Ginsenoside F2 Research Articles

Computer-Guided Engineered Endo- and Exocleaving Glycosidase for Significantly Improving Production of Ginsenoside F1.

Ginsenoside F1, a particularly rare and valuable compound known for its health benefits, requires precise deglycosylation due to the extensive glycosylation of ginsenosides in Panax notoginseng. Here, we identified that the β-d-glucosidase BglSK exhibits both endo- and exocleaving glycosidase activities with multi-6-O-glycosides, thereby facilitating the specific production of Ginsenoside F1. The variant BglSKT137A/L508A, obtained through protein engineering, displayed kcat/KM values for the reactions of ginsenoside Rg1 and notoginsenoside R1 that were increased by 13.88-fold and 108.56-fold, respectively, compared with the BglSKWT. The reduced steric hindrance and the overall increase in loop stability show a higher tendency to adopt a closed conformation and facilitate the prereaction state, which may explain the enhanced catalytic efficiency of the engineered enzyme. These beneficial mutants will deepen our understanding of mechanisms for improving glycosidase activity and provide tools for producing high-value P. notoginseng products.

Ginseng fruit rare saponins (GFRS) improved inflammatory response: In vitro and in vivo assessment

Inflammation is the body's protective immune response to tissue damage. Ginseng has a long history of medicinal use, and its active ingredient ginsenosides have anti-inflammatory effects. Ginseng fruit rare saponins (GFRS) is a transformation product of ginseng saponins and rich in a variety of rare saponins. We used HPLC-DAD method to study GFRS rare saponins with ginsenoside F4, R-Rg3, SRg3, Rk1, Rg6, Rg5, Rk3 and Rh4. However, there is no study on the use of GFRS to reduce skin inflammation. This study enriched the action pathway of GFRS through network pharmacology and revealed the anti-inflammatory effect of GFRS for the first time. In vitro experiments showed that GFRS could significantly reduce the release of NO in lipopolysaccharide (LPS) -induced RAW264.7 cells and HaCaT cells, and reduce the secretion and expression of inflammation-related factors Interleukin-6 (IL-6), Tumor necrosis factor-α (TNF-α) and Interleukin-17 A (IL-17 A), thereby reducing cell inflammatory damage. In the imiquimod (IMQ) -induced mouse inflammatory model, the therapeutic effect of GFRS on the pathogenesis of psoriasis-like dermatitis was studied. In vivo experiments showed that the skin erythema, scales, thickness and inflammatory infiltration of GFRS-treated mice were reduced, and the psoriasis area severity index score was significantly lower than that of IMQ group. GFRS restored IMQ-induced spleen size and reduced the secretion and expression of TNF-α, IL-6, Interferon-γ (IFN-γ) and IL-17 A in serum. In summary, our results demonstrate that GFRS alleviates IMQ-induced dermatitis symptoms, effectively reduces the secretion of inflammatory factors, and inhibits IL-17 A expression.

Anti-Warburg Mechanism of Ginsenoside F2 in Human Cervical Cancer Cells via Activation of miR193a-5p and Inhibition of β-Catenin/c-Myc/Hexokinase 2 Signaling Axis.

Though Ginsenoside F2 (GF2), a protopanaxadiol saponin from Panax ginseng, is known to have an anticancer effect, its underlying mechanism still remains unclear. In our model, the anti-glycolytic mechanism of GF2 was investigated in human cervical cancer cells in association with miR193a-5p and the β-catenin/c-Myc/Hexokinase 2 (HK2) signaling axis. Here, GF2 exerted significant cytotoxicity and antiproliferation activity, increased sub-G1, and attenuated the expression of pro-Poly (ADPribose) polymerase (pro-PARP) and pro-cysteine aspartyl-specific protease (procaspase3) in HeLa and SiHa cells. Consistently, GF2 attenuated the expression of Wnt, β-catenin, and c-Myc and their downstream target genes such as HK2, pyruvate kinase isozymes M2 (PKM2), and lactate dehydrogenase A (LDHA), along with a decreased production of glucose and lactate in HeLa and SiHa cells. Moreover, GF2 suppressed β-catenin and c-Myc stability in the presence and absence of cycloheximide in HeLa cells, respectively. Additionally, the depletion of β-catenin reduced the expression of c-Myc and HK2 in HeLa cells, while pyruvate treatment reversed the ability of GF2 to inhibit β-catenin, c-Myc, and PKM2 in GF2-treated HeLa cells. Notably, GF2 upregulated the expression of microRNA139a-5p (miR139a-5p) in HeLa cells. Consistently, the miR139a-5p mimic enhanced the suppression of β-catenin, c-Myc, and HK2, while the miR193a-5p inhibitor reversed the ability of GF2 to attenuate the expression of β-catenin, c-Myc, and HK2 in HeLa cells. Overall, these findings suggest that GF2 induces apoptosis via the activation of miR193a-5p and the inhibition of β-catenin/c-Myc/HK signaling in cervical cancer cells.

