Shakina Yesmin Simu

Ginsenoside Rg5:Rk1 attenuates TNF-α/IFN-γ-induced production of thymus- and activation-regulated chemokine (TARC/CCL17) and LPS-induced NO production via downregulation of NF-κB/p38 MAPK/STAT1 signaling in human keratinocytes and macrophages

Atopic dermatitis (AD) is a chronic skin disease that affects millions of people worldwide. Keratinocytes and macrophages are two cells types that play a pivotal role in the development of AD. These cells produced different chemokines and cytokines, especially thymus and activation-regulated chemokine (TARC/CCL17) and macrophage-derived chemokine (MDC/CCL22), as well as nitric oxide (NO) through inducible nitric oxide synthase (iNOS) and COX2 in response to stimulation by TNF-α/IFN-γ and lipopolysaccharide (LPS) respectively. These mediators are thought to be crucial regulators of the pathogenesis of AD. Although several natural compounds to treat AD have been studied, the effect of Rg5:Rk1 from Panax ginseng (P. ginseng) on AD has not yet been investigated. In this study, we evaluated the inhibitory effect of Rg5:Rk1 on TNF-α/IFN-γ stimulated keratinocytes (HaCaT cells) and LPS-stimulated macrophages (RAW 264.7 cells). Enzyme-linked immunosorbent assay (ELISA) data showed that pretreatment of HaCaT cells with Rg5:Rk1 significantly reduced the TNF-α/IFN-γ-induced increase in TARC/CCL17 expression in a dose-dependent manner. In addition, Rg5:Rk1 decreased LPS-mediated nitric oxide (NO) and reactive oxygen species (ROS) production in RAW 264.7 cells. A considerable reduction in messenger RNA (mRNA) expression of the aforementioned AD mediators was also observed. Pretreatment with Rg5:Rk1 attenuated the TNF-α/IFN-γ-induced phosphorylation of p38 MAPK, STAT1, and NF-κB/IKKβ in HaCaT cells. Together, these findings suggest that ginsenoside Rg5:Rk1 may have a potential anti-AD effect by suppressing NF-κB/p38 MAPK/STAT1 signaling.

Anticancer activity of silver nanoparticles from Panax ginseng fresh leaves in human cancer cells.

The pharmaceutical role of silver nanoparticles has been increased over the last decades, especially those synthesized through herbal medicinal plants, due to their variety of pharmacological importance. Panax ginseng Meyer (P. ginseng) has been widely used as a therapeutic herbal medicine for a long time in cancer treatment. In this study, the cytotoxic and oxidative effect of a novel silver nanoparticles synthesized from P. ginseng fresh leaves (P.g AgNPs) were evaluated in different human cancer cell lines. In addition, the effect of P.g AgNPs on cell migration, apoptosis and the determination of the mechanism involve was determinate by the use of A549 lung cancer cell line. It was found that P.g AgNPs treatment inhibited cell viability and induced oxidative stress in A549, MCF7 and HepG2 cancer cell lines. Likewise, P.g AgNPs treatment inhibited the epidermal growth factor (EGF)-enhanced migration, as well as decreased the mRNA levels and phosphorylation of EGF receptors in A549 cells. Moreover, P.g AgNPs modified the morphology of the cell nucleus and increase apoptosis percentage; this effect was linked to the stimulation of p38 MAPK/p53 pathway. Taken together, our results showed that P.g AgNPs exhibited anti-cancer activity in A549 and the regulation of EGFR/p38 MAPK/p53 pathway might be the possible mechanism of its anti-activity. Further experiments are suggested to determinate the mechanism by which P.g AgNPs induce cytotoxicity and ROS generation in MCF-7 and HepG2 cells.

Gold nanoparticles synthesized using Panax ginseng leaves suppress inflammatory - mediators production via blockade of NF-κB activation in macrophages.

