The Anti-Tumor Effect and Underlying Apoptotic Mechanism of Ginsenoside Rk1 and Rg5 in Human Liver Cancer Cells

Ginsenoside Rk1 and Rg5 are minor ginseng saponins that have received more attention recently because of their high oral bioavailability. Each of them can effectively inhibit the survival and proliferation of human liver cancer cells, but the underlying mechanism remains largely unknown. Network pharmacology and bioinformatics analysis demonstrated that G-Rk1 and G-Rg5 yielded 142 potential targets, and shared 44 putative targets associated with hepatocellular carcinoma. Enrichment analysis of the overlapped genes showed that G-Rk1 and G-Rg5 may induce apoptosis of liver cancer cells through inhibition of mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signal pathways. Methyl thiazolyl tetrazolium (MTT) assay was used to confirm the inhibition of cell viability with G-Rk1 or G-Rg5 in highly metastatic human cancer MHCC-97H cells. We evaluated the apoptosis of MHCC-97H cells by using flow cytometry and 4′,6-diamidino-2-phenylindole (DAPI) staining. The translocation of Bax/Bak led to the depolarization of mitochondrial membrane potential and release of cytochrome c and Smac. A sequential activation of caspase-9 and caspase-3 and the cleavage of poly(ADP-ribose) polymerase (PARP) were observed after that. The levels of anti-apoptotic proteins were decreased after treatment of G-Rk1 or G-Rg5 in MHCC-97H cells. Taken together, G-Rk1 and G-Rg5 promoted the endogenous apoptotic pathway in MHCC-97H cells by targeting and regulating some critical liver cancer related genes that are involved in the signal pathways associated with cell survival and proliferation.     

The effectiveness of borneol on pharmacokinetics changes of four ginsenosides in Shexiang Baoxin Pill in vivo

Shexiang Baoxin Pill (SBP) is a traditional Chinese medicine, widely used for cardiovascular diseases in the clinic. Ginsenosides are important effective components in SBP, but their pharmacokinetic characteristics are still not known. In this paper, we studied the pharmacokinetics of ginsenoside Rb1, Rc, Re and Rg1 in SBP and investigated the effect of borneol on the pharmacokinetic characteristic of ginsenosides based on an Agilent G6410A triple quadrupole LC/MS system. Results showed that the pharmacokinetic parameters of ginsenoside Rb1, Rc, Re and Rg1 in rat plasma after oral administration of SBP are significantly different with oral administration of SBP without Borneolum Syntheticum. Plasma pharmacokinetic profiles after oral administration of ginsenoside Rb1, Rc, Re, Rg1 and co‐administration with borneol at three different ratios (10:1, 1:1 and 1:10 ginsenoside vs borneol, w/w) were also determined. It was demonstrated that borneol can elevate the plasma concentration of ginsenosides after co‐admininstration. Copyright © 2013 John Wiley & Sons, Ltd.     

Metabolite profiling of ginsenoside Rg1 after oral administration in rat

The goal of this study is to investigate the biotransformation of ginsenoside Rg₁ in vivo. A highly sensitive and specific LC‐MS/MS method was developed and used for metabolite identification in rat feces and urine after oral administration of ginsenoside Rg₁. Four metabolites of Rg₁ were detected in rat feces and three metabolites of Rg₁ were detected in rat urine. Deglycosylation and oxygenation were found to be the major metabolic pathways of ginsenoside Rg₁ after oral administration in rat. Except for the reported metabolites Rh₁ and protopanaxatriol, mono‐oxygenated Rg₁ and mono‐oxygenated protopanaxatriol were detected for the first time after oral administration of Rg₁. The in vivo metabolite profiling of ginsenoside Rg₁ in rat was proposed. Viewed collectively, Rg₁ was metabolized to mono‐oxygenated Rg₁, Rh₁, protopanaxatriol and the secondary metabolite mono‐oxygenated protopanaxatriol in rat. Copyright © 2014 John Wiley & Sons, Ltd.     

Significant difference in active metabolite levels of ginseng in humans consuming Asian or Western diet: The link with enteric microbiota

After ingestion of ginseng, the bioavailability of its parent compounds is low and enteric microbiota plays an important role in parent compound biotransformation to their metabolites. Diet type can influence the enteric microbiota profile. When human subjects on different diets ingest ginseng, their different gut microbiota profiles may influence the metabolism of ginseng parent compounds. In this study, the effects of different diet type on gut microbiota metabolism of American ginseng saponins were investigated. We recruited six healthy adults who regularly consumed different diet types. These subjects received 7 days' oral American ginseng, and their biological samples were collected for LC‐Q‐TOF‐MS analysis. We observed significant ginsenoside Rb₁ (a major parent compound) and compound K (a major active metabolite) level differences in the samples from the subjects consuming different diets. Subjects on an Asian diet had much higher Rb₁ levels but much lower compound K levels compared with those on a Western diet. Since compound K possesses much better cancer chemoprevention potential, our data suggested that consumers on a Western diet should obtain better cancer prevention effects with American ginseng intake compared with those on an Asian diet. Ginseng compound levels could be enhanced or reduced via gut microbiota manipulation for clinical utility.     

