 year old ginseng had a higher content than that in any other from 5-18 years old. As a result, the content of ginsenosides and total ginsenosides was not positively related to age from 5-18 years, which is not in full agreement with the general knowledge of ginseng. Thus, this study suggests that the older wild ginseng may not result in better medicinal ginseng for herbal medicines.

Metabonomics Approach to Assessing the Metabolism Variation and Endoexogenous Metabolic Interaction of Ginsenosides in Cold Stress Rats.

Metabolic profiling technology, a massive information provider, has promoted the understanding of the metabolism of multicomponent medicines and its interactions with endogenous metabolites, which was previously a challenge in clarification. In this study, an untargeted GC/MS-based approach was employed to investigate the urinary metabolite profile in rats with oral administration of ginsenosides and the control group. Significant changes of urinary metabolites contents were observed in the total ginsenosides group, revealing the impact of ginsenosides as indicated by the up- or down-regulation of several pathways involving neurotransmitter-related metabolites, tricarboxylic acid (TCA) cycle, fatty acids β-oxidation, and intestinal microflora metabolites. Meanwhile, a targeted UPLC-QQQ/MS-based metabonomic approach was developed to investigate the changes of urinary ginsenoside metabolites during the process of acute cold stress. Metabolic analysis indicated that upstream ginsenosides (rg1, re, and rf) increased significantly, whereas downstream ginsenosides (ck, ppd, and ppt) decreased correspondingly after cold exposure. Finally, the relationships between ginsenosides and significantly changed metabolites were investigated by correlation analysis.

Suppression of Hepatic Cyp1a2 by Total Ginsenosides in Lipopolysaccharide-Treated Mice and Primary Mouse Hepatocytes.

The roots of Panax ginseng (ginseng) have been extensively used in traditional Chinese medicine. However, herb-drug interactions between ginseng and other co-administered drugs are not fully understood concerning the effect of ginseng on drug metabolism and clearance. The current study aimed to elucidate the effect of total ginsenosides, a typical ginseng extract, on the regulation of Cyp1a2, a key enzyme to regulate drug metabolism under the normal and inflammatory conditions in mice. Female C57BL/6J mice treated with vehicle and lipopolysaccharide (LPS) were intragastrically administered ginseng extract for 7 days before hepatic P450 expression was analyzed. Primary mouse hepatocytes were also employed to further explore the effects of total ginsenosides on Cyp1a2 expression. The results showed that total ginsenosides in P.ginseng extract exhibited a concentration-dependent suppression on Cyp1a2 mRNA and protein level in both mice and primary mouse hepatocytes. Notably, the inhibitory effects of total ginsenosides on Cyp1a2 mRNA and protein expression were further enhanced following LPS treatment. Therefore, future research is warranted to investigate the role of ginsenosides in the regulation of hepatic CYP450s. Moreover, consumption of ginseng as food or supplement should be monitored for patients on combinational therapy, especially those with inflammatory diseases.

Correlation of cultivation time of Panax ginseng with metabolic profiles of nine ginsenosides and mRNA expression of genes encoding major biosynthetic enzymes

