• Journal of Life Science
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• v.26 no.5
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• pp.584-591
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• 2016

Saururus chinensis is a perennial plants, its flavonoid compound is known to exhibit anti-oxidative activity. This study was aimed to investigate the effect of Water Extract and Solvent Fractions of Saururus chinensis on antioxidant, anti-inflammatory and anti-invasive of Phorbol 12-myristate 13-acetate (PMA)-induced matrix metalloproteinase (MMP-2) and MMP-9 activities. Plant samples were fractionated into hexane, CHCl₃, ethyl acetate, butanol, and water fractions, and each of these was assayed individually. The water fraction showed the highest extraction yield at 9.25%(w/w). Anti-oxidative activity was analyzed by DPPH assay. Cell viability was detected by the MTS assay. Anti-inflammatory activity was assayed by the nitric oxide (NO) production in mouse macrophage Raw 264.7 cells. The activity and mRNA expression of MMP-2 and MMP-9 in human oral squamous carcinoma YD-10B cells were examined by zymography and RT-PCR. As results, MMP-2/-9 activation was increased in PMA induced YD-10B cells. In PMA-treated YD-10B cells, the increased mRNA expression and protein activation of MMP-2/-9 were significantly inhibited in the ethyl acetate fraction. The ethyl acetate fraction showed the highest anti-oxidative activity at 73.38%. The ethyl acetate fraction at non-cytotoxic concentrations significantly exhibited the anti-inflammatory activity of Raw 264.7 cells in dose-dependent manner. In conclusion, these findings demonstrate that the ethyl acetate fraction obtained from a chinensis water extract potentiates a promising therapeutic anti-invasive agent and, therefore, as an anti-cancer drug for cancer prevention and therapy in oral cancer.

• Journal of Life Science
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• v.31 no.12
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• pp.1100-1109
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• 2021

The halophyte Artemisia scoparia is an edible medicinal plant, with insecticidal, anti-inflammatory, anticholesterol, antipyretic, and antibacterial effects. The aim of this study was to assess the inhibitory effect of crude extract and solvent-partitioned fractions obtained from A. scoparia on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated human fibrosarcoma HT-1080 cells using four different activity tests: gelatin zymography, MMP enzyme-linked immunosorbent assay (ELISA), wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot assay. A. scoparia samples were extracted twice with methylene chloride (MC) and twice with methanol (MeOH). After the MC and MeOH crude extracts were combined, the combined crude extracts showed a significant inhibitory effect against MMP-2 and MMP-9 enzymes. They were then fractionated into n-hexane, 85% (v/v) aqueous methanol (85% (v/v) aq.MeOH), n-butanol, and water according to solvent polarity. Among the four solvent-partitioned fractions, n-hexane and 85% (v/v) aq. MeOH fractions significantly inhibited MMP-2 and MMP-9 activity and cell mobility. In addition, the n-hexane and 85% (v/v) aq.MeOH fractions effectively inhibited MMP-2 and -9 activity in the gelatin zymography and MMP ELISA assay. In the wound healing assay, RT-PCR, and Western blot assay, all solvent-partitioned fractions, except the H₂O fraction, significantly suppressed cell migration, as well as the expression levels of MMP-2 and -9 mRNA and proteins.

• BMB Reports
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• v.28 no.6
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• pp.527-532
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• 1995

Human taurine transporter has 12 transmembrane domains and its molecular weight is 69.6 kDa. The long cytoplasmic carboxy and amino termini might function as regulatory attachment sites for other proteins. Six potential protein kinase C phosphorylation sites have been reported in human taurine transporter. In this report, we studied the effects of phorbol 12-myristate 13-acetate (PMA) and glucocorticoid hormone on taurine transportation in the RAW 264.7, mouse macrophage cell line. When the cells were incubated with $[^{3}H]taurine$ in the presence or absence of $Na^+$ ion for 40 min at $37^{\circ}C$, the [$[^{3}H]taurine$ uptake rate was 780-times higher in the $Na^{+}-containing$ buffer than in the $Na^{+}-deficient$ buffer, indicating that this cell line expresses taurine transporter protein on the cell surface. THP1, a human promonocyte cell line, also showed a similar property. The $[^{3}H]taurine$ uptake rate was not influenced by the inflammatory inducing cytokines such as interleukin-1, gamma-interferon or interleukin-1+gamma-interferon, but was decreased by the PMA in the RAW 264.7 cell line. This suggests that activation of protein kinase C inhibits taurine transporter activity directly or indirectly. The inhibition of $[^{3}H]taurine$ uptake by PMA was time-dependent. Maximal inhibition occurred in one hr stimulation with PMA Increasing the treatment time beyond one h reduced the $[^{3}H]taurine$ uptake inhibition due to the depletion or inactivation of protein kinase C. The cell line also showed concentration-dependent $[^{3}H]taurine$ uptake under PMA stimulation. The phorbol-ester caused 23% inhibition at the concentration of 1 ${\mu}m$ PMA. The inhibition was significant even at a concentration as low as 10 nM PMA The reduced $[^{3}H]taurine$ uptake could be recovered by treatment with glucocorticosteroid hormone. Dexamethasone led to recover of the reduced taurine uptake induced by phorbol-ester, recovering maximally after one hr. This may suggest that macrophage cells require higher taurine concentration in a stressed state, for the secretion of glucocorticoid hormone is increased by hypothalamo-pituitary-adrenocortical (HPA) axis activation in the blood stream.