Metabolite profiling of ginsenosides in rat plasma,urine and feces by LC–MS/MS and its application to a pharmacokinetic study after oral administration of Panax ginseng extract

Panax ginseng is widely consumed as a functional food in the form of tea, powder, capsules, among others, and possesses a range of pharmacological activities including adaptogenic, immune‐modulatory, anti‐tumor, anti‐aging and anti‐inflammatory effects. The aim of this study was to identify and quantify the major ginsenosides and their metabolites in rat plasma, urine and feces after administration of P. ginseng extract using LC–MS/MS. We collected rat plasma samples at 0.5, 1, 2, 4, 8, 12, 24 and 48 h, and the amounts of urine and fecal samples accumulated in 24 h. Fourteen major ginsenosides and their metabolites were observed in fecal samples at high levels; however, low levels of 11 ginsenosides were detected in urine samples. The pharmacokinetics of the major ginsenosides and their metabolites was investigated in plasma. The results indicated that the maximum plasma concentration, time to maximum concentration and area under the curve of compound K were significantly greater than those of other ginsenosides. This study thus provides valuable information for drug development and clinical application of P. ginseng.     

高效液相色谱法分离测定人参中的6种人参皂甙

 采用梯度法,以乙腈-水溶液为流动相,用高效液相色谱法分离人参中的6种主要人参皂甙,并采用紫外检测器检测,在203 nm处测定4种人参样品。该方法在25 mg/L～300 mg/L的范围内有良好的线性关系,回收率高于80%。     

Isolation,characterization and phase transformation of new ginsenoside compound k hydrate and methanol solvates

Two new solvates of ginsenoside compound K (nonstoichiometric hydrate/CKH and methanol solvate/CKM) have been discovered and characterized in this paper. They were obtained through cooling crystallization in different solvents, and CKM could be prepared by transformation from CKH as well. The solvates were analysed by Power X‐ray diffraction (PXRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and fourier transform infrared (FTIR) spectroscopy. From the thermal studies, it is shown that the two new products are both solvates with different onset melting points. The PXRD and FTIR data support different crystal structures of them. It also describes the solution‐mediated phase transformation from CKH to CKM with a combination usage of process analytical technology tools. It is shown that the transformation process can be divided into three stages. The results reveal that seeding and low temperature help to accelerate the transformation, but initial solution concentration do little to the transformation kinetics. The kinetics and the rate‐controlling step for the transformation depend on the nucleation of the CKM. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

高效液相色谱-电喷雾质谱联用法测定人参和西洋参的皂苷类成分

利用高效液相色谱（LC—ELSD）与电喷雾质谱（ESI—MS）联用技术对人参和西洋参中的人参皂苷类成分进行了比较研究。通过液相色谱与质谱联用技术获得了相应化合物的分子量信息;利用质谱的源内CID技术获得了相应化合物的结构信息。根据人参皂苷Rf和拟人参皂苷F11两种同分异构体在质谱中的源内CID裂解规律的不同,建立了人参和西洋参药材的LC-ESI-MS简便、准确的鉴别方法。     

电喷雾质谱法研究人参皂苷与溶菌酶的相互作用

用电喷雾质谱法研究了溶菌酶与人参皂苷Rg1和Re的非共价相互作用。非共价复合物的解离常数可以由溶菌酶和复合物的峰高直接计算得到。由各个体系得到的溶菌酶与Rg1和Re的一级解离常数KD, 1是一致的，但是二级解离常数KD, 2的差别较大。由此可以推断出复合物的峰高越强，结果的精密度越好。当固定溶菌酶的浓度不变时，复合物的解离常数会随着人参皂苷的浓度的增加而略微的增大。而当固定人参皂苷的浓度时，复合物的解离常数会随着溶菌酶的浓度的增加而减小。计算结果还证明了人参皂苷Rg1与溶菌酶的结合能力大于Re。     

20(R)和20(S)-人参皂甙Rg_2碳氢NMR信号全指定

20 (R)和 2 0 (S) 人参皂甙 Rg2 属于达玛烷型四环三萜类化合物 .应用 2DNMR技术 :1 H 1 HCOSY、HMQC和HMBC全归属 2 0 (R)和 2 0 (S) 人参皂甙 Rg2 碳和氢质子信号 ,为该类型化合物的结构鉴定提供波谱学依据 .