• The Journal of Korean Obstetrics and Gynecology
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• v.36 no.3
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• pp.1-24
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• 2023

Objectives: Boeum-jeon (BEJ) is a herbal formula composed 8 Korean medicinal herbs and is traditionally used to treat inflammatory diseases. In this study, the authors tried to confirm the antioxidant efficacy of BEJ and its anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Methods: In this experiment, results of BEJ on the following two were measured as follows: (1) Antioxidant effects was measured by DPPH Radical scavenging Activity, ABTS Radical scavenging Activity. (2) Anti-Inflammatory effects were evaluated by the production amount of ROS, NO, Cytokine (PGE2, IL-1β, IL-6, TNF-α), COX-2, iNOS, TNF-α, IL-1β, IL-6, HO-1, NQO1 (the previous seven are "mRNA Expression"), COX-2, iNOS, TNF-α, IL-1β, IL-6, HO-1, NQO1, NRF2 (the previous eight are "Protein Expression") ERK, JNK, p38 (the previous three are "Protein Phosphorylation") in LPS-stimulated RAW 264.7 cells. Results: The experimental measurement results are as follows: (1) DPPH Radical scavenging Activity, ABTS Radical scavenging Activity increased in a concentration -dependent manner in the BEJ-treated group. (2) As a result of measuring anti-inflammatory efficacy, the production of ROS, NO, and Cytokine (PGE2, TNF-α, IL-1β, IL-6) in the BEJ-administered group was significantly reduced compared to the control group. (3) Among mRNA Expression levels, COX-2, iNOS, IL-1β, IL-6 and TNF-α was significantly decreased in a concentration-dependent manner than in the control group, and HO-1 and NQO1 were significantly increased in a concentration -dependent manner than in the control group. (4) Among the Protein Expression levels, COX-2, iNOS, IL-1β, IL-6 and TNF-α was significantly decreased in a concentration -dependent manner compared to the control group, and HO-1, NQO1 and NRF2 was significantly increased in a concentration-dependent manner compared to the control group. (5) As a result of Protein Phosphorylation, ERK, p38 and JNK was significantly decreased compared to the control group in a concentration-dependent manner. Conclusions: Boeum-jeon has been experimentally confirmed to have antioxidant and anti-inflammatory effects, and if the evidence for efficacy is reinforced through further studies such as in vivo studies and clinical trials in the future, it can be effectively used to treat various inflammatory diseases such as bladder inflammation and chronic pelvic inflammation.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.3
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• pp.941-945
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• 2015

Semaphoring is a transmembrane receptor which participates in many cytokine-mediated signal pathways that are closely related to the angiogenesis, occurrence and development of carcinoma. The present study was designed to access the effect of mono-antibody (mAb) guided radioimmunotherapy (RIT) on skin carcinoma and investigate the potential mechanisms. Semaphoring mAb was acquired from mice (Balb/c), purified with rProtein A column; purity, concentration and activity were tested with SDS-PAGE and indirect ELISA; specificity and expression on the cutanuem carcinoma line and tissue were tested by Western blotting; morphology change was assessed by microscopy. MTT assay and colony inhibition tests were carried out to test the influence on the proliferation of tumor cells; Western blotting was also carried out for expression of apoptosis-associated (caspase-3, Bax, Bcl-2) and proliferation-related (PI3K, p-Akt, Akt, p-ERK1/2, ERK1/2) proteins and analyse the change in signal pathways (PI3K/Akt and MEK/ERK). The purity of purified semaphorin mAb was 96.5% and the titer is about $1{\times}10^6$. Western blotting showed semaphoring mAb to have specifically binding stripes with semaphoring b1b2 protein, B16F10, and A431 cells at 39KDa, 100KDa and 130KDa, respectively. Positive expression was detected both in cutanuem carcinoma line and tissue and it mostly located in cell membranes. MMT assay revealed dose-relate and time-relate inhibitory effect of semaphorin mAb on A431 and B16F10. Colony inhibition tests also showed dose-relate inhibitory effects. Western blotting demonstrated the expression of apoptosis and proliferation-related protein and changes in signal pathway. In conclusion, we demonstrated that semaphorin is highly expressed on the tumor cell-surfaces and RIT with semaphorin mAb has effect in i nhibiting proliferation and accelerating apoptosis of tumor cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.7
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• pp.896-902
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• 2017