Abstract In the present study, we report that Gold nanoparticles (AuNPs) synthesized using the leaf extract of Panax ginseng Meyer (P.g AuNPs) exert anti-inflammatory effects through inhibition of downstream NF-κB activation in macrophages. We found that P.g AuNPs reduced the expression of the inflammatory mediators including nitric oxide (NO), prostaglandin E2 (PEG2), interleukin (IL)-6, tumor necrosis factor-α (TNF-α) was attenuated by P.g AuNPs. Furthermore, P.g AuNPs suppressed lipopolysaccharide (LPS)-induced activation of NF-κB signaling pathway via p38 mitogen-activated protein kinase (MAPK) in RAW 264.7 cells. Taken together, our results suggest that P.g AuNPs can be utilized as a novel therapeutic agent for the prevention and cure of inflammation.

Inhibition of Osteoclast Differentiation by Ginsenoside Rg3 in RAW264.7 Cells via RANKL, JNK and p38 MAPK Pathways Through a Modulation of Cathepsin K: An In Silico and In Vitro Study

Various studies have demonstrated that overexpression of cathepsin K (Cat‐K) causes excessive bone loss, which ultimately leads to a variety of bone diseases including osteoporosis. Therefore, inhibition of Cat‐K signifies a potential therapeutic target in osteoporosis treatment. Ginsenoside Rg3 is one of the most promising compound of Panax ginseng Meyer (P. ginseng) with numerous biological activities. Thus, in recent study the inhibitory effect of Rg3 isolated from P. ginseng was investigated in order to impede the osteoclast activity by an in silico approach followed by in vitro study validation using RAW264.7 cells through the investigation of different biological activity prediction such as absorption distribution metabolism and excretion (ADMET) properties against Cat‐K protein. The docking results of our study showed that Rg3 is a non‐toxic compound and may act as a drug‐like molecule. Additionally, the molecular interaction of Rg3 with the active residues of Cat‐K markedly describes its inhibitory effects on osteoclastogenesis. Findings of the present study exhibited that Rg3 significantly reduced receptor activator of nuclear factor kappa B ligand (RANKL)‐induced tartrate‐resistant acid phosphatase (TRAP) activity, pit formation (actin rings), and TRAP‐positive multinucleated cells development in RAW264.7 cells. Furthermore, Rg3 dose‐dependently reduced the mRNA expression levels of osteoclast‐specific markers such as RANK, TRAP, and Cat‐K induced by RANKL through the down regulation of p38, extracellular signal‐regulated kinase, and c‐Jun N‐terminal kinase (JNK) pathways. In conclusion, in silico docking study and in vitro validation together suggested that Rg3 inhibits osteoclastogenesis and reduces bone resorption through the inhibition of Cat‐K. Therefore, Rg3 might be a useful therapeutic agent for the treatment of osteoporosis and proper bone formation. Copyright

Silver nanoparticles from Dendropanax morbifera Léveille inhibit cell migration, induce apoptosis, and increase generation of reactive oxygen species in A549 lung cancer cells.

Green synthesized silver nanoparticles have significant potential in the pharmaceutical field because of their biological functions such as antioxidant and anticancer activities. Novel silver nanoparticles synthesized from Dendropanax morbifera Léveille leaves (D-AgNPs) exhibit antimicrobial activity and reduce the viability of cancer cells without affecting the viability of RAW 264.7 macrophage-like cells. In this study, we evaluated the anticancer effect of D-AgNPs by measuring the levels of reactive oxygen species (ROS) production and toxicity against A549 and HepG2 cell lines. The effect of D-AgNPs on cell migration, induction of apoptosis, and modification of gene and/or protein expression of cancer-related markers was determined using A549 cells. D-AgNPs exhibited cytotoxicity in A549 and HepG2 cell at different concentrations and enhanced the production of ROS in both cell lines. An increase in cell apoptosis and a reduction in cell migration in A549 cells were also observed after D-AgNP treatment. Furthermore, the effect of D-AgNPs in A549 cells was shown to be related to modification of the EGFR/p38 MAPK pathway. Our data provide the first evidence supporting the potential of D-AgNPs as a possible anticancer agent, particularly for the treatment of non-small cell lung carcinoma.