• The Journal of the Korea Contents Association
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• v.20 no.6
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• pp.124-130
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• 2020

The present study investigated the anti-proliferate and anti-invasive of Phorbol 12-myristate 13-acetate (PMA)-induced matrix metalloproteinase (MMP-2) and MMP-9 activities of combined treatment with cisplatin and ethyl acetate fractions of Paeonia japonica. Cell Proliferation was detected by the MTS assay and the activity and mRNA expression of MMP-2/-9 were examined by zymography and RT-PCR. As results, cisplatin or p. japonica treatment of YD-10B cells resulted in a dose-dependent inhibition of cell growth. Also, the viability of YD-10B cells treated with combination of 200 μM cisplatin and 50 ㎍/ml p. japonica was inhibited to 50% in compared with the cisplatin alone. In PMA-treated YD-10B cells, co-treatment of 200 μM cisplatin with 50 ㎍/ml p. japonica significantly inhibited mRNA expression and protein activation of MMP-2/-9. Therefore, This study suggest that the combination treatment of cisplatin and p. japonica potentiates a promising anti-invasive agent and has more potential anti-cancer drug for oral cancer therapy than cisplatin alone.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.3
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• pp.887-892
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• 2004

Cheonggeumganghwa-tang(CGT) has been used for the purpose of prevention and treatment of bronchial asthma and allergic asthma in Korea. To investigate the biological effect of CGT, the author examined cytotoxicity and inflammatory cytokines secretion with human mast cell line, HMC-1. HMC-1 was stimulated with phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. CGT by itself had no effect on viability of HMC-1. The effects of CGT on the secretion of tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6 from HMC-1 were evaluated with enzyme-linked immunosorbent assay (ELISA). CGT (1 ㎎/㎖) inhibited PMA plus A23187 -induced TNF-α and IL-6 secretion, by 93.86 ± 2.05%, 68.69 ± 2.86%, respectively. CGT also inhibited the NF-κB (p50) expression. Taken together, these results suggest that CGT inhibit the production of inflammatory cytokines in HMC-1 cells through blockade of NF-κB activation.

• BMB Reports
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• v.31 no.4
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• pp.350-354
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• 1998

To understand the role of protein kinase C (PKC) in the regulation of chondrogenesis, we examined proteins which are phosphorylated by PKC. Stage 23/24 chick embryo wing mesenchymes were micromass-cultured to induce chondrogenesis and cell extracts were phosphorylated in a condition that activates PKC. Several proteins including 63 and 66 kDa proteins were phosphorylated. The 66 kDa protein was phosphorylated only in the presence of phorbol 12-myristate 13-acetate (PMA) and phosphatidylserine CPS), and the phosphorylation was almost completely diminished by bisindolylmaleimide, a PKC inhibitor. In addition, partially purified PKC increased the phosphorylation of the 66 kDa protein. Treatment of cultures with lysophosphatidylcholine (LPC) promoted chondrogenesis and phosphorylation of 66 kDa protein, while PMA and thymeleatoxin inhibited both of the two events. Our results suggest that the 66 kDa protein is a putative substrate of PKC, and phosphorylation of the 66 kDa protein, probably by $PKC\alpha$ is required for chondrogenesis.