새싹인삼 함유 콤부차의 영양성분 및 생리활성

In this study, the physicochemical properties, nutritional components, and antioxidant activities of kombucha containing ginseng sprouts (control kombucha, CT; strawberry kombucha, ST ; strawberry kombucha with 2% ginseng sprout, ST+GS) were analyzed for comparison of quality characteristics. The total content of free amino acids in ST+GS (273.38 mg/100 mL) was 3.2-14.5 times higher than in CT (18.9 mg/100 mL) and ST (84.9 mg/100 mL). The total mineral content in ST+GS (63.99 mg/100 mL) was 3.3-4.1 times higher than those of CT and ST (15.45 and 19.28 mg/100 mL). The contents of soluble phenolic and soluble flavonoid were 1.2 mg GAE/mL and 0.14 mg RE/mL in ST+GS. Several ginsenosides were detected only in ST+GS; ginsenoside Rg2 (2.4 mg/100 mL), Rh1 (4.5 mg/100 mL), F2 (9.0 mg/100 mL), Rg3 (4.6 mg/100 mL), and compound K (7.8 mg/100 mL) were detected. The content of phenolic acids was 1.2-1.5 times higher in ST+GS than in CT and ST. The amount of flavonol of ST+GS was not significantly different from CT but was 1.4 times higher than in ST. In terms of antioxidant activities, the values of ST+GS were significantly higher in comparison to other kombucha samples. These results confirmed that incorporating ginseng sprouts amplifies the advantages of kombucha.

새싹인삼 함유 콤부차의 영양성분 및 생리활성

In this study, the physicochemical properties, nutritional components, and antioxidant activities of kombucha containing ginseng sprouts (control kombucha, CT; strawberry kombucha, ST ; strawberry kombucha with 2% ginseng sprout, ST+GS) were analyzed for comparison of quality characteristics. The total content of free amino acids in ST+GS (273.38 mg/100 mL) was 3.2-14.5 times higher than in CT (18.9 mg/100 mL) and ST (84.9 mg/100 mL). The total mineral content in ST+GS (63.99 mg/100 mL) was 3.3-4.1 times higher than those of CT and ST (15.45 and 19.28 mg/100 mL). The contents of soluble phenolic and soluble flavonoid were 1.2 mg GAE/mL and 0.14 mg RE/mL in ST+GS. Several ginsenosides were detected only in ST+GS; ginsenoside Rg2 (2.4 mg/100 mL), Rh1 (4.5 mg/100 mL), F2 (9.0 mg/100 mL), Rg3 (4.6 mg/100 mL), and compound K (7.8 mg/100 mL) were detected. The content of phenolic acids was 1.2-1.5 times higher in ST+GS than in CT and ST. The amount of flavonol of ST+GS was not significantly different from CT but was 1.4 times higher than in ST. In terms of antioxidant activities, the values of ST+GS were significantly higher in comparison to other kombucha samples. These results confirmed that incorporating ginseng sprouts amplifies the advantages of kombucha.

Revealing the Mechanisms of Qilongtian Capsules in the Treatment of Chronic Obstructive Pulmonary Disease Based on Integrated Network Pharmacology, Molecular Docking, and In Vivo Experiments.