Suppression of MAPKs/NF-κB Activation Induces Intestinal Anti-Inflammatory Action of Ginsenoside Rf in HT-29 and RAW264.7 Cells.

ABSTRACT This study investigated the intestinal anti-inflammatory action of ginsenoside Rf in inflammatory bowel disease (IBD). IBD is a chronic inflammatory disease that affects the intestinal tract. It is associated with elevated levels of various inflammatory mediators, including interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), nitric oxide (NO), and reactive oxygen species (ROS). Ginsenosides, the main active constituents of ginseng, have been reported to exert potent therapeutic effects against diverse diseases. However, ginsenoside Rf treatment for inflammation has not yet been examined. In this study, we evaluated the inhibitory effect of ginsenoside Rf on the inflammatory mediators downstream of p38/NF-kB activation on TNF-α-stimulated intestinal epithelial cells (HT-29) and mouse macrophage cells (RAW264.7). Our results showed that ginsenoside Rf significantly reduced the production of IL-1β, IL-6, TNF-α, NO, and ROS, which are most highly activated in IBD. In addition, ginsenoside Rf significantly suppressed TNF-α/LPS-induced NF-κB transcriptional activity. These results suggest that ginsenoside Rf contains a compound that has potent intestinal anti-inflammatory effects that could be used to treat diseases such as IBD.

Pharmacological importance, characterization and applications of gold and silver nanoparticles synthesized by Panax ginseng fresh leaves

Abstract Previously, we showed the rapid and eco-friendly synthesis of gold and silver nanoparticles within 3 and 45 min by fresh leaves extract of herbal medicinal plant Panax ginseng. In addition, we characterized the nanoparticles in terms of shape, size, morphology and stability by FE-TEM, EDX, elemental mapping, SEAD, XRD and particles size analysis. In addition of this, we showed their antimicrobial, anti-coagulant, and biofilm inhibition activity of nanoparticles. Continuing our previous study, here we highlight the further characterization and biomedical applications of P. ginseng leaf-mediated gold and silver nanoparticles. We characterized the nanoparticles further in terms of active functional group and capping layer, surface charge, and temperature stability. Based on these factors, we explored the nanoparticles for antioxidant efficacy, biocompatibility in HaCaT cells, 3T3-L1 pre-adipocytes cells, for anticancer efficacy in A549 lung cancer and B16BL6 skin melenoma cancer cell lines and for anti-inflammation efficacy in RAW 264.7 cell lines. Based on our findings, we suggest that the P. ginseng-mediated gold nanoparticles have high antioxidant activity and highly biocompatibility in HaCaT cells, 3T3-L1 pre-adipocytes cells, RAW 264.7 cells lines and could be considered for future drug delivery carriers. The silver nanoparticles also showed high potent antioxidant efficacy, additionally it showed high anticancer effect in A549 lung cancer and B16BL6 skin melenoma cancer cell lines as compared to precursor salts. Moreover, both gold and silver nanoparticles have anti-inflammatory efficacies in RAW 264.7 cells. Thus, the study may provide useful insights of P. ginseng leaves extract-mediated biocompatible gold and silver nanoparticles and improving their applicability in designing nanoparticles carrier systems for drug delivery applications.

Gold nanoflowers synthesized using Acanthopanacis cortex extract inhibit inflammatory mediators in LPS-induced RAW264.7 macrophages via NF-κB and AP-1 pathways

We reported the rapid synthesis (<8s) of gold nanoparticles at room temperature using Acanthopanacis cortex extract (A-AuNPs). We characterized the A-AuNPs using several analytical techniques and found that nano-flower type A-AuNPs, which are known to possess a coarse surface with a high surface to volume ratio, conferring these particles with high binding capacity for various biological molecules. After confirming the stability of the nanoparticles, we investigated the anti-inflammatory effect of A-AuNPs in LPS-stimulated RAW264.7 cells. These nanoparticles inhibited LPS-induced iNOS and COX-2 protein as well as gene expression level, along with reduction of NO and PGE2 production. Furthermore, we observed that the A-AuNPs inhibited translocation of NF-κB and AP-1 through phosphorylation of MAPK signaling by western blot analysis. In summary, we synthesized gold nanoflowers in an economical and eco-friendly way using Acanthopanacis cortex extract and the resultant flower-like A-AuNPs had anti-inflammatory activity, highlighting their potential as therapeutic candidates for suppression of inflammatory-mediated diseases.

