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Ginsenoside Compound K (CK) has anti-cancer and anti-inflammatory pharmacological activities. It has not been isolated from natural ginseng and is mainly prepared by deglycosylation of protopanaxadiol. Compared with the traditional physicochemical preparation methods, the preparation of CK by hydrolysis with protopanaxadiol-type (PPD-type) ginsenoside hydrolases has the advantages of high specificity, environmental-friendliness, high efficiency and high stability. In this review, the PPD-type ginsenoside hydrolases were classified into three categories based on the differences in the glycosyl-linked carbon atoms of the hydrolase action. It was found that most of the hydrolases that could prepare CK were PPD-type ginsenoside hydrolase type Ⅲ. In addition, the applications of hydrolases in the preparation of CK were summarized and evaluated to facilitate large-scale preparation of CK and its development in the food and pharmaceutical industries.
Subject(s)
Ginsenosides/pharmacology , Hydrolases , Sapogenins/chemistryABSTRACT
This study aims to explore the targets of ginsenosides in brain based on drug affinity responsive target stability(DARTS) technology. Specifically, DARTS technology was combined with label-free liquid chromatography tandem mass spectrometry(LC-MS) to screen out the proteins in the brain that might interact with ginsenosides. Based on the screening results, adenylate kinase 1(AK1) was selected for further confirmation. First, the His-AK1 fusion protein was yielded successively through the construction of recombinant prokaryotic expression vector, expression of target protein, and purification of the fusion protein. Biolayer interferometry(BLI) was employed to detect the direct interaction of Rg_1, Re, Rb_1, Rd, Rh_2, F1, Rh_1, compound K(CK), 25-OH-PPD, protopanaxa-diol(PPD), and protopanaxatriol(PPT) with AK1, thereby screening the ginsenoside monomer or sapogenin that had strong direct interaction with the suspected target protein AK1. Then, the BLI was used to further determine the kinetic parameters for the binding of PPD(strongest interaction with AK1) to His-AK1 fusion protein. Finally, molecular docking technology was applied to analyze the binding properties between the two. With DARTS and LC-MS, multiple differential proteins were screened out, and AK1 was selected based on previous research for target verification. Fusion protein His-AK1 was obtained by prokaryotic expression, and the response(nm) of Re, Rg_1, Rd, Rb_1, Rh_1, Rh_2, F1, PPT, PPD, 25-OH-PPD, and CK with His-AK1 was respectively 0.003 1, 0.001 9, 0.042 8, 0.022 2, 0.013 4, 0.037 3, 0.013 9, 0.030 7, 0.140 2, 0.016 0, and 0.040 8. The K_(on), K_(off), and K_D values of PPD and His-AK1 were determined by the BLI as 1.22×10~2 mol~(-1)·L·s~(-1), 1.04×10~(-2) s~(-1), 8.52×10~(-5) mol·L~(-1). According to the molecular docking result, PPD bound to AK1 with the absolute value of the docking score of 3.438, and hydrogen bonds mainly formed between the two. Thus, AK1 is one of the protein action sites of ginsenosides in the brain. The direct interaction between ginsenoside metabolite PPD and AK1 is the strongest.
Subject(s)
Brain/metabolism , Chromatography, Liquid , Ginsenosides , Molecular Docking Simulation , TechnologyABSTRACT
OBJECTIVE ：To est ablish the method for the determination of 20（S）-protopanaxadiol（PPD）concentration in human plasma. METHODS ：Plasma samples were precipitated with acetonitrile and determined by UPLC-MS/MS ，using finandrogen as internal standard. The determination was performed on Waters ACQUITY UPLC HSS T 3 column with mobile phase consisted of 5 mmol/L ammonium bicarbonate aqueous solution-acetonitrile （gradient elution ）at the flow rate of 0.4 mL/min. The column temperature was set at 40 ℃，and sample size was 10 μL. The ion source was electrospray ion source，and negative ion scanning was carried out with multiple reaction monitoring mode . The ion pairs used for quantitative analysis were m/z 459.40→ 375.20（PPD）and m/z 371.30→315.30（internal standard ）. At the same time ，the method was applied to the determination of clinical samples. RESULTS ：The linear range of PPD was 0.25-30.00 ng/mL（r＝0.999 2），and the limit of quantitation was 0.25 ng/mL. RSDs of intra-batch and inter-batch were all lower than 10％，and relative errors （RE）were －14.61％-12.69％. Extraction method and matrix effect did not affect the quantitative determination of PPD. In ginsenoside CK 100 mg group ，ginsenoside CK 200 mg group and ginsenoside CK 300 mg group ，mean cmax of patients with rheumatoid arthritis after oral administration of corresponding drugs were 18.06，30.03，27.00 ng/mL；median tmax were 12.0，6.0，12.0 h；mean AUC 0-t were 622.52，668.15， 1 155.97 ng·h/mL. CONCLUTIONS ：The method for the determination of PPD concentration in human plasma is successfully established. The method is sensitive ，accurate， kq1907011） stable，easy to operate and less plasma consumption. It can be used for the quantitative determination of clinical samples.