The Qilongtian capsule (QLT) is a Chinese patent medicine that has been approved for the treatment of chronic obstructive pulmonary disease (COPD). However, the precise pharmacodynamic material basis and molecular mechanism have not been well illustrated. In this study, we identified the effect of QLT on COPD through a cigarette smoke extract (CSE)/lipopolysaccharide (LPS) induced COPD mice model. The absorption of blood components in QLT were identified using ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Network pharmacology was used to predict the potential targets and therapeutic mechanisms of QLT, which were further validated using in vivo experiments and molecular docking. Pharmacodynamic studies revealed that QLT could ameliorate pulmonary function and pulmonary pathology, reduce collagen fiber accumulation, and attenuate inflammatory responses in mice with CSE/LPS induced COPD. A total of 21 components of QLT absorbed in the blood were detected. Network pharmacology analysis indicated that TNF, IL-6, EGFR, and AKT1 may be the core targets, mainly involving the MAPK signaling pathway. Besides, Sachaloside II, Ginsenoside Rh1, Ginsenoside F1, Rosiridin, and Ginsenoside Rf were the key compounds. Molecular docking results showed that the key components could spontaneously bind to EGFR and MAPK to form a relatively stable conformation. In vivo experiments revealed that QLT could suppress the activation of the EGFR/MAPK signaling pathway, thereby improving lung injury in mice with COPD. Overall, these findings provide evidence for the treatment of COPD with QLT.

Investigating alterations of nutritional constituents, antioxidant abilities, and physicochemicals in aging processes of ginseng sprouts

This study was the first to document the fluctuations of nutritional constituents, antioxidant capacities, and physicochemical characteristics during the aging processes of red ginseng sprouts (RGS) and black ginseng sprouts (BGS) from dried ginseng sprouts (DGS). Total ginsenoside levels differed with 54.72 (DGS) → 57.15 (RGS) and 6.98 (BGS) mg/g, specifically, ginsenoside F2 and Rd2 in RGS remarkably increased with 1.97 → 5.88 and 2.41 → 5.49 mg/g, respectively. Phenolics increased dramatically as 297.02 → 1770.01 (6.0-fold); 1834.94 (6.2-fold) μg/g in DGS → RGS; BGS with abundance contents of benzoic acid (>15.3-fold), chlorogenic acid (>9.5-fold), and catechin (>4.2-fold), whereas amino acids markedly decreased (3686.81 → 1505.00; 364.64 mg/100 g), with arginine showing a significant decrease. Moreover, beneficial factors (total phenolic content: TPC; total flavonoid content: TFC; maillard reaction products: MRP) displayed increase tendencies (approximately 2.0-fold) with BGS > RGS > DGS, and antioxidant patterns significantly increased with potential capacities as follows: ABTS (48.3: DGS → 65.8: RGS; 70.2 %: BGS) > DPPH (18.5 → 44.6; 59.2 %) > hydroxyl (23.2 → 35.4; 39.9 %) > FRAP (0.6 → 1.8; 1.8 %) at 500 μg/mL. In particular, DNA protection exhibited excellent rates of 100 %, in the order of BGS (25 μg/mL) > RGS (50 μg/mL) > DGS (500 μg/mL). These findings suggest that processed ginseng sprouts can be excellent agents for natural antioxidants.

Heterologous synthesis of ginsenoside F1 and its precursors in Nicotiana benthamiana

Ginsenoside F1 has high medicinal values, which is a kind of rare triterpene saponin isolated from Panax plants. The extremely low content of ginsenoside F1 in herbs has limited its research and application in medical field. In this work, we constructed a pathway in tobacco for the biosynthesis of ginsenoside F1 by metabolic engineering. Four enzyme genes (PnDDS, CYP716A47, CYP716S1 and UGT71A56) isolated from Panax notoginseng were introduced into tobacco. Thus, a biosynthetic pathway for ginsenoside F1 synthesis was artificially constructed in tobacco cells; moreover, the four exogenous genes could be expressed in the roots, stems and leaves of transgenic plants. Consequently, ginsenoside F1 and its precursors were successfully synthesized in the transgenic tobacco, compared with Panax plants, the content of ginsenoside F1 in transgenic tobacco was doubled. In addition, accumulation of ginsenoside F1 and its precursors in transgenic tobacco shows organ specificity. Based on these results, a new approach was established to produce rare ginsenoside F1; meanwhile, such strategy could also be employed in plant hosts for the heterologous synthesis of other important or rare natural products.

