• The Journal of Internal Korean Medicine
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• v.29 no.2
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• pp.469-489
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• 2008

Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.

• Journal of Nutrition and Health
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• v.53 no.2
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• pp.111-120
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• 2020

Purpose: Corosolic acid (CA), also known as 2α-hydroxyursolic acid, is present in numerous plants, and is reported to exhibit anti-cancer and anti-proliferative activities in various cancer cells such as osteosarcoma, hepatocellular carcinoma, lung adenocarcinoma, and colon cancer. However, the anti-cancer activity of CA on human breast cancer cells and the underlying mechanisms remain to be elucidated. The present study aimed to investigate the anticancer effects of CA in the human breast cancer cell line, MDA-MB-231. Methods: Cell viability, reactive oxygen species (ROS) production, apoptosis marker protein expression, migration, invasion rate, and vascular endothelial growth factor (VEGF) levels were assessed by treating MDA-MB-231 cells to increasing concentrations of CA. Results: The results showed that CA significantly inhibited the cell proliferation of MDA-MB-231 cells in a dose-dependent manner. To assess the effect of CA on apoptosis, nuclei of MDA-MB-231 cells were stained with DAPI solution. Chromatin condensation, which indicates apoptosis, was observed to increase dose-dependently. In addition, western-blot analysis revealed elevated levels of the apoptosis marker proteins (Bax and cleaved caspase 3) subsequent to MDA-MB-231 exposure to CA. ROS production was also increased in the CA-induced apoptosis in MDA-MB-231 treated cells. Interestingly, CA exposure resulted in significantly decreased migration and invasion rates in the MDA-MB-231 cells. Data further revealed that exposure to CA markedly decreased the VEGF concentration, thereby contributing to a reduction in angiogenesis. Conclusion: Our results determined that exposure to CA induces anti-proliferation, apoptosis, and ROS production, and suppresses cell migration and invasion rate in MDA-MB-231 cells. Taken together, these results indicate the potential of CA to be applied as an effective chemotherapeutic agent for treating breast cancer.

• Herbal Formula Science
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• v.25 no.4
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• pp.509-526
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• 2017

Background and objectives : Soeumin Bojungykgi-tang (seBYTE) has been used to supplement qi in Korean medicine. It has been demonstrated to possess various biological functions such as anti-cancer, anti-aging and anti-inflammatory effects. The present study evaluated the protective roles of seBYTE in hepatotoxic in vitro and in vivo model. Methods : To investigate cytoprotective effect of seBYTE, HepG2 cells were pretreated with seBYTE and then subsequently exposed to $10{\mu}m$ AA for 12 h, followed by $5{\mu}m$ iron. Cell viability was examined by MTT assay, and expression of apoptosis-related proteins was evaluated by immunoblot analysis. For responsible molecular mechanisms, ROS production, GSH contents, and mitochondrial membrane potential were measured. In addition, hepatoprotective effect of seBYTE in vivo was assessed in $CCl_4$-induced animal model. Results : seBYTE prevented AA + iron-induced cytotoxicity in concentration dependent manner. In addition, ROS production, GSH depletion, and mitochondrial dysfunction induced by AA + iron were significantly reduced by seBYTE pretreatment. Furthermore, seBYTE recovered expression of the pro-apoptotic proteins such as PARP and pro-caspase-3. In animal experiment, plasma ALT and AST levels were significantly elevated in $CCl_4$ treatment, but seBYTE significantly decreased the ALT and AST levels. Moreover, seBYTE alleviated the numbers of histological activity index, percentages of degenerative regions, degenerated hepatocytes, infiltrated inflammatory cells, nitrotyrosine- and 4-hydroxynonenal-positive cells in liver. Conclusions : These results showed that hepatoprotective effect of seBYTE against on $CCl_4$-induced hepatic damages is partly due to antioxidative and anti-apoptotic process.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.978-984
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• 2004

In the course of screening for a novel inhibitor of CDC2, HY558-1 was isolated from a culture broth of Penicillium minioluteum F558. Moreover, it was found that HY558-1 had an effect on both the cell cycle regulation and apoptosis of human cervical adenocarcinoma HeLa cells. A flow cytometric analysis of HeLa cells revealed appreciable cell cycle arrest at the G1 and G2/M phases following treatment with HY558-1. Furthermore, DNA fragmentation due to apoptosis was observed in HeLa cells treated with HY558-1. To obtain further information on the cell cycle arrest and apoptotic induction induced by HY558-1, the expression of certain cell cycle and apoptosis-associated proteins was examined using a Western blot analysis. The results revealed that HY558-1 inhibited the phosphorylation of pRb and decreased the expression levels of CDK2, CDC2, and cyclin A in the cell cycle progression. It was also shown that the level of $p21^{WAF1/CIP1}$ was increased in HeLa cells treated with 0.52 mM of HY558-1. Accordingly, HY558-1 was found to inhibit the proliferation of HeLa cells through the induction of G1 phase arrest by inhibiting pRb phosphorylation via an upregulation of $p21^{WAF1/CIP1}$, and G2/M phase arrest by directly inhibiting CDC2 and cyclin A. Moreover, HeLa cells treated with 0.52 mM of HY558-1 exhibited apoptotic induction associated with the cleavage of Bid and release of cytochrome c from mitochondria into the cytosol. Subsequent investigation of the activation of caspase-3 and cleavage of poly (ADP-ribose) polymerase (PARP) suggested that the mitochondrial pathway was primarily involved in the HY558-1-induced apoptosis in HeLa cells.