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BACKGROUND AND OBJECTIVES: MUC5AC is one of the major secretory mucin genes in the human airway epithelium. MUC5AC expression is increased by a variety of inflammatory mediators. Protopanaxadiol (PPD), one of the major active metabolites in ginseng, is known to have anti-inflammatory, antitumor and antioxidant properties. However, the effects of PPD on mucin secretion of airway epithelial cells still have not been reported. Therefore, the aim of this study is to investigate the effect of PPD on lipopolysaccharide (LPS)-induced MUC5AC expression in human airway epithelial cells. MATERIALS AND METHOD: In the mucin-producing human NCI-H292 airway epithelial cells, the effect of PPD on MUC5AC expression was investigated using reverse transcription-polymerase chain reaction and enzyme immunoassay after treated with LPS. N-acetylcysteine (NAC) as a reactive oxygen species (ROS) scavenger, and apocynin as a nicotinamide adenine dinucleotide phosphate oxidase inhibitor were used to compare the inhibitory effect of PPD on LPS-induced ROS production in human NCI-H292 cells. RESULTS: LPS significantly increased MUC5AC mRNA expression and protein production. LPS also increased ROS production. PPD inhibited LPS-induced MUC5AC mRNA expression and protein production as well as ROS production. In addition, NAC and apocynin inhibited LPS-induced MUC5AC mRNA expression and protein production. CONCLUSION: These results demonstrate that PPD inhibits LPS-induced MUC5AC expression via ROS in human airway epithelial cells and the inhibitory effect of PPD was similar to that of NAC and apocynin. These findings indicate that PPD may be a therapeutic agent for control of mucus secretion and oxidative stress in human airway epithelial cells.
Subject(s)
Humans , Acetylcysteine , Epithelial Cells , Epithelium , Immunoenzyme Techniques , Methods , Mucins , Mucus , NADP , Oxidative Stress , Oxidoreductases , Panax , Reactive Oxygen Species , RNA, MessengerABSTRACT
Protopanaxadiol (PPD) has inhibitory effects on many tumors and receives much attention. However,it has poor water solubility and low utilization, which limits its clinical application. Considering these issues,in this study,we used hollow gold nanoparticles as transport carriers of PPD and synthesized PPD hollow gold nanoparticles (HAuNs). We conducted a number of experiments to investigate the in vitro anti-laryngeal cancer Hep-2 effect of a PPD HAuNs carrier. High performance liquid chromatography(HPLC) was used to detect the sustained release effect of PPD HAuNs. MTT assay was used to detect the inhibitory effect of PPD HAuNs on the proliferation of Hep-2 cells. Effect of PPD HAuNs on Hep-2 cell apoptosis was investigated by flow cytometry. The results of in vitro release showed that PPD HAuNs had sustained release effect. Compared with blank control group,HAuNs group and PPD group,the survival rate of Hep-2 cells in HAuNs-PEG-PPD group decreased more significantly and the apoptosis rate increased more significantly (p<0.01). PPD HAuNs could significantly enhance anti-laryngeal cancer effect of PPD in vitro and promote the apoptosis of tumor cells. It promotes tumor cell apoptosis, and is expected to be a new PPD drug delivery system, further promoting the application of PPD in clinical anti-laryngeal cancer.
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The intracellular calcium ions (Ca) act as second messenger to regulate gene transcription, cell proliferation, migration and death. Accumulating evidences have demonstrated that intracellular Cahomeostasis is altered in cancer cells and the alteration is involved in tumor initiation, angiogenesis, progression and metastasis. Targeting derailed Casignaling for cancer therapy has become an emerging research area. This review summarizes some important Cachannels, transporters and Ca-ATPases, which have been reported to be altered in human cancer patients. It discusses the current research effort toward evaluation of the blockers, inhibitors or regulators for Cachannels/transporters or Ca-ATPase pumps as anti-cancer drugs. This review is also aimed to stimulate interest in, and support for research into the understanding of cellular mechanisms underlying the regulation of Casignaling in different cancer cells, and to search for novel therapies to cure these malignancies by targeting Cachannels or transporters.
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Objective: In order to evaluate the quality of Panax ginseng and its preparation, a simple and accurate HPLC method for determining the contents of 16 ginsenosides from P. ginseng was established. Methods: The chromatographic separation was achieved on a C18 column (150 mm × 4.6 mm, 5 μm) using a mobile phase made up of acetonitrie and water at a flow rate of 1.0 mL/min. The detection wavelength and column temperature were set as 203 nm and 35°C, respectively. Results: Sixteen ginsenosides (Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, F1, Rd, F2, Rg3, protopanaxatriol, compound K, Rh2, and protopanaxadiol) were separated at baseline with good linearity (r ≥ 0.9990). The recovery rates were 95%-102% (RSD < 2%). Conclusion: The method is simple, fast, accurate, and could be applied to the quality control of P. ginseng and its preparation.