Revealing the pharmacological mechanisms of nao-an dropping pill in preventing and treating ischemic stroke via the PI3K/Akt/eNOS and Nrf2/HO-1 pathways.

Nao-an Dropping Pill (NADP) is a Chinese patent medicine which commonly used in clinic for ischemic stroke (IS). However, the material basis and mechanism of its prevention or treatment of IS are unclear, then we carried out this study. 52 incoming blood components were resolved by UHPLC-MS/MS from rat serum, including 45 prototype components. The potential active prototype components hydroxysafflor yellow A, ginsenoside F1, quercetin, ferulic acid and caffeic acid screened by network pharmacology showed strongly binding ability with PIK3CA, AKT1, NOS3, NFE2L2 and HMOX1 by molecular docking. In vitro oxygen-glucose deprivation/reperfusion (OGD/R) experimental results showed that NADP protected HA1800 cells from OGD/R-induced apoptosis by affecting the release of LDH, production of NO, and content of SOD and MDA. Meanwhile, NADP could improve behavioral of middle cerebral artery occlusion/reperfusion (MCAO/R) rats, reduce ischemic area of cerebral cortex, decrease brain water and glutamate (Glu) content, and improve oxidative stress response. Immunohistochemical results showed that NADP significantly regulated the expression of PI3K, Akt, p-Akt, eNOS, p-eNOS, Nrf2 and HO-1 in cerebral ischemic tissues. The results suggested that NADP protects brain tissues and ameliorates oxidative stress damage to brain tissues from IS by regulating PI3K/Akt/eNOS and Nrf2/HO-1 signaling pathways.

Ginseng fruit rare saponins (GFRS), a promising anti-wrinkle agent: Evidence of its antioxidant effect and its capacity to prevent matrix metalloproteinase (MMPs) expression in vitro and in vivo

Rare saponins show numerous significant biological activities, however, there has been no research on using ginseng fruit rare saponins (GFRS) to minimize skin wrinkles. This paper studied the anti-wrinkle effect, related mechanism, material basis of efficacy, and structure-activity relationship of GFRS in vivo and in vitro. The findings demonstrate that GFRS has potent collagenase and elastase inhibition capabilities in addition to superior ultraviolet absorption properties. Secondly, GFRS can strengthen the endogenous antioxidant defense system, thus lessening the UVB-induced ultraviolet damage that L929 cells and zebrafish experience. Matrix metalloproteinases (MMPs) studies have shown that GFRS could significantly inhibit the UVB-induced MMP1 and MMP9 proteins production. Finally, the main ginsenoside component in GFRS was identified by LC-MS, and 16 kinds of ginsenoside were analyzed, including rare ginsenoside such as Rg6, F4, and R/S-Rg3, which may be caused by the transformation of prototype saponins Re and Rd caused by citric acid heat treatment (CAHT). After doing a quantitative HPLC analysis on twelve of them, it was discovered that CAHT could enrich the content of rare saponins in GFS. By evaluating these ginsenoside’s anti-wrinkle properties. It is found that the main material basis for GFRS’s anti-wrinkle action may be ginsenoside Rg6 or F4. In summary, GFRS exhibits promising properties as an anti-wrinkle treatment because of its robust anti-oxidation, anti-collagenase, anti-elastase activity, favorable ultraviolet absorption effect, and capacity to regulate MMPs in vivo and in vitro models.

Targeted preparation of six rare ginsenosides using two β-glucosidases from Bifidobacterium adolescentis

