Guem-San Lee

Effects of Dictamnus dasycarpus Turcz., root bark on ICAM-1 expression and chemokine productions in vivo and vitro study

ETHNOPHARMACOLOGICAL RELEVANCE The root bark of Dictamnus dasycarpus Turcz., family Rutaceae is a well known anti-inflammatory agent for skin diseases such as eczema, pruritus and urticaria in Eastern countries. MATERIALS AND METHODS We investigated the effects of methanol extract of Dictamnus dasycarpus root bark (MEDD) on Intercellular Adhesion Molecule-1 (ICAM-1) expression, epidermal hyperplasia and immune cell infiltration in 1-fluoro-2,4-dinitrofluorobenzene (DNFB)-induced contact dermatitis (CD) mice. We also investigated its effects on the expression of ICAM-1, binding capacity to THP-1 cells, cytokine and chemokine production, and phosphorylation of NF-κB in human keratinocytes (HaCaT cells). RESULTS Topical application of MEDD effectively inhibited ICAM-1 expression and epidermal hyperplasia in inflamed tissues. MEDD treatment also inhibited immune cell infiltration induced by DNFB. In addition, treatment with MEDD reduced surface expression and total amount of ICAM-1in HaCaT cells and effectively lowered the capacity to bind to THP-1 cells. MEDD also lowered the levels of IL-6, IL-8, monokine induced by gamma interferon (MIG), monocyte chemotactic protein-1 (MCP-1) and regulated on activation, normal T cell expressed and secreted (RANTES). Finally, MEDD treatment prevented activation of the NF-κB pathway induced by TNF-α in HaCaT cells. CONCLUSIONS These data indicate that root bark of Dictamnus dasycarpus has the potential for treatment of inflammatory skin diseases as a complementary or alternative medicine to corticosteroids. In addition, they suggest that the anti-inflammatory effects of Dictamnus dasycarpus on CD are involved in the regulation of ICAM-1 expression and cytokine and chemokine secretion through down-regulation of the NF-κB signaling pathway in keratinocytes.

Comparative study of Korean White Ginseng and Korean Red Ginseng on efficacies of OVA-induced asthma model in mice

Background Korean ginseng is a well-known medicinal herb that has been widely used in traditional medicine to treat various diseases, including asthma. Ginseng can be classified as white ginseng (WG) or red ginseng (RG), according to processing conditions. In this study, the authors compared the efficacies of these two ginseng types in a mouse model of acute asthma. Methods To produce the acute asthma model, BALB/c mice were sensitized with ovalbumin (OVA) and aluminum hydroxide, and then challenged with OVA. WG and RG extracts were administered to mice orally. The influences of WG and RG on airway hyperresponsiveness (AHR), immune cell distributions in bronchoalveolar lavage fluid (BALF), and OVA-specific immunoglobulin E (IgE), IgG1, and IgG2a in serum were investigated. Cytokine production by lymphocytes isolated from peribronchial lymph nodes and histopathological changes was also examined. Results In OVA-sensitized mice, both WG and RG reduced AHR and suppressed immune cell infiltration in bronchoalveolar regions. BALF OVA-specific IgE levels were significantly lower in RG-treated OVA-sensitized mice than in the OVA-sensitized control group. WG and RG also suppressed inflammatory cytokine production by peribronchial lymphocytes. Histopathological findings showed reduced inflammatory cell infiltration and airway remodeling (e.g., epithelial hyperplasia) in WG- and RG-treated OVA mice compared with OVA controls. Conclusion In this study, WG and RG showed antiasthmatic effects in an OVA-sensitized mouse model, and the efficacies of RG were found to be better than those of WG.