Ginseng (Panax ginseng C. A. Meyer) is one of the most important Chinese medicinal herbs. This medicinal plant produces numerous bioactive substances, including triterpene saponins of the ginsenoside-type. The endogenous levels of all types of ginsenosides largely determine the therapeutic quality of P. ginseng. Here, a field investigation was conducted to compare the changes in ginsenoside composition and mRNA expression levels of some key genes in the biosynthetic pathway of P. ginseng seedlings cultivated for different times (1, 2, 3 or 4 years). After establishing a high performance liquid chromatography method to simultaneously determine nine different ginsenosides, we measured the variations of ginsenoside contents in P. ginseng seedlings collected from our field base in Jilin province, China. The levels of total ginsenosides obviously increased with longer cultivation times and reached peak values in the 4-year seedlings. In light of the changes of ginsenosides composition, protopanaxatriol (PPT)-type ginsenosides displayed a synchronized change with total ones, while the peak point of protopanaxadiol (PPD)-type ginsenoside contents appeared in the 3-year plants. Transcript analyses of squalene synthase (SS), squalene epoxidase (SQE1) and dammarenediol synthase (DS), which are the rate-limiting enzymes in the ginsenoside biosynthetic pathway, showed that the gene expression levels of PgSS and PgSQE1 were highest in the 2-year seedlings, and could indirectly control saponins. While the PgDS gene was highly expressed in 4-year plants, and another important gene pleiotropic drug resistance (PgPDR) has a consistent trend. The further correlation analysis between gene expressions and metabolic changes showed that PgDS transcript level significantly correlated with PPT-type ginsenosides, Rg1, Re, and Rf. PgPDR expression levels positively correlated with total ginsenoside, Rg1, Re, and Rb1 (p < 0.01) and Rg3 negatively correlated with PgDS and PgPDR genes expression (p < 0.01). However, the PgSS gene expression was negatively correlated with Rb1 (p < 0.05). In conclusion, we propose that PPT-type ginsenoside and PPD or dammarenediol-II compounds have a close relationship. Particularly, we highlight recent reports on functional characterization of key genes dedicated to the production of ginsenosides in P. ginseng seedling with cultivated time.

Elicitors' influenced differential ginsenoside production and exudation into medium with concurrent Rg3/Rh2 panaxadiol induction in Panax quinquefolius cell suspensions.

Cobalt nitrate, nickel sulphate, hydrogen peroxide, sodium nitroprusside, and culture filtrates of Pseudomonas monteili, Bacillus circularans, Trichoderma atroviridae, and Trichoderma harzianum were tested to elicit ginsenoside production in a cell suspension line of Panax quinquefolius. Abiotic elicitors preferentially increased panaxadiols whereas biotic elicitors upregulated the panaxatriol synthesis. Cobalt nitrate (50μM) increased total ginsenosides content by twofold (54.3mg/L) within 5days. It also induced the Rc synthesis that was absent in the control cultures. Elicitation with P. monteili (2.5% v/v, 5days) also supported 2.4-fold enhancement in saponin yield. Elicitation by T. atroviridae or hydrogen peroxide induced the synthesis of Rg3 and Rh2 that are absent in ginseng roots. The highest ginsenosides productivity (3.2-fold of control) was noticed in cells exposed to 1.25% v/v dose of T. atroviridae for 5days. Treating cells with T. harzianum for 15days afforded maximum synthesis and leaching (8.1mg/L) of ginsenoside Rh1.

Changes in the Ginsenoside Content During Fermentation Using an Appliance for the Preparation of Red Ginseng.

The total amount of ginsenoside in fermented red ginseng (FRG) is increased by microbial fermentation. The aim of this study was to evaluate whether fermentation time and temperature affect the ginsenoside content during fermentation using an appliance for the preparation of red ginseng. The FRG and fermented red ginseng extracts (FRG-e) were prepared using an appliance for the preparation of red ginseng. The temperature was recorded and time points for sampling were scheduled at pre-fermentation (0[Formula: see text]h) and 18, 36, 48, 60 and 72[Formula: see text]h after the addition of the microbial strains. Samples of FRG and FRG-e were collected to identify changes in the ginsenoside contents at each time point during the fermentation process. The ginsenoside content was analyzed using high performance liquid chromatography (HPLC). The levels of ginsenoside Rh1, Rg3, and compound Y, which are known to have effective pharmacological properties, increased more than three-fold in the final products of FRG relative to samples prior to fermentation. Although the ginsenoside constituents of FRG-e decreased or increased and then decreased during fermentation, the total amount of ginsenoside in FRG-e was even higher than those in FRG; the total amounts of ginsenoside in FRG-e and FRG were 8282.8 and 738.0[Formula: see text]mg, respectively. This study examined the changes in composition of ginsenosides and suggests a method to manufacture high-content total ginsenosides according to the fermentation temperature and process time. Reducing the extraction time is expected to improve the decrease of ginsenosides in FRG-e as a function of the fermentation time.

