Ginsenoside Rh2 Research Articles

Improvement of 1,3-propanediol production in Klebsiella sp. by co-expressing Vitreoscilla hemoglobin and formate dehydrogenase genes

An efficient and flexible protocol was developed for simultaneous detection of the ginsenosides Rg1, Re, Rf, Rh1, and Rg2 in ginseng extract. In the analysis of white ginseng that contains no ginsenoside Rf, separation of the remaining ginsenosides was achieved within just 10 min under isocratic chromatographic conditions. For the analysis of red ginseng that contains ginsenoside Rf as a characteristic constituent, the gradient elution conditions were optimized. The method is based on high-pressure liquid chromatography employing a Shiseido UG 80 Capcell Pak NH2(4.6 mm I.D. × 250 mm, 5 μm)column and isocratic elution using acetonitrile (A) and water(B) in a ratio of 76: 24 (v/v), The optimal gradient elution conditions are as follows: 0–3 min, 89% A, 3–25 min, 89–84% A, 25–30 min, 84–82% A, 30–35 min, 82-76% A, then returning to 89% solvent A in 5 min. The flow rate was 0.80 mL min−1. The column temperature was set at 25℃ and the detection wavelength was at 203 nm. The working concentration ranges for ginsenoside Re, Rh1, Rg2, Rg1, and Rf were 0.23−1450.0 mg• L−1, 0.05−1130.0 mg• L−1, 0.11−687.0 mg• L−1, 0.051−1325 mg• L−1, and 0.55−800.0 mg• L−1, respectively. The method was validated for linearity, precision, and accuracy. And the further confirmation of five ginsenosides was conducted by QTOF-MS. Analysis of raw extracts of ginseng, white ginseng, and red ginseng for the five components showed satisfactory recovery.

Anti-Inflammatory Effect of Ginsenoside Rh2-Mix on Lipopolysaccharide-Stimulated RAW 264.7 Murine Macrophage Cells.

Ginsenoside Rh2, a protopanaxadiol saponin from ginseng, has been reported to have strong anti-inflammatory activity. However, the concentration of ginsenoside Rh2 is very low (>0.001%) in the total ginseng extracted, which is not enough for production despite its high pharmacological effects. Thus, in this study, we evaluated the anti-inflammatory effect of ginsenoside Rh2-mix (GRh2-mix) on lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage cells. From the high-performance liquid chromatography analysis, it was confirmed that the GRh2-mix was mainly composed of 20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3. The LPS-stimulated RAW 264.7 cells were treated with different concentrations of GRh2-mix (100, 200, 400, 500 μg/mL). The cell counting kit-8 assay showed that the GRh2-mix treatment increased cell proliferation in LPS-stimulated RAW 264.7 murine macrophage cells. The GRh2-mix inhibited nitric oxide production in a dose-dependent manner, suggesting an anti-inflammatory effect. Furthermore, reverse transcription polymerase chain reaction and Western blot results also indicated that the GRh2-mix suppressed inflammatory genes such as iNOS, TNF-α, COX-2, IL-1β, IL-6, and NF-κB. In summary, these results suggest that the GRh2-mix exhibits anti-inflammatory activity via the downregulation of the NF-κB pathway and has high efficiency with a simple production procedure.

Biological Evaluation and Docking Studies of Synthetic Oleanane-type Triterpenoids.

Saponins are potential wide-spectrum antitumor drugs, and copper(I) catalyzed azide–alkyne 1,3-dipolar cycloaddition is a suitable approach to synthesizing saponin-like compounds by regioselective glycosylation of the C2/C3 hydroxyl and C28 carboxylic groups of triterpene aglycones maslinic acid (MA) and oleanolic acid (OA). Biological studies on the T-84 human colon carcinoma cell line support the role of the hydroxyl groups at C2/C3, the influence of the aglycone, and the bulky nature of the substituents in C28. OA bearing a α-d-mannose moiety at C28 (compound 18) focused our interest because the estimated inhibitory concentration 50 was similar to that reported for ginsenoside Rh2 against colon cancer cells and it inhibits the G1–S phase transition affecting the cell viability and apoptosis. Considering that triterpenoids from natural sources have been identified as inhibitors of nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) signaling, docking studies were conducted to evaluate whether NF-κB may be a potential target. Results are consistent with the biological study and predict a similar binding mode of MA and compound 18 to the p52 subunit from NF-κB but not for OA. The fact that the binding site is shared by the NF-κB inhibitor 6,6-dimethyl-2-(phenylimino)-6,7-dihydrobenzo[d][1,3]oxathiol-4(5H)-one supports the result and points to NF-κB as a potential target of both MA and compound 18.