In this study, six rare protopanaxadiol (PPD)-type ginsenosides, namely, F2, CK, C-Mc1, C-Mc, CO, and CY, were prepared using enzymatic hydrolysis. Firstly, major (PPD)-type ginsenosides, including Rb1, Rb2, Rc, and Rd, were biotransformed by employing single and combined β-glucosidases BaBgl1A and BaBgl3A. Thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analyses revealed that ginsenosides F2, CO, and C-Mc1 could be produced by BaBgl1A, while rare ginsenosides CK, CY, and C-Mc could be produced by the combined use of BaBgl1A and BaBgl3A. Furthermore, optimal preparation conditions for these rare ginsenosides were elucidated, with yields exceeding 76.34%. Additionally, large quantities of rare ginsenosides were prepared to determine their structures. Characteristic absorption peaks in 13C NMR spectra and ion peaks in MALDI-TOF-MS suggested that the six high-purity rare ginsenosides (i.e., F2, CO and C-Mc1, CK, CY, and C-Mc) could be produced at high-efficiency. The findings of this study offer insights into novel enzymolysis methods for the targeted preparation of rare PPD-type ginsenosides, thus widening the range of raw materials that could be directed to in-depth biological activity research.

Protective effects of ginsenoside F2 on isoproterenol-induced myocardial infarction by activating the Nrf2/HO-1 and PI3K/Akt signaling pathways

BackgroundGinsenoside F2 (GF2) serves as the principal intestinal metabolite resulting from the oral intake of Panax ginseng and Panax quinquefolius, exhibiting antioxidative, hypolipidemic, antitumor, and anti-inflammatory properties. Nevertheless, its effect on myocardial infarction (MI) is still unknown. PurposeThe purpose of this study is to investigate the protective effect and the underlying mechanisms of GF2 against isoproterenol (ISO)-induced MI. MethodsISO-induced H9c2 cardiomyocytes and MI rat models were utilized as in vitro and in vivo models to evaluate the impact of anti-MI of GF2. The underlying mechanisms were investigated using a variety of methodologies, including electrocardiography, Western blot analysis, histopathological examination, immunofluorescence, immunohistochemistry, and ELISA techniques. ResultsIn vivo experiments, our results indicated that GF2 significantly ameliorated ISO-induced electrocardiographic (ECG) abnormalities, myocardial fiber necrosis, rupture, fibrosis of myocardial tissues, and suppressed cardiac enzyme activities. Meanwhile, GF2 notably raised the activity of antioxidant enzymes like CAT, GSH, and SOD. Furthermore, it downregulated Keap1 expression level while upregulating NQO1, Nrf2, and HO-1 expression levels. Additionally, GF2 suppressed the expression of the cleaved caspase-3 and pro-apoptotic protein Bax while promoting the expression of anti-apoptotic proteins Bcl-2, p-PI3K, and p-Akt. TUNEL fluorescence results also demonstrated that GF2 effectively inhibited cardiomyocyte apoptosis. Furthermore, consistent with the results of animal experiments, GF2 considerably attenuated ROS generation, changed apoptosis and mitochondrial function, and reduced oxidative stress in ISO-induced H9c2 cardiomyocytes through activating Nrf2/HO-1 and PI3K/Akt signaling pathways. ConclusionTaken together, GF2 ameliorated MI by preventing cardiocyte apoptosis, oxidative stress, and mitochondrial dysfunction via modulating the Nrf2/HO-1 and PI3K/Akt signaling pathways, showing potential as a treatment strategy for treating MI.

Enzymatic pretreatment of Panax notoginseng leaves alleviates obesity and modulates the gut microbiota in high-fat diet-fed mice

The root of Panax notoginseng (Burk.) F.H. is one of the most famous natural medicines and functional foods for numerous diseases. The saponin constituents present in Panax notoginseng leaves (PNGL) demonstrate an elevated level of glycosylation in comparison to those in the roots, resulting in a significantly diminished bioavailability. In this study, we systematically screened nine recombinant thermostable glycosidases and identified a synergistic combination, namely Tpexy3 and Tpebgl3, to catalyze PNGL. At 70 °C, 2000 U of both glycosidases were added before and after 1 g of PNGL, yielding 650 mg of new fractions, of which ginsenosides Rg3, F2, CK, and PPD were identified as the major products. The enzymatic PNGL demonstrated a lipid droplet-lowering effect on the 3 T3-L1 lipid deposition model. Furthermore, the enzymatic PNGL exhibited significant inhibitory effects on weight gain, hyperlipidemia, hyperglycemia, and inflammation in high-fat diet (HFD) mice. Importantly, the enzymatic PNGL restored the HFD-induced increase in the Firmicutes/Bacteroidetes (F/B) ratio and promoted the growth of beneficial gut microbes with anti-inflammatory properties, improved lipid metabolism, and short-chain fatty acid production. These results strongly indicate that enzyme-catalyzed pretreatment of PNGL can effectively enhance its bioavailability and anti-obesity effects. Biological enzyme catalysis not only provides technical support for the development of specific functional products but also promotes the efficient and high-value utilization of natural product resources.