• CELLMED
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• v.3 no.2
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• pp.18.1-18.7
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• 2013

Inflammation in rheumatoid arthritis is characterized by immune cell infiltration and cytokine secretion. In particular, mast cells and their cytokines play an important role in the pathogenesis of rheumatoid arthritis. Korean medicine, BaekJeol-Tang (BT) was designed by traditional Korean medicine theory. We already reported therapeutic effect of BT in rheumatoid arthritis. Here, we report the specific underlying mechanism of BT in activated human mast cells, HMC-1 cells. In addition, we report for the first time that BT significantly inhibited the production and mRNA expression of proinflammatory cytokines including thymic stromal lymphopoietin, interleukin (IL)-$1{\beta}$, IL-6, IL-8, and tumor necrosis factor-${\alpha}$ in activated HMC-1 cells. BT also decreased the activation of mitogen-activated protein kinases, nuclear factor-${\kappa}B$, and caspapase-1. Taken together, these results indicate that BT has potential as a regulator of inflammatory reactions for the treatment of arthritis such as osteoarthritis and rheumatoid arthritis.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.7
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• pp.3035-3042
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• 2015

Background: Isorhamnetin (Iso), a novel and essential monomer derived from total flavones of Hippophae rhamnoides that has long been used as a traditional Chinese medicine for angina pectoris and acute myocardial infarction, has also shown a spectrum of antitumor activity. However, little is known about the mechanisms of action Iso on cancer cells. Objectives: To investigate the effects of Iso on A549 lung cancer cells and underlying mechanisms. Materials and Methods: A549 cells were treated with $10{\sim}320{\mu}g/ml$ Iso. Their morphological and cellular characteristics were assessed by light and electronic microscopy. Growth inhibition was analyzed by MTT, clonogenic and growth curve assays. Apoptotic characteristics of cells were determined by flow cytometry (FCM), DNA fragmentation, single cell gel electrophoresis (comet) assay, immunocytochemistry and terminal deoxynucleotidyl transferase nick end labeling (TUNEL). Tumor models were setup by transplanting Lewis lung carcinoma cells into C57BL/6 mice, and the weights and sizes of tumors were measured. Results: Iso markedly inhibited the growth of A549 cells with induction of apoptotic changes. Iso at $20{\mu}g/ml$, could induce A549 cell apoptosis, up-regulate the expression of apoptosis genes Bax, Caspase-3 and P53, and down-regulate the expression of Bcl-2, cyclinD1 and PCNA protein. The tumors in tumor-bearing mice treated with Iso were significantly smaller than in the control group. The results of apoptosis-related genes, PCNA, cyclinD1 and other protein expression levels of transplanted Lewis cells were the same as those of A549 cells in vitro. Conclusions: Iso, a natural single compound isolated from total flavones, has antiproliferative activity against lung cancer in vitro and in vivo. Its mechanisms of action may involve apoptosis of cells induced by down-regulation of oncogenes and up-regulation of apoptotic genes.

• BMB Reports
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• v.45 no.12
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• pp.742-747
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• 2012

Granulocyte colony-stimulating factor (G-CSF) is used for heart failure therapy and promotes myocardial regeneration by inducing mobilization of bone marrow stem cells to the injured heart after myocardial infarction; however, this treatment has one weakness in that its biological effect is transient. In our previous report, we generated 5 mutants harboring N-linked glycosylation to improve its antiapoptotic activities. Among them, one mutant (Phe140Asn) had higher cell viability than wild-type hG-CSF in rat cardiomyocytes, even after treatment with an apoptotic agent ($H_2O_2$). Cells treated with this mutant significantly upregulated the antiapoptotic proteins, and experienced reductions in caspase 3 activity and PARP cleavage. Moreover, the total number of apoptotic cells was dramatically lower in cultures treated with mutant hG-CSF. Taken together, these results suggest that the addition of an N-linked glycosylation was successful in improving the antiapoptotic activity of hG-CSF, and that this mutated product will be a feasible therapy for patients who have experienced heart failure.