Microbial conversion of major ginsenosides in ginseng total saponins by Platycodon grandiflorum endophytes

BackgroundIn this study, we screened and identified an endophyte JG09 having strong biocatalytic activity for ginsenosides from Platycodon grandiflorum, converted ginseng total saponins and ginsenoside monomers, determined the source of minor ginsenosides and the transformation pathways, and calculated the maximum production of minor ginsenosides for the conversion of ginsenoside Rb1 to assess the transformation activity of endophyte JG09. MethodsThe transformation of ginseng total saponins and ginsenoside monomers Rb1, Rb2, Rc, Rd, Rg1 into minor ginsenosides F2, C-K and Rh1 using endophyte JG09 isolated by an organizational separation method and Esculin-R2A agar assay, as well as the identification of transformed products via TLC and HPLC, were evaluated. Endophyte JG09 was identified through DNA sequencing and phylogenetic analysis. ResultsA total of 32 β-glucosidase-producing endophytes were screened out among the isolated 69 endophytes from P. grandiflorum. An endophyte bacteria JG09 identified as Luteibacter sp. effectively converted protopanaxadiol-type ginsenosides Rb1, Rb2, Rc, Rd into minor ginsenosides F2 and C-K, and converted protopanaxatriol-type ginsenoside Rg1 into minor ginsenoside Rh1. The transformation pathways of major ginsenosides by endophyte JG09 were as follows: Rb1→Rd→F2→C-K; Rb2→C-O→C-Y→C-K; Rc→C-Mc1→C-Mc→C-K; Rg1→Rh1. The maximum production rate of ginsenosides F2 and C-K reached 94.53% and 66.34%, respectively. ConclusionThis is the first report about conversion of major ginsenosides into minor ginsenosides by fermentation with P. grandiflorum endophytes. The results of the study indicate endophyte JG09 would be a potential microbial source for obtaining minor ginsenosides.

혼합용매를 이용한 진생베리 추출물 최적 공정 개발 및 추출물의 진세노사이드 분석

Abstract Ginsenosides in ginseng berry has been known as functional materials showing physiological effect to thehuman. Specially, ginseng berry contains plenty of ginsenoside Re, but the study of extraction processes were not enough performed. Accordingly, the purpose of this study was to establish the optimized extraction process for obtaining ginsenoside Re from ginseng berry. The extraction process of ginsenosides was performed in 250 mL extraction flask containing 150 solvent and 10 g of dried ginseng berry. The extracted ginsenoside Re, Rg1 and Rd and total crude ginsenosides from ginseng berry were evaluated by TLC according to the treated conditions (the ratioof alcohol to water, extraction temperature, extraction period, and extraction times). Optimized conditions for extraction was 70% to 30% of the ratio of alcohol to water, 80oC of extraction temperature, 4 h of extraction period,and 2 times of extraction frequency. The amount of total crude ginsenosides of the extract obtained from the optimized process was 88.6 mg/g based on dried ginseng berry. The composition of ginsenosides from the extracted was 5.5% of Rb1, 5.2% of Rc, 14.3% of Rd, 51.5% of Re, 8.1% of Rf, and 15.7% of Rg1. A protopanaxtriol ginsenosides of whole ginsenosides extracted was about 80%.

A Continuous Fractionation of Ginsenosides and Polysaccharides FromPanax ginsengUsing Supercritical Carbon Dioxide Technology

A continuous supercritical carbon dioxide (SC-CO2) fractionation was conducted to obtain ginsenosides and polysaccharides from Panax ginseng extract. P. ginseng was mixed with ethanol at a ratio of 1:4 (volume by weight) for 24 h to yield ethanol extract. SC-CO2 fractionation was performed using an orthogonal design to obtain optimal fractionation parameters. The conditions were: three different temperatures (40, 50 and 60C), three different pressure levels (10, 20 and 30 MPa), three flow rates of SC-CO2 (2, 4 and 6 mL/min) and three flow rates of ginseng extract liquor (1, 2 and 3 mL/min). The highest concentration of total ginsenosides was obtained in the fraction (F) whereas the highest concentration of total polysaccharides was retained in the residual. The highest selectivity of ginsenosides and polysaccharides were obtained at 60C and 30 MPa. The greatest free radicals scavenging ability and antioxidant ability were obtained in F. Practical Applications Ginsenosides and polysaccharides from Panax ginseng have been postulated to contribute a number of positive health effects. Traditional extraction techniques are associated with the disadvantages of solvent residues and its associated toxicity. In the present study, a continuous preparation method using supercritical fluid technology was described with the optimal temperature and pressure for the separation of P. ginseng ginsenosides and polysaccharide. Our findings can be served as a basis to facilitate future scale-up industrial applications.