In this study, we investigated citrus Kombucha (CK) produced by three different bacteria strains (Gluconacetobacter xylinus, Gluconacetobacter medellinensis, and Gluconobacter oxydans; named as CK-MOX) identified from traditional Kombucha. During fermentation, the pH level of CK-MOX was gradually reduced, and total acidity slightly increased. Antioxidant activity, measured by DPPH, ABTS, and oxygen radical absorbance capacity assays, markedly increased after fermentation. Moreover, fermented CK-MOX (Day15) exhibited anti-proliferative and anti-migratory activities against EJ human bladder carcinoma cells. Western immunoblot assays showed that treatment with CK-MOX significantly up-regulated phospho-extracellular signaling kinase (ERK) levels. To distinguish whether or not up-regulation of phospho-ERK is the cause or effect, we investigated the viability of EJ cells in the presence of U0126, a mitogen activated protein kinase/ERK kinase 1/2 inhibitor. Pre-treatment with U0126 rescued cells from CK-MOX-induced cell death, which indicates phospho-ERK may be a key regulator in the mechanism of CK-MOX-induced apoptosis of EJ bladder cancer cells. In conclusion, CK-MOX, fermented by a defined composition of bacterial starters, shows antioxidant capacity and anti-cancer activity against EJ bladder cancer cells.

• The Journal of Internal Korean Medicine
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• v.32 no.2
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• pp.217-231
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• 2011

Objectives : This study aimed to evaluate the effects of Saengmaekcheongpye-eum (SCE) on a LPS-induced COPD (chronic obstructive pulmonary disease) model. Materials and Methods : The extract of SCE was treated to A549 cells and and LPS-induced COPD mouse model. Then, various parameters such as cell-based cyto-protective activity and histopathological finding were analyzed. Results : SCE showed a protective effect on LPS-induced cytotoxicity in A549 cells. This effect was correlated with analysis for caspase 3 levels, elastin contents, protein levels of cyclin B1, Cdc2, and phospho-Erk1/2, and gene expression of TNF-${\alpha}$ and IL-$1{\beta}$ in A549 cells. SCE treatment also revealed a protective effect on LPS-induced lung injury in COPD mouse model. This effect was evidenced via histopathological findings including immunofluorescence stains against elastin and caspase 3, and protein levels of cyclin B1, Cdc2, and Erk1/2 in lung tissue. Conclusions : These data suggest that SCE has pharmaceutical properties on lung injury. This study thus provides scientific evidence for the efficacy of SCE for clinical application to patients with COPD.

• Archives of Pharmacal Research
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• v.29 no.11
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• pp.1024-1031
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• 2006

The role of mitogen-activated protein kinase (MAPK) in the decreased contractile response to phorbol ester in aortic smooth muscle strips from deoxycorticosterone acetate (DOCA)-salt hypertensive rats was examined. Norepinephrine (NE) evoked greater contractility in aortic strips from DOCA rats than in those of sham-operated rats. 12-Deoxyphorbol 13-isobutyrate (DPB) induced contraction in $Ca^{2+}-free$ medium, which was diminished in strips from DOCA rats compared to sham-operated rats. Vasoconstrictions induced by these stimulants were inhibited by SB203580 and PD098059, inhibitors of p38 MAPK and extracellular signal-regulated kinase (ERK) 1/2, respectively, in both strips. The phosphorylation of p38 MAPK and ERK1/2 induced by NE was greater in strips from DOCA rats compared to those from sham-operated rats, and this phosphorylation was inhibited by the kinase inhibitors. DPB increased the phosphorylation of p38 MAPK and ERK1/2 in strips from both animals, and the increment of p38 MAPK phosphorylation by the stimulant was diminished in strips from DOCA rats compared to sham-operated rats. These findings suggest that the $Ca^{2+}-independent$ contraction evoked by DPB results from the activation of MAPKs in rat aortic smooth muscle and that the attenuated contractility by DPB in DOCA rat appears to be associated with diminished p38 MAPK activity.

• Toxicological Research
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• v.21 no.4
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• pp.347-353
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• 2005

Eukaryotic initiation factor 4E (elF4E) is a key element for cap-dependent protein translation controlled by affinity between elF4E and 4E-binding protein 1 (4E-BP1). Rapamycin can also affect protein translation by regulating 4E-BP1 phosphorylation. Tobacco-specific nitrosamine, 4(N-methyl-N-nitrosamino )-1-(3-pyridyl)-1-butanone (NNK) is a strong lung carcinogen, but its precise lung cancer induction mechanism remains unknown. Relative roles of cap-dependent and -independent protein translation in terms of NNK-induced lung carcinogenesis were elucidated using normal human bronchial epithelial cells. NNK concentrations applied in this study did not decrease cell viability. Addition of NNK restored rapamycin-induced decrease of protein synthesis and rapamycin-induced phosphorylation of 4E-BP1, and increased expression levels of mTOR, ERK1/2, p70S6K, and Raf-1 in a concentration-dependent manner. NNK also caused perturbation of normal cell cycle progression. Taken together, NNK might cause toxicity through the combination of restoration of 4E-BP1 phosphorylation and increase of elF4E as well as mTOR protein expression, interruption of Raf1/ERK as well as the cyclin G-associated p53 network. Our data could be applied towards elucidation of the molecular basis for lung cancer treatment.