Method development for the certification of a ginsenoside calibration solution via liquid chromatography with absorbance and mass spectrometric detection

The research presented here describes the development of two analytical methods for use in the certification of a ginsenoside calibration solution Standard Reference Material (SRM) 3389 consisting of seven ginsenosides: Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd. The new methods utilized the liquid chromatographic (LC) separation of ginsenoside mixtures with absorbance detection (UV) and mass spectrometry (MS). Ginsenosides Rb3, Rg2, Rg3, Rh1, and Rh2 were evaluated for use as internal standards for LC/MS measurements. The 12 ginsenosides were baseline resolved by gradient elution LC/UV, with an initial mobile phase composition of 22% acetonitrile and 78% water, flow rate of 0.7 mL/min, and column temperature of 25 °C. The work presented here includes a detailed investigation into the optimization of the chromatographic conditions to minimize measurement biases that result from unresolved constituents. Temperature and mobile phase composition are known to play a significant role in column selectivity; however, flow rate is expected to influence primarily the separation efficiency and detection sensitivity. In the current study, column selectivity changed with changes in flow rate and the relative retention of ginsenoside Rg2 and Rh1 changed as the flow rate increased from 0.6 mL/min to 1.0 mL/min.

Ginsenoside Rh2 Ameliorates Lipopolysaccharide-Induced Acute Lung Injury by Regulating the TLR4/PI3K/Akt/mTOR, Raf-1/MEK/ERK, and Keap1/Nrf2/HO-1 Signaling Pathways in Mice.

The anti-inflammatory effect of ginsenoside Rh2 (GRh2) has labeled it as one of the most important ginsenosides. The purpose of this study was to identify the anti-inflammatory and antioxidant effects of GRh2 using a lipopolysaccharide (LPS) challenge lung-injury animal model. GRh2 reduced LPS-induced proinflammatory mediator nitric oxide (NO), tumor necrosis factor-alpha, interleukin (IL)-1β, and anti-inflammatory cytokines (IL-4, IL-6, and IL-10) production in lung tissues. GRh2 treatment decreased the histological alterations in the lung tissues and bronchoalveolar lavage fluid (BALF) protein content; total cell number also reduced in LPS-induced lung injury in mice. Moreover, GRh2 blocked iNOS, COX-2, the phosphorylation of IκB-α, ERK, JNK, p38, Raf-1, and MEK protein expression, which corresponds with the growth of HO-1, Nrf-2, catalase, SOD, and GPx expression in LPS-induced lung injury. An in vivo experimental study suggested that GRh2 has anti-inflammatory effects, and has potential therapeutic efficacy in major anterior segment lung diseases.

Evaluation of toxicity of functionalized graphene oxide with ginsenoside Rh2, lysine and arginine on blood cancer cells (K562), red blood cells, blood coagulation and cardiovascular tissue: In vitro and in vivo studies