Effects of Schisandra chinensis Turcz. fruit on contact dermatitis induced by dinitrofluorobenzene in mice

Schisandra chinensis Turcz. fruit is widely used to treat skin diseases. The aim of this study was to determine the anti-inflammatory effects of the methanol extract of S. chinensis (MESC) on 1-fluoro-2,4-dinitrofluorobenzene (DNFB)-induced contact dermatitis (CD) in mice. The effects of MESC on ear thickness and weight, histopathological changes, immune cell filtration and cytokine production were investigated in DNFB-induced CD mice. Topical application of MESC effectively inhibited ear swelling (30 or 300 μg on the left ear, P<0.001; 30 μg on the right ear, P<0.001). MESC also inhibited hyperplasia, spongiosis (100 μg/ear, P<0.05 and 300 μg/ear, P<0.001, respectively) and immune cell infiltration (100 μg/ear, P<0.05; 300 μg/ear, P<0.001) induced by DNFB. In addition, MESC suppressed increases in tumor necrosis factor (TNF)-α levels (100 or 300 μg/ear, P<0.05), interferon (INF)-γ (30 μg/ear, P<0.05; 100 μg/ear, P<0.01; 300 μg/ear, P<0.001), interleukin (IL)-6 (300 μg/ear, P<0.05) and monocyte chemoattractant protein (MCP)-1 (30 μg/ear, P<0.05; 100 μg/ear, P<0.01; 300 μg/ear, P<0.001). These results suggest that the anti-inflammatory effects of MESC are mediated by the reduced production of TNF-α, IFN-γ, IL-6 and MCP-1, and that MESC has potential use for the treatment of inflammatory skin diseases.

Influence of herbal combinations on the extraction efficiencies of chemical compounds from Cinnamomum cassia, Paeonia lactiflora, and Glycyrrhiza uralensis, the herbal components of Gyeji-tang, evaluated by HPLC method

During decoction process, the ingredients of herbal formula interact with each other, such that therapeutic properties and chemical extraction characteristics are altered. The crude drugs, Cinnamomum cassia (CC), Paeonia lactiflora (PL), and Glycyrrhiza uralensis (GU), are the main herbal constituents of Gyeji-tang, a traditional herbal formula. To evaluate the chemical interaction between CC, PL, and GU during the course of decoction, quantification of 16 marker compounds in the herbal decoction, performed using a Box-Behnken experimental design, was carried out by HPLC-diode array detection using validated method. Correlations between the amounts of marker compounds from CC, PL, and GU were assessed by multiple regression analysis. The results obtained showed that amounts of single herb marker compounds significantly changed (usually decreased) by decoction in the presence of other herbs and that these changes depended on the chemical natures of the markers and the herbal medicines present. Results also demonstrated that the extraction efficiencies of marker compounds increased when the proportion of the herb containing them was increased and decreased in proportion to amounts of herbs added. In conclusion, chemical interactions between compositional herbal medicines may occur when herbs are co-decocted. This study provides insight of understanding the herbal interactions in herbal formulae.

Suppression of TPA-induced cancer cell invasion by Peucedanum japonicum Thunb. extract through the inhibition of PKCα/NF-κB-dependent MMP-9 expression in MCF-7 cells.

Metastatic cancers spread from their site of origin (the primary site) to other parts of the body. Matrix metalloproteinase-9 (MMP-9), which degrades the extracellular matrix, is important in metastatic cancers as it plays a major role in cancer cell invasion. The present study examined the inhibitory effect of an ethanol extract of Peucedanum japonicum Thunb. (PJT) on MMP-9 expression and the invasion of MCF-7 breast cancer cells induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). Western blot analysis, gelatin zymography, and reverse transcription-quantitative PCR revealed that PJT significantly suppressed MMP-9 expression and activation in a dose-dependent manner. Furthermore, PJT attenuated TPA-induced nuclear translocation and the transcriptional activation of nuclear factor (NF)-κB. The results indicated that the PJT-mediated inhibition of TPA-induced MMP-9 expression and cell invasion involved the suppression of the PKCα/NF-κB pathway in MCF-7 cells. Thus, the inhibition of MMP-9 expression by PJT may have potential value as a therapy for restricting the invasiveness of breast cancer.