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Objective: Ginsenoside Rg3 was prepared by converting protopanaxadiol (PPD)-type saponins using lemon juice as the catalyst. Methods: Preparation of ginsenoside Rg3 was optimized by response surface method (RSM) based on a four-factor and five-level central composite design. Results: The optimal yield of ginsenoside Rg3 was predicted to be 75.98% in the combination of the factors (PPD-type saponins concentration of 23.6 mg/mL, lemon juice concentration of 97.6% at 85.7 °C for 130.0 min) through the canonical analysis and ridge analysis with maximum responses. Under the optimum conditions, the actual yield of ginsenoside Rg3 was 75.57%. Conclusion: RSM is effective to optimize the preparation of ginsenoside Rg3 by lemon-catalyzed PPD-type saponins. The achievement for the preparation of ginsenoside Rg3 would promote the development of health-care production. © 2013 Tianjin Press of Chinese Herbal Medicines.
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Objective: To prepare 20(R)-ginsenoside Rg3 selectively and to provide the theory basis for the preparation. Methods: 20(R)-Ginsenoside Rg3 was prepared by hydrolyzing protopanaxadiol (PPD)-type saponins using tartaric acid as the catalyst. The preparation condition was optimized by one-factor experiment and orthogonal test, and the reaction products were analyzed by HPLC. Results: The optimization result of orthogonal test showed that when PPD-type saponins (10 mg/mL) were hydrolyzed by tartaric acid (1.5 mol/L) at 110°C for 2.5 h, all the ginsenosides Rb1, Rc, Rb2, Rb3, and Rd were converted, the yields of 20(R)-ginsenoside Rg3 was 50.15%, and the diastereomer excess percentage (de%) was 93.12%. Conclusion: This method is simple, low-cost, and suitable for the mass production, which is very important to promote the study on the pharmacological activities of 20(R)-ginsenoside Rg3.
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Aim To study the anti-tumor effect of 20(S)-Protopanaxadiol(PPD)at different concentration on human liver cell line SMMC-7721 in vivo and in vitro.Methods The subcutaneous transplantable tumor model of human liver cancer in nude mice was established and the anti-tumor effect was calculated.Cell growth rate was determined with MTT assay.The apoptosis was analyzed by FITC-AnnexinⅤ/PI and Hoechst33342 staining method,and the activity of Caspase-3 was detected.Results In vivo,PPD could obviously inhibit the growth of transplanted tumor.In vitro,PPD induced inhibition of human liver cancer SMMC-7721 cells was time-dependent and dose-dependent.The apoptotic body was observed by Hoechst33342 staining.PPD could induce cell apoptosis of SMMC-7721,and the increase of Caspase-3 activity was observed in each PPD group.Conclusion PPD could inhibit the growth of human liver cancer SMMC-7721 cells in vivo and in vitro by up-regulating the activity of Caspase-3 and inducing the cell apoptosis.
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Objective To study the chemical components from the pericarps of Juglans mandshurica with anti-tumor activity.Methods It showed that pericarps of J.mandshurica had a good performance in treating cancer and low toxicity from CHCl3 and EtOAc portions by a lot of pharmacological experiments and modern clinical application in the past.Compounds were isolated by chromatography on silica gel columns and AB-8 macroporous resin.On the basis of spectral evidences including 1D-NMR and 2D-NMR,the structures of these compounds were determined.Results These compounds from CHCl3 portion were identified as 20(S)-protopanaxadiol-3-one(1),dammara-20,24-dien-3?-ol(2),galeon(3),juglanin A(4),2?,3?,23-trihydroxy olean-12-en-28-oic acid(5),2?,3?,23-trihydroxy urs-12-en-28-oic acid(6),oleanolic acid(7),ursolic acid(8).Conclusion Compound 1 is a new natural product.Compounds 2 and 5-8 are isolated firstly from the plants of Juglans L.which are due to pentacyclic triterpane.Compound 3 is isolated from the pericarps of J.mandshurica for the first time.
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Object To observe the effect of 20S-protopanaxadiol saponins from Panax quinquefolium (PPDS) on total cholesterol, lipoprotein cholesterol metabolism and antioxidative activity in experimental hyperlipidemia rats. Methods The total cholesterol (TC), lipoprotein cholesterol, and lipid peroxidation (LPO) contents, prostaglandin I 2 (PGI 2), thromboxane A 2 (TXA 2) levels, superoxide dismutase (SOD) activity and blood viscosity were measured in hyperlipidemia rats which have been given PPDS 25, 50, 100 mg/(kg?d) by ip, continuously for 12 days. In addition, fat accumulation in liver was observed. Results Triglyceride (TG), TC, low density lipoprotein cholesterol (LDL-c) in serum, TXA 2 in plasma, LPO in serum and liver, and blood viscosity were decreased significantly; and high density lipoprotein cholesterol (HDL-c) in serum, PGI 2 in plasma, and SOD in serum and liver were significantly increased by given PPDS [50, 100 mg/(kg?d)] in experimental hyperlipidemia rats. Moreover, PPDS can decrease TC/HDL-c and LDL-c/HDL-c ratio, increase PGI 2/TXA 2 ratio, and inhibit fat accumulation in liver. Conclusion PPDS could inhibit arteriosclerosis by improving cholesterol and lipoprotein-cholesterol metabolism, suppressing LPO, and increasing the activity of SOD.