Abstract In this study, blood- and cardio-toxicity of Rh2–treated graphene oxide (GO-Rh2), lysine-treated graphene oxide (GO-Lys), arginine-treated GO (GO-Arg), Rh2–treated GO-Lys (GO-Lys-Rh2) and Rh2–treated GO-Arg (GO-Arg-Rh2) were evaluated. Two concentrations of each nanostructures (200 and 1000 µg/ml) was injected to rats. After 2 weeks, the effects of agents were evaluated on heart tissue by histopathological assays. Cytotoxicity of all designed nanostructures was investigated on blood cancer cells (K562) by MTT assay. Toxicity of designed nanostructures was also investigated on Red Blood Cells (RBCs), Prothrombin Time (PT) and Partial Thromboplastin Time (PTT). The results demonstrated increase of anticancer activity for GO-Arg-Rh2 and GO-Lys-Rh2 in comparison with free Rh2 and GO. GO, GO-Rh2, GO-Lys, GO-Arg, GO-Lys-Rh2, and GO-Arg-Rh2 had 50% hemolysis at concentrations 250, 360, 420, 435, 500, and 575 µg/ml, respectively. GO led to RBCs aggregation and morphological change at 5–100 µg/ml, but other functionalized nanostructures did not show these changes. All nanostructures had slight effect on intrinsic and extrinsic coagulation system, especially on PTT. GO-Arg-Rh2 and GO-Lys-Rh2 had lower effect on blood coagulation system in comparison with other examined nanosystems. Besides, GO-Rh2, GO-Arg-Rh2, and GO-Lys-Rh2 had lowest toxicity on heart tissue than other synthesized nanostructures. Functionalization of GO with Arg, Lys, and especially, Rh2 led to decrease the destruction of heart tissue. So, modified GO with Rh2 and basic amino acids may be a potential and promising strategy to enhance the therapeutic index for GO because of the reduction of side effects on normal cells.

Hepatoprotective effect of ultrasonicated ginseng berry extract on a rat mild bile duct ligation model

BackgroundThe Panax ginseng berry extract (GBE) is well known to have an antidiabetic effect. The aim of this study is to evaluate and investigate the protective effect of ultrasonication-processed P. ginseng berry extract (UGBE) compared with GBE on liver fibrosis induced by mild bile duct ligation (MBDL) model in rats. After ultrasonication process, the composition ratio of ginsenoside in GBE was changed. The component ratio of ginsenosides Rh1, Rh4, Rg2, Rg3, Rk1, Rk3, and F4 in the extract was elevated. MethodsIn this study, the protective effect of the newly developed UGBE was evaluated on hepatotoxicity and neuronal damage in MBDL model. Silymarin (150 mg/kg) was used for positive control. UGBE (100 mg/kg, 250 mg/kg, 500 mg/kg), GBE (250 mg/kg), and silymarin (150 mg/kg) were orally administered for 6 weeks after MBDL surgery. ResultsThe MBDL surgery induced severe hepatotoxicity that leads to liver inflammation in rats. Also, the serum ammonia level was increased by MBDL surgery. However, the liver dysfunction of MBDL surgery–operated rats was attenuated by UGBE treatment via myeloid differentiation factor 88-dependent Toll-like receptor 4 signaling pathways. ConclusionUGBE has a protective effect on liver fibrosis induced by MBDL in rats through inhibition of the TLR4 signaling pathway in liver.

MicroRNA-146a-5p enhances ginsenoside Rh2-induced anti-proliferation and the apoptosis of the human liver cancer cell line HepG2.

Liver cancer is one of the leading causes of malignancy-associated mortality worldwide and its clinical therapy remains very challenging. Ginsenoside Rh2 (Rh2) has been reported to have antitumor effects on some types of cancer, including liver cancer. However, its regulatory mechanism has not been extensively evaluated. In the present study, Rh2 increased the expression of microRNA (miR)-200b-5p, miR-224-3p and miR-146a-5p, and decreased the expression of miR-26b-3p and miR-29a-5p. Of the three upregulated miRs, miR-146a-5p exhibited the highest fold elevation. In accordance with a previous study, Rh2 effectively inhibited the survival of liver cancer cells in vitro and in a mouse model. In addition, it was observed that Rh2 markedly promoted liver cancer apoptosis and inhibited colony formation. Cell apoptosis and the inhibition of cell survival as well as colony formation induced by Rh2 were enhanced and weakened by miR-146a-5p overexpression and inhibition, respectively. The results of the present study provide further evidence of the antitumor effect of Rh2 in liver cancer and also demonstrate that this effect may be mediated via the regulation of miR-146a-5p expression in the liver cancer cell line HepG2. The results indicated that miR-146a-5p may be a promising regulatory factor in Rh2-mediated effects in liver cancer.