• Korean Journal of Veterinary Research
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• v.55 no.2
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• pp.133-139
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• 2015

The increasing uses of zinc oxide nanoparticles (nZnO) in industrial and personal care products raise possible danger of using nZnO in human. To determine whether ZnO induces size-dependent anomalies during embryonic organogenesis, mouse embryos on embryonic day 8.5 were cultured for 2 days under 50, 100, and $150{\mu}g$ of nZnO (< 100 nm) or micro-sized ZnO (mZnO; $80{\pm}25{\mu}m$), after which the morphological changes, cumulative quantity of Zn particles, and expressions of antioxidant and apoptotic genes were investigated. Although embryos exposed to $50{\mu}g$ of ZnO exhibited no defects on organogenesis, embryos exposed to over $100{\mu}g$ of ZnO showed increasing anomalies. Embryos treated with $150{\mu}g$ of nZnO revealed significant changes in Zn absorption level and morphological parameters including yolk sac diameter, head length, flexion, hindbrain, forebrain, branchial bars, maxillary process, mandibular process, forelimb, and total score compared to the same dose of mZnO-treated embryos. Furthermore, CuZn-superoxide dismutase, cytoplasmic glutathione peroxidase (GPx) and phospholipid hydroperoxidase GPx mRNA levels were significantly decreased, but caspase-3 mRNA level was greatly increased in nZnO-treated embryos as compared to normal control embryos. These findings indicate that nZnO has severer teratogenic effects than mZnO in developing embryos.

• Herbal Formula Science
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• v.28 no.1
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• pp.1-13
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• 2020

Objectives : Present study investigated hepatoprotective effects of four types of Taraxaci Herba water extract (TL-F, Taraxaci Herba leaf originated from foreign country; TR-F, Taraxaci Herba root originated from foreign country; TL-K, Taraxaci Herba leaf originated from Korea; TR-K, Taraxaci Herba root originated from Korea) on carbon tetrachloride (CCl₄)-induced acute liver injury. Methods : Mice were administered orally with 200 mg/kg of TL-F, TR-F, TL-K, or TR-K for seven days, and intraperitoneally injected with 0.5 mL/kg of CCl₄ 1 h after last Taraxaci Herba treatment. Silymarin (100 mg/kg) was used as a positive drug. Body weight gain, relative liver weight, serum biochemistry, histopathological and immunohistochemical analyses, and hepatic antioxidant capacity were determined to investigate the hepatoprotective effect of four extracts. Results : Administration of four types of Taraxaci Herba extract increased body weight gain, and decreased relative liver weight in CCl₄-injected mice. As compared to CCl₄ group, TL-F and TR-F significantly decreased serum aspartate aminotransferase activity, while four extracts reduced CCl₄-induced alanine aminotransferase activity. In addition, TL-F and TR-F significantly decreased the numbers of degenerated hepatocytes, infiltrated inflammatory cells, cleaved caspase-3 positive cells, and cleaved PARP positive cells in hepatic tissues. Moreover, TL-F, TR-F, and TR-K administration reduced the lipid peroxidation and nitrotyrosine, and increased glutathione, superoxide dismutase, and catalase activities in hepatic tissues. There were no statistical differences between TL-F- and silymarin-treated group. Conclusion : Of four extracts tested, present results suggest that TL-F is the most favorable candidate against CCl₄-induced acute liver injury through enhancing antioxidant activity.

• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.147-152
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• 2020

Farrerol is a flavanone isolated from the traditional Chinese herb 'Man-shan-hong' (Rhododendron dauricum L.). Farrerol has been reported to have various bioactivities including anti-oxidant, anti-inflammation, and anti-fungal. However, anti-cancer effect of farrerol has not yet been reported in MCF-7 breast cancer cells. In the present study, we investigated the anti-cancer effect of farrerol on MCF-7 cells. Farrerol decreased viability and induced apoptosis of MCF-7 cells in a dose dependent manner. Ferrerol exhibited a significant anti-proliferation effect with a half-maximal inhibitory concentration (IC₅₀) values of 145.04±1.4 μM in MTT assay, when MCF-7 cells were treated with ferrerol for 48 h. Also, ferrerol induced apoptotic bodies of MCF-7 cells as evaluated by TUNEL assay and Annexin V/PI staining using FACS. By mechanism of action, ferrerol regulated the activation of p38 mitogen-activated protein kinase and altered the expression level of BAX, Bcl-2, and Poly ADP Ribose Polymerase in MCF-7 cells. In summary, our finding demonstrated that ferrerol has anti-cancer effect through regulating the activation and expression of apoptosis-related proteins in MCF-7 cells.