Kyoung Jin Nho

Corosolic acid induces apoptotic cell death in human lung adenocarcinoma A549 cells in vitro

Corosolic acid (CRA), a triterpenoid from medicinal herbs, has been shown to induce apoptosis in several cell lines, with the exception of A549 cells. In this report, we investigated the apoptotic effect and mechanism of CRA in A549 cells. The present study shows that CRA significantly inhibits cell viability in a concentration- and time-dependent manner. Exposure to CRA induces sub-G1 cell cycle arrest and causes apoptotic death in A549 cells. CRA also triggers the activation of caspases and poly(ADP-ribose) polymerase, an effect antagonized by z-vad-fmk. In addition, exposure to CRA leads to a significant increase in the levels of reactive oxygen species (ROS) in A549 cells. Furthermore, exposure to the ROS scavenger N acetylcysteine (NAC)prevents CRA-induced apoptosis, suggesting a role for ROS in CRA-induced apoptosis. ROS are critical regulators of caspase-mediated apoptosis in A549 cells. These results indicate that CRA induces mitochondria-mediated and caspase-dependent apoptosis in A549 cells by altering anti-apoptotic proteins in a ROS-dependent manner.

Topical application of an ethanol extract prepared from Illicium verum suppresses atopic dermatitis in NC/Nga mice

ETHNOPHARMACOLOGICAL RELEVANCE Illicium verum is a traditional herbal medicine with anti-inflammatory properties used in Asia. However, its usefulness in the treatment of allergic diseases remains unclear. This study evaluated the anti-inflammatory and antiallergic effects of I. verum extract (IVE) in a mouse model of atopic dermatitis. MATERIALS AND METHODS We investigated the effects of IVE on compound 48/80-induced histamine release, and phorbol 12-myristate13-acetate and calcium ionophore A23187-stimulated cytokines secretion in MC/9 mast cells. Atopic dermatitis was induced in NC/Nga mice by exposure to extract of house dust mite (Dermatophagoides farinae). After a topical application of IVE on ear and skin lesions, we evaluated the severity of skin symptoms, ear thickness, inflammatory cell infiltration, and serum levels of immunoglobulin E (IgE), histamine, interleukin (IL)-6, and intercellular adhesion molecule (ICAM)-1. In addition, we determined the expression of IL-4, IL-6, tumor necrosis factor (TNF)-α, interferon (IFN)-γ thymus- and activation-regulated chemokine (TARC), regulated on activation, normal T cell expressed and secreted (RANTES), ICAM-1, and vascular cell adhesion molecule (VCAM)-1 in ear tissues. RESULTS IVE inhibited secretion of histamine, IL-4, IL-6, and TNF-α from mast cells in a dose-dependent manner. Topical application of IVE significantly reduced dermatitis scores, ear thickness, and serum levels of IgE, histamine, IL-6, and ICAM-1. Histopathological analysis demonstrated decreased epidermal thickening and dermal infiltration by inflammatory cells. In the ear lesions, IVE treatment reduced expression of IL-4, IL-6, TNF-α, TARC, RANTES, ICAM-1, and VCAM-1, but not IFN-γ. CONCLUSIONS These results indicate that IVE inhibits atopic dermatitis-like skin lesions by suppressing the expression of cytokines, chemokines, and adhesion molecules. These results suggest that IVE may be a potential therapeutic candidate for atopic dermatitis.

Inhibitory effects of Drynaria fortunei extract on house dust mite antigen-induced atopic dermatitis in NC/Nga mice.

ETHNOPHARMACOLOGICAL RELEVANCE Drynaria fortunei (Kunze) J. Sm has been widely used in traditional medicine for the treatment of inflammation, hyperlipidemia, arteriosclerosis, rheumatism, and bone healing. We investigated the anti-inflammatory effects of a 70% ethanol extract of Drynaria fortunei (DFE). MATERIALS AND METHODS We evaluated the anti-inflammatory effects of topically applied DFE on house dust mite Dermatophargoides farinae-induced atopic dermatitis-like skin lesions in NC/Nga mice. RESULTS Treatment of NC/Nga mice with DFE reduced the dermatitis score, ear thickness, and serum levels of IgE, IgG1, and IL-6. Histopathological analyses of ear and skin lesions showed inhibition of the thickening of the epidermis and reduced epidermal/dermal infiltration of inflammatory cells. In ear lesions, mRNA expression levels of IL-4, IL-6, and tumor necrosis factor-α were reduced by DFE treatment. CONCLUSIONS DFE inhibited the development of dermatitis-like skin lesions in NC/Nga mice. These results suggest that DFE may be a therapeutic candidate for the treatment of AD.