Transformation of ginsenoside Rh4 and its aglycone from the total saponins of stems and leaves of Panax ginseng by Aspergillus tubingensis

Rare ginsenoside Rh4 and its aglycone are present in low levels in Panax ginseng. This study focused on achieving ginsenoside Rh4 and its aglycone by microbial transformation of the total saponins of stems and leaves of P. ginseng (TSSLG). A total of 78 fungal strains isolated from the rhizosphere soil of P. ginseng, and one of them, Aspergillus tubingensis, was found to be capable of biotransformation of TSSLG to ginsenoside Rh4 and its aglycone with high efficiency, especially the content of ginsenoside Rh4 aglycone was enhanced 100 folds over than that in the original TSSLG. Compared with ginsenoside Rg3 (named ShenYi capsule as antitumor drug in China), ginsenoside Rh4 aglycone exhibited stronger antitumor activity against human cancer cell lines SGC-7901, KB-A-1 and HT-1080. The possible biotransformation pathways of ginsenoside Rh4 and its aglycone were speculated according to the chemical structures. The anticancer binding-target, myeloid cell leukemia 1 protein (MCL-1), was explored using molecular docking analysis and the possible combination mechanism was discussed.

Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2

BackgroundTumor-associated macrophages (TAMs) play a critical role in modulating the tumor microenvironment and promote tumor metastases. Our studies have demonstrated that ginsenoside Rh2 (G-Rh2), a monomeric compound extracted from ginseng, is a promising anti-tumor agent in lung cancer cells. However, it remains unclear whetherG-Rh2 can modulate the differentiation of TAMs and its interaction with tumor microenvironment. In this study, we investigated how G-Rh2 regulates the phenotype of macrophages and affects the migration of non-small cell lung cancer (NSCLC) cells.MethodsMurine macrophage-like RAW264.7 cells and human THP-1 monocyte were differentiated into M1 and M2 subsets of macrophages with different cytokines combination, which were further identified by flow cytometry with specific biomarkers. M2 macrophages were sorted out to co-culture with NSCLC cell lines, A549 and H1299. Wound healing assay was performed to examine the cell migration. Expression levels of matrix metalloproteinases 2 and 9 (MMP-2, − 9) and vascular endothelial growth factor-C (VEGF-C) were measured by RT-qPCR and western blot, and the release of VEGF in the supernatant was measured by a VEGF ELISA kit. Finally, modulation of TAMs phenotype and VEGF expression by G-Rh2 was examined in vivo.ResultsWe demonstrated that M2 subset of macrophages alternatively differentiated from RAW264.7 or THP-1cells promote migration of NSCLC cells. Further examinations revealed that NSCLC significantly increased the release of VEGF to the media and elevated the expression levels of VEGF at mRNA and protein levels after being co-cultured with M2 macrophages. Similar alterations in MMP-2 and MMP-9 were observed in NSCLC after being co-cultured. Of note,G-Rh2 had a potential to effectively convert M2 phenotype to M1 subset of macrophages. Importantly, G-Rh2 had a preference to decrease the expression levels of VEGF, MMP2, and MMP9 in co-cultured lung cancer cells, over than those in lung cancer cells without co-culturing. Consistently, G-Rh2 reduced M2 macrophage marker CD206 and VEGF expression levels in vivo.ConclusionsAll of these results suggested that M2 subset macrophages drive lung cancer cells with more aggressive phenotypes. G-Rh2 has a potential to convert TAMs from M2 subset to M1 in the microenvironment and prevents lung cancer cell migration, suggesting the therapeutic effects of G-Rh2onlung cancer.

