Mi-Young Jeong

Microarray analysis of gene expression profile by treatment of Cinnamomi Ramulus in lipopolysaccharide-stimulated BV-2 cells

It has been reported that components of Cinnamomi Ramulus (CR) demonstrate an anti-inflammatory effect by inhibiting the expression of inducible nitric oxide synthesis (iNOS) and cyclooxygenase-2 (COX-2) and by suppressing nitric oxide (NO) production in the central nervous system (CNS) as well as in the periphery. In this study, microarray analysis was performed to investigate the effect of CR on the gene expression and associated pathways of lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Microglia plays an important role in the processes of several inflammation-mediated neurodegenerative diseases. Activated microglia can produce various pro-inflammatory cytokines, chemokines, and toxic mediators, which may initiate or amplify the inflammatory responses in the CNS. In the present study, the negative control group was cultured in normal medium, the positive control group was activated with 1 microg/ml of LPS, and the CR group was previously treated with 10 microg/ml of CR before LPS stimulation. With the cutoff value of 1.5-fold change in the expression, 341 genes including pro-inflammatory cytokines, chemokines, and transcription factors were found to be up-regulated in LPS-stimulated BV-2 cells (Supplemental Table 2). CR reduced the LPS-induced up-regulation of such inflammatory genes as Ccl5, Cd80, Cxcl10, Grin2a, Ifi203, Ifit1, Il1alpha, Il6, Lilrb3, Nos2 (iNOS), Rab2b, Rsad2 and Vpreb1. This resulted in a full list of 38 and 37 annotated genes whose expression is up- and down-regulated by CR respectively (Supplemental Table 3). RT-PCR analysis showed that the expression of LPS-induced TNF-alpha, IL-1beta, IL-6, and iNOS mRNAs were attenuated in the presence of the CR extract. The results imply that CR has the anti-inflammatory effect of down-regulating the expression of various genes related to inflammatory responses in LPS-stimulated BV-2 cells, and that CR could be a candidate for the prevention or therapeutic treatment of inflammation-mediated neurodegenerative diseases.

Moutan Cortex Radicis inhibits inflammatory changes of gene expression in lipopolysaccharide-stimulated gingival fibroblasts

Moutan Cortex Radicis (MCR), the root bark of Paeonia suffruticosa Andrews (Paeoniaceae), is found in the traditional Chinese medicinal formulae which were used to treat periodontal diseases. This study investigated the changes in gene expression by MCR treatment when stimulated with lipopolysaccharide (LPS) in cultured human gingival fibroblasts (HGFs). A genome-wide expression GeneChip was used for the gene array analysis, and real-time reverse transcription polymerase chain reaction (RT-PCR) analysis was also performed to confirm the gene expression. It was shown that 42 of the 643 genes up-regulated by LPS, when compared to the control, were down-regulated by the MCR treatment. Of these 42 genes, the inflammation and immune response-related genes were especially noted, which indicates that MCR inhibits the induction of inflammation by LPS stimulation. In addition, 33 of the 519 genes down-regulated by LPS, when compared to the control, were up-regulated by the MCR treatment. The expression patterns of some representative genes by real-time RT-PCR correlated with those of the genes shown in the microarray. In addition, the MCR extract contained paeonol and paeoniflorin, which are known to have the anti-inflammatory effect as the major phenolic components of MCR. This study showed that the MCR extract could comprehensively inhibit a wide variety of activations of inflammation-related genes, which may be due to paeonol and paeoniflorin. It is, thus, suggested that MCR may be applied to alleviate the inflammation of periodontal diseases.

Rubi Fructus (Rubus coreanus) Inhibits Differentiation to Adipocytes in 3T3-L1 Cells

Rubi Fructus (RF) is known to exert several pharmacological effects including antitumor, antioxidant, and anti-inflammatory activities. However, its antiobesity effect has not been reported yet. This study was focused on the antidifferentiation effect of RF extract on 3T3-L1 preadipocytes. When 3T3-L1 preadipocytes were differentiating into adipocytes, 10–100 μg/mL of RF was added. Next, the lipid contents were quantified by Oil Red O staining. RF significantly reduced lipid accumulation and downregulated the expression of peroxisome proliferator-activated receptor γ (PPARγ), CCAAT0-enhancer-binding proteins α (C/EBPα), adipocyte fatty acid-binding protein 2 (aP2), resistin, and adiponectin in ways that were concentration dependent. Moreover, RF markedly upregulated liver kinase B1 and AMP-activated protein kinase (AMPK). Interestingly, pretreatment with AMPKα siRNA and RF downregulated the expression of PPARγ and C/EBPα protein as well as the adipocyte differentiation. Our study shows that RF is capable of inhibiting the differentiation of 3T3-L1 adipocytes through the modulation of PPARγ, C/EBPα, and AMPK, suggesting that it has a potential for therapeutic application in the treatment or prevention of obesity.