Biotransformation approach to produce rare ginsenosides F1, compound Mc1, and Rd2 from major ginsenosides.

The stems and leaves of Panax notoginseng contain high saponins, but they are often discarded as agricultural waste. In this study, the predominant ginsenosides Rg1, Rc, and Rb2, presented in the stems and leaves of ginseng plants, were biotransformed into value-added rare ginsenosides F1, compound Mc1 (C-Mc1), and Rd2, respectively. A fungal strain YMS6 (Penicillium sp.) was screened from the soil as a biocatalyst with high selectivity for the deglycosylation of major ginsenosides. Under the optimal fermentation conditions, the yields of F1, C-Mc1, and Rd2 were 97.95, 68.64, and 79.58%, respectively. This study provides a new microbial resource for the selective conversion of protopanaxadiol-type and protopanaxatriol-type major saponins into rare ginsenosides via the whole-cell biotransformation and offers a solution for the better utilization of P. notoginseng waste.

Enhanced brain distribution of Ginsenoside F1 via intranasal administration in combination with absorption enhancers

Ginsenoside F1 (GF1) is a potential drug candidate for the treatment of Alzheimer's disease. Nevertheless, its low oral bioavailability and poor solubility limit clinical application. By utilizing either a direct or indirect approach, intranasal administration is a non-invasive drug delivery method that can deliver drugs to the brain rapidly. But large molecule drug delivered to the brain through intranasal administration may be insufficient to reach required concentration for therapeutic effect. In this study, using GF1 as a model drug, the feasibility of intranasal administration in combination with absorption enhancers to increase brain distribution of GF1 was explored. First of all, the appropriate absorption enhancers were screened by in situ nasal perfusion study. GF1-HP-β-CD inclusion complex was prepared and characterized. Thereafter, in vivo absorption of GF1 after intranasal or intravenous administration of its inclusion complex with/without absorption enhancers was investigated, and safety of the formulations was evaluated. The results showed that 2% Solutol HS 15 was a superior absorption enhancer. HP-β-CD inclusion complex improved GF1 solubility by 150 fold. Following intranasal delivery, the absolute bioavailability of inclusion complex was 46%, with drug brain targeting index (DTI) 247% and nose-to-brain direct transport percentage (DTP) 58%. Upon further addition of 2% Solutol HS 15, the absolute bioavailability was increased to 75%, with DTI 315% and DTP 66%. Both nasal cilia movement and biochemical substances (total protein and lactate dehydrogenase) leaching studies demonstrated 2% Solutol HS 15 was safe to the nasal mucosa. In conclusion, intranasal administration combining with safe absorption enhancers is an effective strategy to enhance drug distribution in the brain, showing promise for treating disorders related to the central nervous system.

Increasing the Amounts of Bioactive Components in American Ginseng (Panax quinquefolium L.) Leaves Using Far-Infrared Irradiation.

Both the roots and leaves of American ginseng contain ginsenosides and polyphenols. The impact of thermal processing on enhancing the biological activities of the root by altering its component composition has been widely reported. However, the effects of far-infrared irradiation (FIR), an efficient heat treatment method, on the bioactive components of the leaves remain to be elucidated. In the present study, we investigated the effects of FIR heat treatment between 160 and 200 °C on the deglycosylation and dehydration rates of the bioactive components in American ginseng leaves. As the temperature was increased, the amounts of common ginsenosides decreased while those of rare ginsenosides increased. After FIR heat treatment of American ginseng leaves at an optimal 190 °C, the highest total polyphenolic content and kaempferol content were detected, the antioxidant activity was significantly enhanced, and the amounts of the rare ginsenosides F4, Rg6, Rh4, Rk3, Rk1, Rg3, and Rg5 were 41, 5, 37, 64, 222, 17, and 266 times higher than those in untreated leaves, respectively. Moreover, the radical scavenging rates for 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) and the reducing power of the treated leaf extracts were 2.17, 1.86, and 1.77 times higher, respectively. Hence, FIR heat treatment at 190 °C is an efficient method for producing beneficial bioactive components from American ginseng leaves.