Nardostachys chinensis induces granulocytic differentiation with the suppression of cell growth through p27Kip1 protein-related G0/G1 phase arrest in human promyelocytic leukemic cells

Abstract Context: Nardostachys chinensis Batalin (Valerianaceae) has been used in Korean traditional medicine to elicit stomachic and sedative effects. However, the anti-leukemic activities of N. chinensis have not been well examined. Objective: To investigate the effect of N. chinensis on differentiation and proliferation in the human promyelocytic leukemic HL-60 cells. Materials and methods: The dried roots and stems of N. chiensis are extracted using hot water and then freeze-dried. The yield of extract was 12.82% (w/w). The HL-60 cells were treated with 25–200 μg/ml of N. chinensis for 72 h or 100 μg/ml of N. chinensis for 24–72 h. Results: Nardostachys chinensis significantly inhibited cell viability dose dependently with an IC50 of 100 μg/ml in HL-60 cells. Nardostachys chinensis induced differentiation of the cells as measured by reduction activity of NBT and expression of CD11b but not of CD14 as analyzed by flow cytometry, which indicates a differentiation toward the granulocytic lineage. Nardostachys chinensis also induced growth inhibition through G0/G1 phase arrest in the cell cycle of HL-60 cells. Among the G0/G1 phase in the cell cycle-related protein, the expression of cyclin-dependent kinase (CDK) inhibitor p27Kip1 was increased in N. chinensis-treated HL-60 cells, whereas the expression levels of CDK2, CDK4, CDK6, cyclin D1, cyclin D3, cyclin E, and cyclin A were decreased. Interestingly, N. chinensis markedly enhanced the binding of p27Kip1 with CDK2 and CDK6. Discussion and conclusion: This study demonstrated that N. chinensis is capable of inducing cellular differentiation and growth inhibition through p27Kip1 protein-related G0/G1 phase arrest in HL-60 cells.

Anti-inflammatory effects of Cryptotympana atrata Fabricius slough shed on contact dermatitis induced by dinitrofluorobenzene in mice.

Background: The slough shed of Cryptotympana atrata Fabricius is widely used to treat skin diseases in China, Japan, and Korea. Objective: To investigate the anti-inflammatory effects of C. atrata on contact dermatitis. Materials and Methods: We investigated the effects of C. atrata methanol extract (MECA) on ear swelling, histophathological changes and cytokine production in 1-fluoro-2,4-dinitrofluorobenzene (DNFB)-induced contact dermatitis (CD) mice. Results: Topical application of MECA effectively inhibited enlargement of ear swelling (30 and 100 μ/ear, P < 0.05; 300 μg/ear, P < 0.01). MECA treatment also inhibited hyperplasia, spongiosis (100 and 300 μg/ear, P < 0.001), and immune cell infiltration (30 μg/ear, P < 0.05; 100 and 300 μg/ear, P < 0.001) induced by DNFB. In addition, treatment with MECA suppressed the increase in the levels of TNF-α (P < 0.05), IFN-g (3, 100 μg/ear, P < 0.05; 300 μg/ear, P < 0.01), and IL-6 (100 μg/ear, P < 0.05; 300 μg/ear, P < 0.01) production. Conclusion: These data suggest that MECA has the potential for use in the treatment of inflammatory skin diseases, including CD. Moreover, the results presented herein indicate that anti-inflammatory actions of MECA are mediated by decreasing production of TNF-α, IFN-γ, and IL-6 in inflamed tissues.

Anti-cancer effects of Kochia scoparia fruit in human breast cancer cells.