Interaction of Veratrum nigrum with Panax ginseng against Obesity: A Sang-ban Relationship

Obesity has become a major health threat in developed countries. However, current medications for obesity are limited because of their adverse effects. Interest in natural products for the treatment of obesity is thus rapidly growing. Korean Medicine (KM) is characterized by the wide use of herbal formulas. However, the combination rule of herbal formulas in KM lacks experimental evidence. According to Shennongs Classic of Materia Medica, the earliest book of herbal medicine, Veratrum nigrum (VN) has antagonistic features against Panax ginseng (PG), and the PG-VN pair is strictly forbidden. In this study, we have shown the effects of PG, VN, and their combination on obesity in high-fat (HF) diet-induced obese mice and in 3T3-L1 cells. PG, VN, and PG-VN combination significantly reduced weight gain and the fat pad weight in HF diet-induced obese mice. They also significantly decreased lipid accumulation and the expressions of two major adipogenesis factors, PPARγ and C/EBPα, in 3T3-L1 cells. In addition, the PG-VN combination had synergistic effects compared with the mixture of extracts of PG and VN on inhibition of PPARγ and C/EBPα expressions at lower doses. These results indicate a new potential anti-obese pharmacotherapy and also provide scientific evidence supporting the usage of herbal combinations instead of mixtures in KM.

Arctigenin Inhibits Adipogenesis by Inducing AMPK Activation and Reduces Weight Gain in High-Fat Diet-Induced Obese Mice.

Although arctigenin (ARC) has been reported to have some pharmacological effects such as anti‐inflammation, anti‐cancer, and antioxidant, there have been no reports on the anti‐obesity effect of ARC. The aim of this study is to investigate whether ARC has an anti‐obesity effect and mediates the AMP‐activated protein kinase (AMPK) pathway. We investigated the anti‐adipogenic effect of ARC using 3T3‐L1 pre‐adipocytes and human adipose tissue‐derived mesenchymal stem cells (hAMSCs). In high‐fat diet (HFD)‐induced obese mice, whether ARC can inhibit weight gain was investigated. We found that ARC reduced weight gain, fat pad weight, and triglycerides in HFD‐induced obese mice. ARC also inhibited the expression of peroxisome proliferator‐activated receptor gamma (PPARγ) and CCAAT/enhancer‐binding protein alpha (C/EBPα) in in vitro and in vivo. Furthermore, ARC induced the AMPK activation resulting in down‐modulation of adipogenesis‐related factors including PPARγ, C/EBPα, fatty acid synthase, adipocyte fatty acid‐binding protein, and lipoprotein lipase. This study demonstrates that ARC can reduce key adipogenic factors by activating the AMPK in vitro and in vivo and suggests a therapeutic implication of ARC for obesity treatment. J. Cell. Biochem. 117: 2067–2077, 2016.

Inhibitory effect of quercetin on colorectal lung metastasis through inducing apoptosis, and suppression of metastatic ability

BACKGROUND Quercetin is a major dietary flavonoid found in a various fruits, vegetables, and grains. Although the inhibitory effects of quercetin have previously been observed in several types of cancer cells, the anti-metastatic effect of quercetin on colorectal metastasis has not been determined. PURPOSE This study investigated whether quercetin exhibits inhibitory effect on colorectal lung metastasis. STUDY DESIGN The effects of quercetin on cell viability, mitogen-activated protein kinases (MAPKs) activation, migration, invasion, epithelial-mesenchymal transition (EMT) and lung metastasis were investigated. METHODS We investigated the effect of quercetin on metastatic colon cancer cells using WST assay, Annexin V assay, real-time RT-PCR, western blot analysis and gelatin zymography. The anti-metastatic effect of quercetin in vivo was confirmed in a colorectal lung metastasis model. RESULTS Quercetin inhibited the cell viability of colon 26 (CT26) and colon 38 (MC38) cells and induced apoptosis through the MAPKs pathway in CT26 cells. Expression of EMT markers, such as E-, N-cadherin, β-catenin, and snail, were regulated by non-toxic concentrations of quercetin. Moreover, the migration and invasion abilities of CT26 cells were inhibited by quercetin through expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) regulation. Quercetin markedly decreased lung metastasis of CT26 cells in an experimental in vivo metastasis model. CONCLUSION In conclusion, this study demonstrates for the first time that quercetin can inhibit the survival and metastatic ability of CT26 cells, and it can subsequently suppress colorectal lung metastasis in the mouse model. These results indicate that quercetin may be a potent therapeutic agent for the treatment of metastatic colorectal cancer.