Agrimonia pilosa ethanol extract induces apoptotic cell death in HepG2 cells

ETHNOPHARMACOLOGICAL RELEVANCE Agrimonia pilosa (AP) has been used as a traditional herbal medicine for treating various cancers and diseases in Asian countries. MATERIALS AND METHODS Cell viability along with caspase-3/-7, caspase-8 and caspase-9 activity were measured to detect apoptosis. The activity of the apoptotic factors bcl-2, bcl-xl, mcl-1, XIAP, BID, BIK, caspase-3, caspase-9 and PARP were measured by Western blotting. FACS analysis was used to analyze the cell cycle. RESULTS APE inhibited the proliferation of HepG2 cells. Growth inhibition was associated with increased caspase activity and sub-G1 apoptotic fractions. When we measured the affect of APE on intracellular signaling, APE stimulated the apoptotic factors bcl-2, bcl-xl, mcl-1, XIAP, BID, BIK, caspase-3, caspase-9 and PARP in HepG2 cells. CONCLUSIONS The results indicate that APE induces programmed cell death (apoptosis) in HepG2 cells and demonstrates one of the mechanisms underlying the therapeutic effects of the extract reported in previous studies.

An ethyl acetate fraction derived from Houttuynia cordata extract inhibits the production of inflammatory markers by suppressing NF-кB and MAPK activation in lipopolysaccharide-stimulated RAW 264.7 macrophages.

BackgroundHouttuynia cordata Thunb. (Saururaceae) has been used in traditional medicine for treatment of inflammatory diseases. This study evaluated the anti-inflammatory effects of an ethyl acetate fraction derived from a Houttuynia cordata extract (HCE-EA) on the production of inflammatory mediators and the activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.MethodsTo measure the effects of HCE-EA on pro-inflammatory cytokine and inflammatory mediator’s expression in RAW 264.7 cells, we used the following methods: cell viability assay, Griess reagent assay, enzyme-linked immunosorbent assay, real-time polymerase chain reaction and western blotting analysis.ResultsHCE-EA downregulated nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin (IL-6) production in the cells, as well as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, HCE-EA suppressed nuclear translocation of the NF-κB p65 subunit, which correlated with an inhibitory effect on IκBα (nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha) phosphorylation. HCE-EA also attenuated the activation of MAPKs (p38 and JNK).ConclusionsOur results suggest that the anti-inflammatory properties of HCE-EA may stem from the inhibition of pro-inflammatory mediators via suppression of NF-κB and MAPK signaling pathways.

Anti-atherosclerotic effects of Polygonum aviculare L. ethanol extract in ApoE knock-out mice fed a Western diet mediated via the MAPK pathway

ETHNOPHARMACOLOGICAL RELEVANCE Polygonum aviculare L. has been used in traditional Korean medicine to treat obesity and symptoms associated with hypertension. The effectiveness or mechanism of Polygonum aviculare L. ethanol extract (PAE) on atherosclerosis disease has not been examined experimentally. This study investigated the protective effect of PAE in atherosclerotic mice. MATERIALS AND METHODS ApoE KO mice were fed a Western diet (WD) alone or with PAE or a statin for 12 weeks, followed by analysis of bodyweight, serum lipid levels, and blood pressure. Staining of the aorta and adipose tissue, expression levels of adhesion molecules, and the MAPK pathway were also examined. Cell viability, NF-κB activity, and protein levels of adhesion molecules were assessed in vitro. RESULTS ApoE KO mice fed PAE (50 and 100 mg/kg) or statin (10 mg/kg) gained less body weight, and has less adipose tissue and lower serum lipid levels and blood pressures than the WD group. Aorta ICAM-1, VCAM-1, and NF-κB levels were decreased by PAE in a dose-dependent manner, consistent with the in vitro observations. PAE and statin decreased atherosclerotic plaque and adipocyte size versus the WD group. Furthermore, PAE decreased phosphorylation of MAPK pathway components in the aorta of PAE-treated mice, suggesting that PAEs anti-atherosclerotic effects are mediated via a MAPK pathway-dependent mechanism. CONCLUSIONS PAE may protect against the development of atherosclerotic disease. The beneficial effects are associated with lowering bodyweight, serum lipids, blood pressure, adhesion molecular protein levels, atherosclerotic plaque, and adipocyte size, involving the MAPK pathway.