Ginsenoside F1 inhibits cholesterol overload in oxidative-damaged cells through SREBP2/HMGCR pathway

To investigate the inhibitory mechanisms of ginsenoside F1 on hydrogen peroxide induced cholesterol metabolism disorder and oxidative stress in HepG2 cells. 1, 1-diphenyl-2-picrylhydrazyl(DPPH) and oxygen radical absorbance capacity(ORAC) tests were used to detect the scavenging effect of ginsenoside F1 on nitrogen and oxygen free radicals. HepG2 cells were treated with 400 μmol/L hydrogen peroxide and pretreated with 10, 20 and 40 μmol/L ginsenoside F1. Mitochondrial membrane potential(MMP) and total cholesterol levels were detected by JC-1 method and cholesterol kit, respectively. The protein expression levels of sterol-regulatory element binding proteins(SREBP2)and 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGCR) in cholesterol synthesis pathway were detected by Western blot. The DPPH clearance rate of ginsenoside F1 was much lower than that of 6-hydroxy-2, 5, 7, 8-tetramethylchroman-2-carboxylic acid(Trolox), but the ORAC capability of ginsenoside F1 was stronger, which was comparable to Trolox. The MMP and protein expression of SREBP2 were significantly decreased in injured group(P<0.05). The cholesterol and protein expression of HMGCR were significantly increased(P<0.05). Whereas, compared with the injured group, the MMP and protein expression of SREBP2 were significantly increased after 10, 20 and 40 μmol/L ginsenoside F1 pretreatment of injured cells(P<0.05). The cholesterol level and protein expression of HMGCR were significantly lower than injured group with concentration-dependent decreases(P<0.05). Ginsenoside F1 can protect against hydrogen peroxide induced oxidative stress in HepG2 cells by inhibiting oxygen free radicals and protecting mitochondria. And its mechanism may be related to the intervention of SREBP2/HMGCR pathway in regulating cellular cholesterol anabolism.

Impact of Sodium Silicate Supplemented, IR-Treated Panax Ginseng on Extraction Optimization for Enhanced Anti-Tyrosinase and Antioxidant Activity: A Response Surface Methodology (RSM) Approach.

Ginseng has long been widely used for its therapeutic potential. In our current study, we investigated the impact of abiotic stress induced by infrared (IR) radiations and sodium silicate on the upregulation of antioxidant and anti-tyrosinase levels, as well as the total phenolic and total flavonoid contents of the Korean ginseng (Panax ginseng C.A. Meyer) variety Yeonpoong. The RSM-based design was used to optimize ultrasonic-assisted extraction time (1-3 h) and temperature (40-60 °C) for better anti-tyrosinase activity and improved antioxidant potential. The optimal extraction results were obtained with a one-hour extraction time, at a temperature of 40 °C, and with a 1.0 mM sodium silicate treatment. We recorded maximum anti-tyrosinase (53.69%) and antioxidant (40.39%) activities when RSM conditions were kept at 875.2 mg GAE/100 g TPC, and 3219.58 mg catechin/100 g. When 1.0 mM sodium silicate was added to the media and extracted at 40 °C for 1 h, the highest total ginsenoside content (368.09 mg/g) was recorded, with variations in individual ginsenosides. Ginsenosides Rb1, Rd, and F2 were significantly affected by extraction temperature, while Rb2 and Rc were influenced by the sodium silicate concentration. Moreover, ginsenoside F2 increased with the sodium silicate treatment, while the Rg3-S content decreased. Interestingly, higher temperatures favored greater ginsenoside diversity while sodium silicate impacted PPD-type ginsenosides. It was observed that the actual experimental values closely matched the predicted values, and this agreement was statistically significant at a 95% confidence level. Our findings suggest that the application of IR irradiation in hydroponic systems can help to improve the quality of ginseng sprouts when supplemented with sodium silicate in hydroponic media. Optimized extraction conditions using ultrasonication can be helpful in improving antioxidant and anti-tyrosinase activity.