• Advances in Traditional Medicine
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• v.5 no.1
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• pp.69-74
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• 2005

'SecSec' has been used for the purpose of prevention and treatment of throat diseases such as sore throat, cough, bronchial asthma and allergic asthma in Korea. However, its effect in experimental models remains unknown. To investigate the biological effect of SecSec, we examined cytotoxicity and secretion of inflammatory cytokines on human leukemic mast cell line, HMC-1, stimulated with phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. SecSec by itself had no cytotoxicity on HMC-1. When SecSec (1 mg/ml) was added, the secretion of tumor necrosis factor-alpha $(TNF-{\alpha})$, interleukin (IL)-6, and granulocyte macrophage-colony stimulating factor (GM-CSF) was significantly inhibited about 47.20%, 25.55%, and 46.43%, respectively on PMA plus A23187-stimulated HMC-1 cells. But SecSec did not inhibit IL-8 secretion. These findings may help understanding the mechanism of action of this medicine leading to control activated mast cells on allergic inflammatory condition like asthma.

• Natural Product Sciences
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• v.20 no.3
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• pp.196-201
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• 2014

Pyunkang-hwan (Pyunkang-tang) extract (PGT) is a traditional folk medicine for controlling diverse pulmonary diseases including bronchitis, tonsiltis and pneumonitis. We investigated whether PGT significantly affects secretion, production and gene expression of airway mucin using in vivo and in vitro experimental models reflecting the hypersecretion and/or hyperproduction of mucus observed in inflammatory pulmonary diseases. For in vivo experiment, effect of PGT was checked on hypersecretion of pulmonary mucin in sulfur dioxide-induced bronchitis in rats. For in vitro experiment, confluent NCI-H292 cells were pretreated with PGT for 30 min and then stimulated with EGF (epidermal growth factor), PMA (phorbol 12-myristate 13-acetate) or TNF-${\alpha}$ (tumor necrosis factor-${\alpha}$) for 24 h. The MUC5AC mucin gene expression and mucin protein production were measured by RT-PCR and ELISA. The results were as follows: (1) PGT inhibited the expression of MUC5AC mucin gene induced by EGF, PMA or TNF-${\alpha}$ from NCI-H292 cells, respectively; (2) PGT also inhibited the production of MUC5AC mucin protein induced by the same inducers from NCI-H292 cells, respectively; (3) PGT inhibited secretion of mucin in sulfur dioxide-induced bronchitis rat model. This result suggests that PGT can regulate secretion, production and gene expression of airway mucin.

• Natural Product Sciences
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• v.16 no.3
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• pp.185-191
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• 2010

Mast cell-mediated allergic disease is involved in many diseases such as anaphylaxis, asthma and atopic dermatitis. The discovery of drugs for the treatment of allergic disease is an important subject in human health. In the present study, the effect of water extract of Gleditsiae Spina (WGS) (Leguminosae), on compound 48/80-induced systemic allergic reaction, anti-DNP IgE antibody-induced local allergic reaction, and histamine release from human mast cell line (HMC-1) cells were studied. In addition, the effect of WGS on phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187 (A23187)-induced gene expression and secretion of pro-inflammatory cytokines were investigated using HMC-1 cells. WGS was anally administered to mice for high and fast absorption. WGS inhibited compound 48/80-induced systemic allergic reaction. WGS dose-dependently decreased the IgE-mediated passive cutaneous anaphylaxis. WGS reduced histamine release from HMC-1 cells. In addition, WGS decreased the gene expression and secretion of pro-inflammatory cytokines in PMA plus A23187-stimulated HMC-1 cells. These findings provide evidence that WGS could be a candidate as an antiallergic agent.

• Food Science of Animal Resources
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• v.25 no.2
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• pp.244-249
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• 2005

To investigate the effect of Holstein colostrum peptide fraction on proliferation of immune cell, polypeptide fractions were separated from acid whey into 3 fractions depending on molecular weight by ultrafiltration: Fraction I, which contains the polypeptide larger than 10,000 Da, Fraction n, which contains the polypeptide ranging from 1,000 Da to 10,000 Da and Fraction III, which contains the polypeptide smaller than 1,000 Da. EL-4 cell (murine T lymphoma cell) was used to evaluate immune enhancing effect of each fraction from Holstein colostrum. Fraction n showed the highest proliferative effect of the colostrum whey fractions on EL-4 cell at 1mg/mL compared with whole whey and other fractions and this proliferative activity was shown in dose dependent manner. Fraction n showed the highest proliferative effect on PMA (Phorbol 12-myristate 13-acetate) stimulated EL-4 cell. Heated Fraction n showed similar effect to native one on proliferation of both EL-4 cell and PMA stimulated EL-4 cell.