Background: The fruit of Kochia scoparia Scharder is widely used as a medicinal ingredient for the treatment of dysuria and skin diseases in China, Japan and Korea. Especially, K. scoparia had been used for breast masses and chest and flank pain. Objective: To investigate the anti-cancer effect of K. scoparia on breast cancer. Materials and Methods: We investigated the anti-cancer effects of K. scoparia, methanol extract (MEKS) in vitro. We examined the effects of MEKS on the proliferation rate, cell cycle arrest, reactive oxygen species (ROS) generation and activation of apoptosis-associated proteins in MDA-MB-231, human breast cancer cells. Results: MTT assay results demonstrated that MEKS decreased the proliferation rates of MDA-MB-231 cells in a dose-dependent manner with an IC50 value of 36.2 μg/ml. MEKS at 25 μg/ml significantly increased the sub-G1 DNA contents of MDA-MB-231 cells to 44.7%, versus untreated cells. In addition, MEKS induced apoptosis by increasing the levels of apoptosis-associated proteins such as cleaved caspase 3, cleaved caspase 8, cleaved caspase 9 and cleaved Poly (ADP-ribose) polymerase (PARP). Conclusion: These results suggest that MEKS inhibits cell proliferation and induces apoptosis in breast cancer cells and that MEKS may have potential chemotherapeutic value for the treatment of human breast cancer.

Crotonis Fructus and Its Constituent, Croton Oil, Stimulate Lipolysis in OP9 Adipocytes

Introduction. Crotonis fructus (CF) is the mature fruit of Croton tiglium L. and has been used for the treatment of gastrointestinal disturbance in Asia. It is well known that the main component of CF is croton oil (CO). The present study is to investigate the effects of CF extracts (CFE) and CO on lipolysis in OP9 adipocytes. Methods. Glycerol release to the culture supernatants was used as a marker of adipocyte lipolysis. Results. Treatment with various concentrations of CFE and CO stimulates glycerol release in a dose-dependent manner. The increase in glycerol release by CFE is more potent than isoproterenol, which is a β-adrenergic agonist as a positive control in our system. The increased lipolysis by CFE and CO was accompanied by an increase of phosphorylated hormone sensitive lipase (pHSL) but not nonphosphorylated HSL protein and mRNA. Pretreatment with H89, which is a protein kinase A inhibitor, significantly abolished the CFE- and CO-induced glycerol release in OP9 adipocytes. These results suggest that CFE and CO may be a candidate for the development of a lipolysis-stimulating agent in adipocytes.

Optimal processing conditions of Boswellia carteri Birdw. using response surface methodology

Background: Boswellia carteri Bridw. is being widely used for its anti-inflammatory properties, as well as for wound healing, antimicrobial, and immunomodulatory properties, and boswellic acids (BAs) are considered to be the main active constituents. Objectives: To investigate optimal conditions of stir-baking process for the resin of B. carteri with vinegar of using response surface methodology (RSM). Materials and Methods: The concentration of acetic acid, heating temperature, and heating time were set as influential factors, and the yields of chemical compounds were the response values which were optimally designed by a Box–Behnken design. The amounts of 11-keto-β-boswellic acid (KBA) and α-boswellic acid (αBA) in B. carteri resin were quantified using high-performance liquid chromatography analysis. Results: Maximum amounts of KBA and αBA in B. carteri resin were obtained using 6% acetic acid for 10 min at 90°C in preliminary test. Two factor interactions, such as acetic acid concentration–heating temperature and heating temperature–heating time, were significantly observed by multiple regression analysis. Optimal processing conditions from RSM were 5.83% for acetic acid concentration, 9.56 min for heating time, and 89.87°C for heating temperature. Under the modified conditions, the experimental value of the response was 11.25 mg/g, which was similar to the predicted value. Conclusions: The results suggest that the optimal conditions for the stir-baking process of B. carteri resin were determined by RSM, which was reliable and applicable to practical processing of herbal medicine. Abbreviations used: BAs: Boswellic acids; KBA: 11 keto β boswellic acid; αBA: α boswellic acid; BBD: Box–Behnken design; RSM: Response surface method; HPLC: High performance liquid chromatography; LOD: Limits of determination; LOQ: Limits of quantification; RSD: Relative standard deviation; ANOVA: Analysis of variance.