Changes of Fatty Acids, Minerals and Ginsenosides on Ginseng Seeds during Stratifying Treatment

Background : This study was carried out to investigate the changes to fatty acid, mineral, and ginsenosides contents in ginseng seed when they were stratified for different length of time and to determine whether variety had any effects on the changes. The aim was to improve the ginseng seed stratification process. Methods and Results : The ginseng varieties used were Geumpoong, Chunpoong, Yunpoong, and K-1. Stratifying periods treated on ginseng seed were 0, 20, 40, 60, 80, and 100 days. The main fatty acids of ginseng seed were oleic acid (C18 : 1, n9c) with a content of 78.40 - 79.20% followed by linoleic acid (C18 : 2, n6c). The main mineral in the seeds was potassium (K), at 1208.2 - 1337.6 ㎎/100 g. The main ginsenosides in ginseng seed were ginsenoside Re and Rb1. Increasing the length of the stratification periods led to increases in oleic acid content (60 - 80 days), however after this the content declined. In contrast, linoleic acid content fell as the stratification period increased. K, P, Mg, Ca and Na content rose as the stratification period increased. The ginsenoside Re content of Chunpoong and K-1 cultivar seeds also rose as the stratification period increased which meant that total ginsenoside content increased. However, ginsenoside Re content rose in Geumpoong and Yunpoong seeds, but total ginsenoside content decreased as the stratification period increased. Conclusions : Some beneficial compound in ginseng seed rose as the stratification period increased. Therefore, ginseng seed stratification could improve the food value of ginseng.

Total ginsenosides suppress monocrotaline-induced pulmonary hypertension in rats: involvement of nitric oxide and mitogen-activated protein kinase pathways

BackgroundGinsenosides have been shown to exert beneficial pharmacological effects on the central nervous, cardiovascular, and endocrine systems. We sought to determine whether total ginsenosides (TG) inhibit monocrotaline (MCT)-induced pulmonary hypertension and to elucidate the underlying mechanism. MethodsMCT-intoxicated rats were treated with gradient doses of TG, with or without NG-nitro-l-arginine methyl ester. The levels of molecules involving the regulation of nitric oxide and mitogen-activated protein kinase pathways were determined. ResultsTG ameliorated MCT-induced pulmonary hypertension in a dose-dependent manner, as assessed by the right ventricular systolic pressure, the right ventricular hypertrophy index, and pulmonary arterial remodeling. Furthermore, TG increased the levels of pulmonary nitric oxide, endothelial nitric oxide synthase, and cyclic guanosine monophosphate. Lastly, TG increased mitogen-activated protein kinase phosphatase-1 expression and promoted the dephosphorylation of extracellular signal-regulated protein kinases 1/2, p38 mitogen-activated protein kinase, and c-Jun NH2-terminal kinase 1/2. ConclusionTG attenuates MCT-induced pulmonary hypertension, which may involve in part the regulation of nitric oxide and mitogen-activated protein kinase pathways.

Research of chemotaxis response of Botrytis cinerea and Alternaria panax on total ginsenosides