Membrane cholesterol delays cellular apoptosis induced by ginsenoside Rh2, a steroid saponin

Saponins exhibit several biological and pharmacological activities, such as antibacterial, anti-inflammatory and anticancer effects. Many studies attribute their activities to their interactions with cholesterol. In this study, we focus on the steroid saponin ginsenoside Rh2, one of the active principles of Panax ginseng root. Some evidence suggests that lipid rafts, defined as nanodomains enriched in cholesterol and sphingolipids, could be involved in the Rh2-induced apoptosis. However, the role of membrane lipids, especially cholesterol, in this process is still poorly understood. Here, we demonstrate that (i) A549, THP-1 and U937 cells are all susceptible to the Rh2-induced apoptosis but to a differential extent and (ii) the cytotoxic effect inversely correlates with the cell membrane cholesterol content. Upon cholesterol depletion via methyl-β-cyclodextrin, those three cells lines become more sensitive to Rh2-induced apoptosis. Then, focusing on the cholesterol-auxotroph U937 cell line, we showed that Rh2 alters plasma membrane fluidity by compacting the hydrophobic core of lipid bilayer (DPH anisotropy) and relaxing the interfacial packaging of the polar head of phospholipids (TMA-DPH anisotropy). The treatment with Rh2 conducts to the dephosphorylation of Akt and the activation of the intrinsic pathway of apoptosis (loss of mitochondrial membrane potential, caspase-9 and -3 activation). All these features are induced faster in cholesterol-depleted cells, which could be explained by faster cell accumulation of Rh2 in these conditions. This work is the first reporting that membrane cholesterol could delay the activity of ginsenoside Rh2, renewing the idea that saponin cytotoxicity is ascribed to an interaction with membrane cholesterol.

Corilagin from longan seed: Identification, quantification, and synergistic cytotoxicity on SKOv3ip and hey cells with ginsenoside Rh2 and 5-fluorouracil

Corilagin content from different parts of longan (Dimocarpus longan Lour.) was determined by ultra performance liquid chromatography (UPLC) method. Additionally, the potential synergistic effects of corilagin + ginsenoside Rh2 (Rh2), and corilagin + 5-fluorouracil (5-FU) on ovarian cancer cells, and cancer-preventing activities, including inhibition of tyrosinase, properties of antioxidant and nitrite-scavenging, and blocking of nitrosamine synthesis were investigated. The results showed the content of corilagin from different parts of longan varied widely, while corilagin content in longan seed was high with a value of 542.15 ± 10.30 μg/g. Then the corilagin from longan seed was chosen for further study, since longan seed was easily obtained from by-product of longan fruit processing with low cost. Furthermore, the combinations of corilagin + Rh2, and corilagin + 5-FU showed an increased synergistic cytotoxicity on SKOv3ip and Hey cells. Moreover, corilagin inhibited exhibited effects of inhibiting tyrosinase, antioxidation, scavenging nitrite and blocking nitrosamine synthesis.

The ginsenoside PPD exerts anti-endometriosis effects by suppressing estrogen receptor-mediated inhibition of endometrial stromal cell autophagy and NK cell cytotoxicity

Endometriosis (EMS) is an estrogen-dependent gynecological disease with a low autophagy level of ectopic endometrial stromal cells (eESCs). Impaired NK cell cytotoxic activity is involved in the clearance obstruction of the ectopic endometrial tissue in the abdominopelvic cavity. Protopanaxadiol (PPD) and protopanaxatriol (PPT) are two metabolites of ginsenosides, which have profound biological functions, such as anti-cancer activities. However, the role and mechanism of ginsenosides and metabolites in endometriosis are completely unknown. Here, we found that the compounds PPD, PPT, ginsenoside-Rg3 (G-Rg3), ginsenoside-Rh2 (G-Rh2), and esculentoside A (EsA) led to significant decreases in the viability of eESCs, particularly PPD (IC50 = 30.64 µM). In vitro and in vivo experiments showed that PPD promoted the expression of progesterone receptor (PR) and downregulated the expression of estrogen receptor α (ERα) in eESCs. Treatment with PPD obviously induced the autophagy of eESCs and reversed the inhibitory effect of estrogen on eESC autophagy. In addition, eESCs pretreated with PPD enhanced the cytotoxic activity of NK cells in response to eESCs. PPD decreased the numbers and suppressed the growth of ectopic lesions in a mouse EMS model. These results suggest that PPD plays a role in anti-EMS activation, possibly by restricting estrogen-mediated autophagy regulation and enhancing the cytotoxicity of NK cells. This result provides a scientific basis for potential therapeutic strategies to treat EMS by PPD or further structural modification.