Caspase-3/MAPK pathways as main regulators of the apoptotic effect of the phyto-mediated synthesized silver nanoparticle from dried stem of Eleutherococcus senticosus in human cancer cells

Siberian ginseng (Eleutherococcus senticosus) was used for the synthesis of an ecofriendly silver nanoparticle (Sg-AgNP), which has exhibited antibacterial, antioxidant effect and lower cytotoxicity to normal cells in comparison to human cancer cells. Although, the potential anticancer activity of Sg-AgNP has not been determined. In this study, two cancer cell lines were used to evaluate the cytotoxicity and apoptotic effect of Sg-AgNP along with the determination of the role of the Caspase-3 / p38 MAPK pathways. Results shown that Sg-AgNP reduced the cell viability of colon cancer cells HT29 and lung cancer cells A549. The cytotoxic effect was higher than the effect exhibited by a commercial silver nanoparticle and Cisplatin. Reactive oxygen species were observed to be superior in both cell lines in the presence of Sg-AgNPs than c-AgNPs and Cisplatin. It was observed an activation of MAPK14 gene and phosphorylation of p38 MAPK protein in both cell lines induced by Sg-AgNPs treatment. Furthermore, induction of morphological changes in the nucleus was done by Sg-AgNPs at 10 μg/mL in both cell lines. On the other hands, the activation of CASP3 gene and Caspase-3 protein was observed in HT29 cells but only at protein level in A549 cells. These results, suggest that Sg-AgNPs anticancer potential activity might be linked to the induction of apoptosis though the generation of ROS by activation of the Caspase-3/p38 MAPK pathway.

Ginsenoside F1 attenuates lipid accumulation and triglycerides content in 3T3-L1 adipocytes with the modulation of reactive oxygen species (ROS) production through PPAR-γ/JAK2 signaling responses

Obesity is the severe health concern worldwide, causing a highest risk of mortality rate every year in different countries. During adipogenesis interactions with peroxisome proliferator-activated receptor gamma are mainly responsible for the development of obesity with regulation of the various transcription factors. It causes the increase of lipid accumulation, triglyceride content as well as reactive oxygen species production in adipocyte differentiation. Current drugs used as anti-obesity associated with several side effects. The present study was therefore conducted, to evaluate the protective and inhibitory effect of ginsenoside F1 from Panax ginseng on lipid accumulation and reactive oxygen species production by an in silico and in vitro study using 3T3-L1 cells. The structures of F1 compound has been obtained from own in-house Panax ginseng saponin data base. In silico molecular docking and drug likeness properties including absorption, distribution, metabolism and excretion, and toxicity and prediction of biological activity were performed in order to assess and investigate the antiobesity activity of F1. Molecular docking study showed that F1 exhibited strong hydrogen binding affinity with Janus activated protein kinase-2 greater than −9.2 Kcal/mol compared to co-crystalized ligand 3kc and nicotinamide (control drug) −8.2 kcal/mol and −4.6 Kcal/mol respectively. In addition, our in vitro results indicated that F1 also showed significant inhibitory effect on the triglyceride content and reactive oxygen species production in adipocyte differentiation. Moreover, F1 significantly reduced the mRNA expression of adipogenesis markers such as peroxisome proliferator-activated receptor gamma, adipocyte fatty acid-binding protein, and Janus activated prot, which are responsible for lipid accumulation. Our findings demonstrated that ginsenoside F1 inhibits lipid accumulation and reactive oxygen species generation by downregulating the expression adipocyte differentiation markers in mature 3T3-L1 cells.