In this paper, three kinds of chemotactic parameters (concentration, temperature and pH) were determined by plate assay and spore germination method to research the chemotactic response of Botrytis cinerea and Alternaria panax, and their spores on total ginsenosides. The results showed that Botrytis cinerea had strong chemotactic response at the mid-concentration of total ginsenosides (cultivation temperature was 20 degrees C and pH value was 6), and the data of chemotactic migration index (CMI) was 1.293 0, chemotactic growth rate (CGR) was 0.476 0, spore germination rate (SGR) was 53%, and dry weight of mycelial (DWM) was 0.452 6 g x L(-1); however, Alternaria panax had strong chemotactic response at the low-concentration of total ginsenosides (cultivation temperature was 25 degrees C and pH value was 6), and the data of chemotactic migration index (CMI) was 1.235 4, chemotactic growth rate (CGR) was 0.537 0, spore germination rate (SGR) was 67%, and dry weight of mycelial (DWM) was 0.494 8 g x L(-1). The results indicated that the low and middle concentration (2, 20 mg x L(-1)) of total ginsenosides had significant promoting effect on chemotactic response of these two pathogens, and the spore germination, mycelial growth rate, dry weight of mycelial of them were also significantly improved by this chemotactic response, whereas it decreased as the increase of total ginsenosides concentration.

A green protocol for efficient discovery of novel natural compounds: Characterization of new ginsenosides from the stems and leaves of Panax ginseng as a case study

Exploration of new natural compounds is of vital significance for drug discovery and development. The conventional approaches by systematic phytochemical isolation are low-efficiency and consume masses of organic solvent. This study presents an integrated strategy that combines offline comprehensive two-dimensional liquid chromatography, hybrid linear ion-trap/Orbitrap mass spectrometry, and NMR analysis (2D LC/LTQ-Orbitrap-MS/NMR), aimed to establish a green protocol for the efficient discovery of new natural molecules. A comprehensive chemical analysis of the total ginsenosides of stems and leaves of Panax ginseng (SLP), a cardiovascular disease medicine, was performed following this strategy. An offline 2D LC system was constructed with an orthogonality of 0.79 and a practical peak capacity of 11,000. The much greener UHPLC separation and LTQ-Orbitrap-MS detection by data-dependent high-energy C-trap dissociation (HCD)/dynamic exclusion were employed for separation and characterization of ginsenosides from thirteen fractionated SLP samples. Consequently, a total of 646 ginsenosides were characterized, and 427 have not been isolated from the genus of Panax L. The ginsenosides identified from SLP exhibited distinct sapogenin diversity and molecular isomerism. NMR analysis was finally employed to verify and offer complementary structural information to MS-oriented characterization. The established 2D LC/LTQ-Orbitrap-MS/NMR approach outperforms the conventional approaches in respect of significantly improved efficiency, much less use of drug materials and organic solvent. The integrated strategy enables a deep investigation on the therapeutic basis of an herbal medicine, and facilitates new compounds discovery in an efficient and environmentally friendly manner as well.

A UHPLC–TOF/MS method based metabonomic study of total ginsenosides effects on Alzheimer disease mouse model

A metabonomic method was established to find potential biomarkers and study the metabolism disturbance in Alzheimer disease animal model. Total ginsenosides, as potential agent in neuroprotection and anti-inflammation, was also studied to learn the regulation mechanism to plasma metabolites in model animals.In experiment, amyloid beta 1–42 was occupied to form Alzheimer disease animal model. After drug administration, animals were evaluated by Morris water maze behavior test and sacrificed. Plasma samples were then analyzed using UHPLC–TOF/MS method to determine the endogenous metabolites.Behavior test results revealed that the spatial learning and memory abilities were deficit in model mice, and total ginsenosides could improve cognition abilities in dose-dependent manners. Principal component analysis showed that model and sham were divided into two groups, which means the metabolic network of mice was disturbed after modeling. Accordingly, 19 biomarkers were found and identified. In model group, the levels of proline, valine, tryptophan, LPC (14:0), LPC (15:0), LPC (15:1), LPC (17:0), LPC (18:2), LPC (18:3) and LPC (20:4) were up-regulated, while the levels of acetylcarnitine, palmitoylcarnitine, vaccenylcarnitine, phytosphingosine, N-eicosanoylethanolamine, hexadecenoic acid, docosahexaenoic acid, docosapentaenoic acid and octadecadienoic acid were down-regulated. The levels of these metabolites were recovered in different degrees after total ginsenosides administration. Combining with behavior study results, total ginsenosides could ameliorate both cognition symptoms and metabolic changes in model animals. This metabonomic approach provided a feasible way to understand the endogenous alterations of AD and to study the pharmacodynamic activity of novel agents.