Ginsenoside F2-Mediated Intestinal Microbiota and Its Metabolite Propionic Acid Positively Impact the Gut-Skin Axis in Atopic Dermatitis Mice.

Atopic dermatitis (AD) is a complex inflammatory skin disease induced by multiple factors. AD can also cause intestinal inflammation and disorders of the gut microbiota. Ginseng is a kind of edible and medicinal plant; its main active components are ginsenosides. Ginsenosides have a variety of anti-inflammatory effects and regulate the gut microbiota; however, their role in AD and the underlying mechanisms are unclear. In this study, we found that intragastric administration of ginsenoside F2 improved AD-like skin symptoms and reduced inflammatory cell infiltration, serum immunoglobulin E levels, and mRNA expression of inflammatory cytokines in AD mice. 16s rRNA sequencing analysis showed that ginsenoside F2 altered the intestinal microbiota structure and enriched the short-chain fatty acid-producing microbiota in AD mice. Metabolomic analysis revealed that ginsenoside F2 significantly increased the propionic acid (Pa) content of feces and serum in AD mice, which was positively correlated with significant enrichment of Parabacteroides goldsteinii and Lactobacillus plantarum in the intestines. Pa inhibits inflammatory responses in the gut and skin of AD mice through the G-protein-coupled receptor43/NF-κB pathway, thereby improving skin AD symptoms. These results revealed, for the first time, the mechanism by which ginsenoside F2 improves AD through the Pa (a metabolite of intestinal microbiota)-gut-skin axis.

Screening and verification of hemostatic effective components group of Panax Notoginseng based on spectrum-effect relationships

Ethnopharmacological relevancePanax Notoginseng (PN) can disperse blood stasis, hemostasis, and detumescence analgesic, which can be used for hemoptysis, hematemesis and another traumatic bleeding, and it is known as “A miracle hemostatic medicine”. Studies show that the chemical composition of PN is relatively comprehensive, however, its hemostatic active ingredients have not been fully clarified. Aim of studyThis study aimed to clarify the hemostatic effective components group (HECG) of PN, provide a foundation for the assessment of PN's quality and its comprehensive development, and for further studies on the pharmacodynamic material basis of other Traditional Chinese Medicines (TCMs). Materials and methodsUPLC-MS was used to establish the fingerprint and identify the common peaks in 44 batches of PN extracts (PNE). In addition, the plasma recalcification time and in vitro coagulation time were measured. For spectrum-effect analysis, bivariate correlation analysis (BCA) and partial least squares regression analysis (PLSR) were used to screen the hemostasis candidate active monomers of PN. The monomers were prepared by combining several preparative chromatography techniques. The efficacy was verified by plasma recalcification time, in vitro coagulation time, and a rat model of gastric hemorrhage. ResultsA total of 30 common peaks and hemostatic efficacy indexes of 44 batches of PNE were obtained. A total of 18 components were positively correlated with the comprehensive coagulation index by two statistical methods. Six and eleven monomers were obtained respectively by chromatographic preparation and procurement, and one monomer was eliminated due to preparation difficulty and other reasons. Seven active monomers with direct hemostatic effect and one active monomer with synergistic hemostatic effect were screened through plasma recalcification time, and their combinations were used as candidate HECG for hemostatic effect verification. The results of in vitro experiments showed that plasma recalcification time and in vitro coagulation time were significantly reduced (P < 0.05) in the HECG group, compared to the PNE group. The results of in vivo experiment also indicated that the hemostatic effect of HECG was comparable to that of PNE and PN powder. ConclusionThe composition and efficacy of the HECG of PN were screened and verified using the spectral correlation method and in vivo and in vitro efficacy verification; the HECG included Dencichine, Ginsenoside Rg1, Ginsenoside Rd, Ginsenoside Rh1, Ginsenoside F1, Notoginsenoside R1, Notoginsenoside Ft1 and Notoginsenoside Fe. These results laid a foundation for the quality evaluation of PN and provided a reference for the basic research of pharmacodynamic material basis of other TCMs.