Retracted: Ginsenoside Rh2 inhibits proliferation and migration of medulloblastoma Daoy by down‐regulation of microRNA‐31

This study aimed to investigate the effects of ginsenoside Rh2 on proliferation, apoptosis, and migration of the human medulloblastoma cell line Daoy, as well as to explore the potential mechanisms of the effects. The human medulloblastoma cell line Daoy was cultured in vitro and treated with or without ginsenoside Rh2. CCK-8 assay was performed to investigate the effect of Rh2 on cell survival using a cell counting Kit-8. Cell proliferation was assessed by BrdU assay. Cell apoptosis was determined using flow cytometry analysis. Cell migration was detected using a modified two-chamber migration assay. MiR-31 mimic and the NC control were transfected into Daoy cells and detected by qRT-PCR. The expression of Wnt3a, Wnt5a, and β-catein was detected by Western blot analysis. Rh2 efficiently suppressed the proliferation and migration, and promoted the apoptosis of Daoy cells. Additionally, Rh2 could down-regulate miR-31. miR-31 overexpression reversed the effects of Rh2 on proliferation, apoptosis and migration of Daoy cells, and activated the Wnt/β-catein signaling pathways in Daoy cells. Rh2 could inhibit the proliferation and migration, and induce apoptosis of Daoy medulloblastoma cells through down-regulation of miR-31 to inactivate the Wnt/β-catein signaling pathway. Therefore, Rh2 may have a utility in clinical applications for the treatment of medulloblastoma.

Pseudo-Ginsenoside Rh2 induces A549 cells apoptosis via the Ras/Raf/ERK/p53 pathway.

Ginsenoside Rh2, a major effective constituent of ginseng, has been suggested to have a pro-apoptotic effect in a variety of cancer cells. Pseudo-Ginsenside-Rh2 (pseudo-G-Rh2) is a novel derivative of ginsenoside Rh2. The aim of the present study was to evaluate the effect of pseudo-G-Rh2 on the apoptosis of lung adenocarcinoma A549 cells. The cytotoxicity of pseudo-G-Rh2 on A549 cells was evaluated using an MTT assay. Apoptosis was detected using DAPI staining and flow cytometry. The expression of apoptosis associated proteins was identified by western blot analysis. The results demonstrated that pseudo-G-Rh2 inhibits the proliferation of A549 cells in a dose-dependent manner. DAPI staining revealed topical morphological changes in apoptotic bodies following pseudo-G-Rh2 treatment. Flow cytometric analysis revealed that the percentage of Annexin V-fluorescein isothiocyanate-positive cells, which are apoptotic, increased with pseudo-G-Rh2 treatment in a dose-dependent manner. Furthermore, treatment with pseudo-G-Rh2 increased the level of reactive oxygen species in A549 cells as well as the activation of caspase-9, caspase-3 and poly ADP-ribose polymerase. Pseudo-G-Rh2 treatment was observed to induce mitochondrial membrane potential loss. Furthermore, the results of western blotting revealed that B-cell lymphoma 2 (Bcl-2) expression was significantly decreased while Bcl-2-associated X protein expression was significantly upregulated in A549 cells with pseudo-G-Rh2 treatment. Pseudo-G-Rh2-induced apoptosis was accompanied by sustained phosphorylation of Ras, Raf, extracellular signal-regulated kinase (ERK) and p53. In conclusion, the results of the present study suggest that pseudo-G-Rh2 induces mitochondrial apoptosis in A549 cells and is responsible for excessive activation of the Ras/Raf/ERK/p53 pathway.