A combined extract of Cinnamomi Ramulus, Anemarrhenae Rhizoma and Alpiniae Officinari Rhizoma suppresses production of nitric oxide by inhibiting NF‐κB activation in RAW 264.7 cells

An herbal mixture prepared with Cinnamomi Ramulus, Anemarrhenae Rhizoma and Alpiniae Officinari Rhizoma (CAA) is used in oriental medicine for treating several ailments. The purpose of this study was to determine the mechanisms by which CAA elicits an antiinflammatory effect on nitric oxide (NO) production in the mouse macrophage cell line RAW 264.7 cells. The results indicated that lipopolysaccharide (LPS)‐induced NO production was inhibited by CAA in a dose‐dependent manner. Western blotting and RT‐PCR analysis demonstrated that CAA decreased LPS‐induced inducible nitric oxide synthase (iNOS) protein and gene expression in RAW 264.7 cells. Furthermore, CAA inhibited the LPS‐induced DNA binding activity of nuclear factor‐kappa B (NF‐κB) and this effect was mediated through inhibiting the degradation of inhibitory factor‐κBα (IκBα). Therefore, the results demonstrate that CAA inhibits LPS‐induced production of NO and expression of iNOS by blocking NF‐κB activation. CAA might be a potential therapeutic candidate for treating inflammatory diseases such as arthritis. Copyright

Protective changes of inflammation-related gene expression by the leaves of Eriobotrya japonica in the LPS-stimulated human gingival fibroblast: microarray analysis.

ETHNOPHARMACOLOGICAL RELEVANCE The leaf of Eriobotrya japonica (Ej) has been used for a long time as an oriental medicine to treat pulmonary inflammatory diseases. This study investigated the gene expression changes by lipopolysaccharide (LPS) stimulation in the cultured human gingival fibroblast and the anti-inflammatory changes of the genes by the leaves of Ej when challenged with LPS using a microarray chip. MATERIALS AND METHODS A whole transcript genechip (Affymetrix genechip human gene 1.0 ST array) was used. The expression patterns of the significant genes were confirmed by real-time RT-PCR analysis. RESULTS The gene array analysis showed that 60 of the 325 genes up-regulated by the LPS when compared to the control were down-regulated by the Ej treatment. Of these 60 genes, the inflammation- and immune response-related genes were especially noted, which indicates that Ej inhibits the induction of the inflammation through LPS stimulation. In addition, 78 of the 158 genes down-regulated by the LPS when compared to the control were up-regulated by the Ej treatment. The regulatory patterns of the representative genes in the real-time RT-PCR correlated with those of the genes shown in the microarray. The Ej extract also inhibited the production of IL-6, TNF-α, and nitrite in the LPS-stimulated cells. CONCLUSIONS This study showed that the extract of Ej leaves could be used to inhibit the activation of a wide variety of the inflammation-related genes and the inflammatory mediators. It is suggested that the extract of Ej leaves may be applied to alleviate the inflammation of periodontal diseases.

Platycodon grandiflorum A. De Candolle Ethanolic Extract Inhibits Adipogenic Regulators in 3T3-L1 Cells and Induces Mitochondrial Biogenesis in Primary Brown Preadipocytes

This study was designed to evaluate the effects of Platycodon grandiflorum A. DC. ethanolic extract (PG) on obesity in brown/white preadipocytes. The effect of PG on the differentiation and mitochondrial biogenesis of brown adipocytes is still not examined. An in vivo study showed that PG induced weight loss in mice with high-fat-diet-induced obesity. PG successfully suppressed the differentiation of 3T3-L1 cells by down-regulating cellular induction of the peroxisome proliferators activated receptor γ (PPARγ), CCAAT enhancer binding protein α (C/EBPα), lipin-1, and adiponectin but increasing expression of silent mating type information regulation 2 homologue 1 (SIRT1) and the phosphorylation of AMP-activated protein kinase α (AMPKα). The effect of PG on the adipogenic factors was compared with that of its bioactive compound platycodin D. In addition, PG increased expressions of mitochondria-related genes, including uncoupling protein 1 (UCP1), peroxisome proliferator activated receptor-coactivator 1 α (PGC1α), PR domain containing 16 (PRDM16), SIRT3, nuclear respiratory factor (NRF), and cytochrome C (CytC) in primary brown adipocytes. These results indicate that PG stimulates the differentiation of brown adipocytes through modulation of mitochondria-related genes and could offer clinical benefits as a supplement to treat obesity.

Arctigenin Inhibits Lung Metastasis of Colorectal Cancer by Regulating Cell Viability and Metastatic Phenotypes

Arctigenin (ARC) has been shown to have an anti-cancer effect in various cell types and tissues. However, there have been no studies concerning metastatic colorectal cancer (CRC). In this study, we investigated the anti-metastatic properties of ARC on colorectal metastasis and present a potential candidate drug. ARC induced cell cycle arrest and apoptosis in CT26 cells through the intrinsic apoptotic pathway via MAPKs signaling. In several metastatic phenotypes, ARC controlled epithelial-mesenchymal transition (EMT) through increasing the expression of epithelial marker E-cadherin and decreasing the expressions of mesenchymal markers; N-cadherin, vimentin, β-catenin, and Snail. Moreover, ARC inhibited migration and invasion through reducing of matrix metalloproteinase-2 (MMP-2) and MMP-9 expressions. In an experimental metastasis model, ARC significantly inhibited lung metastasis of CT26 cells. Taken together, our study demonstrates the inhibitory effects of ARC on colorectal metastasis.