• Nutrition Research and Practice
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• v.17 no.5
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• pp.844-854
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• 2023

BACKGROUND/OBJECTIVES: Fucoidan, a polysaccharide content in brown algae, has been reported to inhibit the growth of cancer cells. The present study aimed to investigate the suppression effects of fucoidan on A549 non-small cell lung cancer cells migration. MATERIALS/METHODS: The anti-migratory activity of fucoidan in A549 cells was examined by wound healing assay and phalloidin-rhodamine staining in response to fucoidan (0-100 ㎍/mL) treatment for 48 h. Western blot analysis was performed to clarify the protein expressions relevant to migratory activity. RESULTS: Fucoidan (25-100 ㎍/mL) significantly suppressed A549 cells migration together with reduced the intensity of phalloidin-rhodamine which detect filopodia and lamellipodia protrusions at 48 h of treatment. The protein expression indicated that fucoidan significantly suppressed the phosphorylation of focal adhesion kinase (FAK), Src, and extracellular signal-related kinase (ERK). In addition, the phosphorylation of p38 in A549 cells was found to be increased. CONCLUSIONS: Our data conclude that fucoidan exhibits anti-migratory activities against lung cancer A549 cells mediated by inhibiting ERK1/2 and FAK-Src pathway.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.76.2-76.2
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• 2003

Although sphingosine-1-phosphate (S1P) is a well-known mitogen, our results show that S1P potently inhibits keratinocyte proliferation, and that this leads the inhibition of DNA synthesis. Interestingly, the prolonged activation of extracellular signal-regulated protein kinase (ERK) and the transient inactivation of Akt/protein kinase B (PKB) were also oberved in concert with the inhibition of keratinocyte proliferation by S1P. To further verify the anti-proliferative action of S1P, we examined changes in cell cycle related proteins. (omitted)

• Korean Journal of Veterinary Research
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• v.44 no.1
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• pp.7-13
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• 2004

Severe hypertension (>180 mmHg) develops in spontaneously hypertensive rats (SHR) after 12 wk-old; however, it is not clear whether what kinds of molecular mechanism leads to altered cardiac performance following developmental stages in SHR. Also, although the effect of calcineurin (Cn) to promote cardiomyocyte hypertrophy in vivo and in vitro is established, its overall necessity as a hypertrophic mediator is currently an area of ongoing debate. Thus, we have examined i) body weight and blood pressure, ii) differences of expression and distribution of signaling molecules such as Cn, protein kinase B/Akt (PKB/Akt), and extracellular signal-regulated kinase (ERK) between SHR and their age-matched control Wistar-Kyoto (WKY) rats following developmental stages. In 16 wk-old SHR compared with WKY, 2-dimentional echocardiography showed cardiac enlargement and hypertrophy of left ventricle, significantly. Taken together, we suggest that Cn is associated with hereditary cardiac hypertrophy, the process being related to the molecular signaling mechanisms involving PKB/Akt and ERK.

• Journal of Physiology & Pathology in Korean Medicine
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• v.34 no.1
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• pp.24-29
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• 2020

In this study, we investigated the inhibition effects of mixtures of Atractylodes macrocephala (AM) and Amomum villosum (AV) water extracts on adipocyte differentiation. Treatment with mixtures of AM and AV extracts in a ratio of 3:1 for 24 and 48 hours did not show any cytotoxicity in OP9 cells. Mixtures of AM(3) and AV(1) extracts inhibited adipocyte differentiation, expression of peroxisome proliferator-activated receptor gamma (PPARγ) and CCAT/enhancer-binding protein alpha (C/EBPα), the major transcription factors of differentiation. It also inhibited the expression of lipoprotein lipase (LPL), adipocyte protein 2 (aP2), which are PPARγ-target genes in adipocyte. We also checked the inhibition effects on cell proliferation during the early stage of differentiation by treatment with mixtures of AM(3) and AV(1) extracts. It markedly inhibited adipocyte proliferation after 48 hours, and also the phosphorylation of ERK1/2 and Akt after 10 min or 3 hour. These results identify a possible mechanism of action of mixtures of AM(3) and AV(1) extracts, suggesting that the mixtures of AM(3) and AV(1) extracts-induced inhibition of ERK and Akt phosphorylation suppresses adipogenesis by inhibiting other signaling cascades that include PPARγ and C/EBPα during the process of OP9 adipocyte differentiation.