The anti-tumor effect of ginsenoside Rh4 in MCF-7 breast cancer cells in vitro and in vivo

Breast cancer is a tremendous threat to humans in many countries, and thus we need to find safe and effective drugs for treatment. Ginsenoside Rh4 has been reported to be present in processed ginseng. However, few studies have focused on its anti-tumor activity. In this study, we investigated the inhibitory effects of ginsenoside Rh4 on MCF-7 breast cancer cells and the pathways that promote apoptosis in vitro. To study the effect of ginsenoside Rh4 in vivo, xenograft models were randomly divided into 3 groups (the control group, 10 mg/kg/d Rh4, 20 mg/kg/d Rh4, n = 10 per group), the ginsenoside Rh4 injection method was i.p. The results showed that ginsenoside Rh4 effectively inhibited proliferation, arrested the cell cycle in S phase and induced apoptosis in MCF-7 cells by flow cytometry. Morphological changes caused by ginsenoside Rh4-induced apoptosis were also observed by Hoechst 33342 staining. Western-blot analyses indicated that the apoptosis-inducing effects of ginsenoside Rh4 were associated with the external pathway by decreasing Bcl-2, increasing Bax, and activating caspase-8, -3 and PARP. Moreover, ginsenoside Rh4 significantly inhibited the growth of MCF-7 tumor cells in vivo. These results suggested that ginsenoside Rh4 could be a potentially effective anti-tumor drug for breast cancer.

The reversal effect of Ginsenoside Rh2 on drug resistance in human colorectal carcinoma cells and its mechanism.

Recent studies hint that Ginsenoside is involved in cancer prevention and treatment. In this study, we investigated the effect of Ginsenoside Rh2 ondrug resistanceinhumancolorectalcarcinoma (CRC)cells and its mechanism. The resistance reversion effect of Ginsenoside Rh2 in CRC cells was analyzed using CCK-8 assay. After treating with Ginsenoside Rh2, the cell cycle distribution and cellular apoptosis were analyzed by flow cytometry, cell migration was determined by transwell migration assay, the expression of drug-resistance genes and proteins were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Ginsenoside Rh2 could enhance the cytotoxicity of 5-FU in drug-resistant CRC cells (LoVo/5-FU and HCT-8/5-FU). Treatment with Ginsenoside Rh2 could result in an increase of cell numbers in G0/G1 phase accompanied with a decrease in S-phase, and induced cellular apoptosis in drug-resistant CRC cells. In addition, the migration process and EMT process of drug-resistant CRC cells were suppressed by treatment of Ginsenoside Rh2. Compared to control group, expression of drug-resistance genes, such as MRP1, MDR1, LRP and GST, were negatively correlated to Ginsenoside Rh2. All these results indicated that Ginsenoside Rh2 could effectively reverse drug resistancein humancolorectalcarcinoma cell and its mechanism involved the prevention of cellular proliferation and migration, the promotion of cellular apoptosis and the alteration of drug-resistance genes, which suggested that Ginsenoside Rh2 may act as a promising candidate for drug resistance in humancolorectalcarcinoma chemotherapy.

COMPARATIVE STUDY OF ECHINOCHROME A, OXYGENATED CAROTENOIDS, GINSENOSIDE Rh2, LUTEOLIN DISULFATE AND METFORMIN AS A MEAN TO POTENTIATE ANTITUMOR EFFECT OF DOXORUBICIN

This work concerned a comparative study of potentiating effects produced by some drugs of natural origin, as well as metformin, upon antitumor activity of doxorubicin. An increased therapeutic efficacy of doxorubicin used in combination with echinochrome A, oxycarotenoids, ginsenoside Rh2, and metformin was tested in the in vivo Ehrlich ascites carcinoma model. The negative effect of luteolin disulfate (10 mg/kg body weight) upon efficacy of antitumor therapy was revealed. When studying the effect of these substances on the level of intracellular ROS in primary cultures of murine spleen cells, using a selective fluorescent indicator (2’,7’-dichlorofluorescein-diacetate), we did not find a positive correlation between induction of excessive ROS synthesis and potentiation of doxorubicin action with the compounds studied. Some of the studied agents showed a positive correlation between interferon-inducing activity, as well as a negative correlation between increased IL-1 content, and higher antitumor effect of doxorubicin. The obtained results suggest expedience of further studies of antitumor effect potentiation using preparations of echinochrome, ginsenoside Rh2, a mixture of oxygenated carotenoids and metformin.