Ethanol Extract of Dianthus chinensis L. Induces Apoptosis in Human Hepatocellular Carcinoma HepG2 Cells In Vitro

Dianthus chinensis L. is used to treat various diseases including cancer; however, the molecular mechanism by which the ethanol extract of Dianthus chinensis L. (EDCL) induces apoptosis is unknown. In this study, the apoptotic effects of EDCL were investigated in human HepG2 hepatocellular carcinoma cells. Treatment with EDCL significantly inhibited cell growth in a concentration- and time-dependent manner by inducing apoptosis. This induction was associated with chromatin condensation, activation of caspases, and cleavage of poly (ADP-ribose) polymerase protein. However, apoptosis induced by EDCL was attenuated by caspase inhibitor, indicating an important role for caspases in EDCL responses. Furthermore, EDCL did not alter the expression of bax in HepG2 cells but did selectively downregulate the expression of bcl-2 and bcl-xl, resulting in an increase in the ratio of bax:bcl-2 and bax:bcl-xl. These results support a mechanism whereby EDCL induces apoptosis through the mitochondrial pathway and caspase activation in HepG2 cells.

Anti-metastatic effects of Rheum Palmatum L. extract in human MDA-MB-231 breast cancer cells.

Rheum palmatum L. (RP) has been widely used in traditional medicine for the treatment of various diseases in Asian countries. The molecular mechanism of its anti-metastasis effect remains elusive. The present study assessed the effect of RP ethanol extract (RPE) on the highly metastatic human MDA-MB-231 breast cancer cells in vitro. At a non-toxic concentration, RPE inhibited migration, motility and invasion in a concentration-dependent manner. To investigate the mechanisms involved, real-time PCR and Western blot analyses were performed. Results showed that RPE down-regulated the levels of extracellular matrix degradation-associated proteins, including MMP-2/-9, uPA and uPAR, and up-regulated PAI-1. In addition, RPE affected NF-κB by degrading IkBα, and affected the mitogen-activated protein kinase signal transduction pathway by depressing the activation of p38, ERK and Akt. These results suggest that RPE has potential anti-metastatic activity and warrants further investigation.

Ampelopsis japonica ethanol extract suppresses migration and invasion in human MDA‑MB‑231 breast cancer cells

Ampelopsis japonica (AJ) is a well‑known traditional oriental herb with anti‑inflammatory and anticancer activities. However, the molecular mechanisms by which AJ inhibits metastasis in breast cancer cells remain to be elucidated. The aim of the present study was to investigate the effects of AJ ethanol extract (EAJ) on highly metastatic human MDA‑MB‑231 breast cancer cells in vitro. AJ was extracted and chemically characterized. Cell proliferation was determined using a CCK‑8 assay and migration was detected using a wound healing motility assay. A Transwell assay was used to evaluate the invasion and metastatic capabilities of the MDA‑MB‑231 cells. In addition, the mRNA expression levels of metalloproteinase (MMP)‑2 and MMP‑9 and tissue inhibitors of metalloproteinases (TIMP)‑1 and TIMP‑2 were evaluated using reverse transcription quantitative polymerase chain reaction in vitro. The results of the present study characterized the signaling cascades that mediated the antimetastatic activity of AJ in the human MDA‑MB‑231 breast cancer cell line. EAJ significantly suppressed the migration and invasion of MDA‑MB‑231 cells in vitro and inhibited the expression of metalloproteinase (MMP)‑2 and MMP‑9. These findings identified the biological activity of EAJ in an in vitro model of cancer metastasis and provided a rationale for further investigation.

A methanol fraction from Chaenomeles sinensis inhibits hepatocellular carcinoma growth in vitro and in vivo

The fruits of Chaenomeles sinensis have numerous therapeutic properties, including anticancer and antiinflammatory activities; however, its antitumor activity and underlying molecular mechanism are poorly understood. The present study evaluated the in vitro and in vivo antitumor activities of a fraction of C. sinensis extract purified on amberlite resin and eluted in 30% methanol (CSAM 30). In vitro, cell viability was assessed using the CCK-8 assay, cell cycle was analyzed by flow cytometry, and apoptosis was measured by Hoechst DNA staining, caspase activity assays and Western blotting. In vivo antitumor efficacy of CSAM 30 was evaluated by oral administration on the human HepG2 hepatocellular carcinoma preclinical xenograft model. In vitro, CSAM 30 inhibited HepG2 cell proliferation and induced apoptosis via activation of caspases, cleavage of poly ADP-ribose polymerase, up-regulation of Bad, and down-regulation of Xlinked inhibitor of apoptosis protein XIAP and bcl-2. In vivo, CSAM 30 inhibited HepG2 tumor growth in a dose-dependent manner without inducing body weight loss. These results demonstrate that CSAM 30 induces apoptosis and has antitumor activity in vivo and in vitro.