Ginsenoside Rh2 reverses sleep deprivation-induced cognitive deficit in mice

Sleep deprivation (SD) negatively caused cognitive deficit, which was associated with oxidative stress induced damage. Ginsenoside Rh2 had the ability to protect against damage caused by reactive oxygen species in vitro, showing antioxidant property. Therefore, it was hypothesized that Ginsenoside Rh2 could prevent SD-induced cognitive deficit via its antioxidant properties. In this study, the effect of Ginsenoside Rh2 on memory impairment induced by sleep deprivation was investigated. The mice were sleep deprived continuously for 14 days using our self-made Sleep Interruption Apparatus (SIA). Ginsenoside Rh2 was administered intraperitoneally at two doses (20 and 40 μmol/kg) for 20 days. Thereafter, behavioral studies were conducted to test the learning and memory ability using object location recognition (OLR) experiment and passive avoidance (PA) test. Additionally, the oxidative stress parameters in the serum and the brain tissues (cortex and hippocampus) were assessed, including the superoxide dismutase (SOD) enzyme activity, the total antioxidant reactivity (TAR), the malondialdehyde (MDA) level, the glutathione (GSH) level, and the lipid peroxidation (LPO) content. The results revealed that SD impaired both spatial and non-spatial memory (P < 0.05). Treatment with Ginsenoside Rh2 at both doses prevented memory impairment induced by SD. Moreover, Ginsenoside Rh2 normalized the reduction of SOD and TAR activities in the serum (P < 0.01) and the decrease of GSH content in both the cortex and hippocampus (P < 0.05) induced by SD. Furthermore, Ginsenoside Rh2 significantly decreased the MDA level in the serum (P < 0.05) and the LPO content in both the cortex and hippocampus (P < 0.05) compared to SD group. In conclusion, sleep deprivation impaired both spatial and non-spatial memory and Ginsenoside Rh2 reversed this impairment, probably by preventing the oxidative stress damage in the body, including the serum and brain during sleep deprivation.

20(S)-Ginsenoside Rh2 Induce the Apoptosis and Autophagy in U937 and K562 Cells

Acute myeloid leukemia (AML) and Chronic myelogenous leukemia (CML) are common leukemia in adults. 20(S)-GRh2 is an important bioactive substance that is present in Panax ginseng. However, there are no investigations that deal with the comparison of apoptosis, the occurrence of autophagy, and the relationship between apoptosis and autophagy after being treated with 20(S)-GRh2 in AML and CML. In this study, we explored the effect of 20(S)-GRh2 on the AML and CML (U937 and K562). Fluorescence microscopy, CCK-8, Quantitative realtime PCR, Western blot, transmission electron microscopy (TEM), and flow cytometric analysis were used to detect the occurrence of cell proliferation inhibition, apoptosis, and autophagy. By using the above methods, it was determined that apoptosis induced by 20(S)-GRh2 was more obvious in K562 than U937 cells and 20(S)-GRh2 could generate autophagy in K562 and U937 cells. When pretreated by a specific inhibitor of autophagy, (3-methyladenine), the 20(S)-GRh2-induced apoptosis was enhanced, which indicated that 20(S)-GRh2-induced autophagy may protect U937 and K562 cells from undergoing apoptotic cell death. On the other hand, pretreated by an apoptosis suppressor (Z-VAD-FMK), it greatly induced the autophagy and partially prevented 20(S)-GRh2 induced apoptosis. This phenomenon indicated that 20(S)-GRh2-induced autophagy may serve as a survival mechanism and apoptosis and autophagy could act as partners to induce cell death in a cooperative manner. These findings may provide a rationale for future clinical application by using 20(S)-GRh2 combined autophagy inhibitors for AML and CML.