• Korean Journal of Medicinal Crop Science
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• v.14 no.4
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• pp.195-201
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• 2006

The fibrinolytic activities of soluble proteins extracted from young leaves of Camellia japonica L. were studied. Fibrinolvity activity of extract from partitions of C. japonica L. showed 1.6-2.0 times higher than plasmin used as positive control. The fibrinolytic enzyme was confirmed directly from young leaves of C. japonica L. by a fibrin Plate and fibrin zymography. The protein was composed of a single polypeptide and its apparent molecular weight was found to be 45 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for the fibrinolytic activity were pH 5.5 and $30^{\circ}C$, respectively. Also, the fibrinolytic activity was clearly inhibited by PMSF and TLCK, suggesting that it is a member of the trypsin-like serine protease. All these results suggest the protease is a fibrinolytic enzyme belong to a family of trypsin-like serine protease.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.1
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• pp.35-44
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• 2014

This study was performed to investigate the anti-photoaging effects of Persimmon leaf tea(PLT) in hairless mice(SKH-1) exposed to UVB radiation. The animals were divided into non-treated group (normal, N) and UV-radiated groups. UV-radiated groups were divided into only UV-radiated group(control, C) and UV-radiated and PLT treated experimental groups[first extraction treated group(PLT-I), second extraction treated groupe(PLT-II), and third extraction treated group(PLT-III)]. Three PLT treated experimental groups of mice were treated with both oral administration(300mg/Kg B.W./day) and topical application (100 ul of 2% conc./mouse/day) for 4 weeks. Anti-photoaging effects of Persimmon leaf were evaluated by MTT assay, anti oxidative reaction, MMP immunohistochemistry, gelatin zymography assay and RT-PCR observations. Treatment with Persimmon leaf tea(PLT)-I, and -III groups decreased immunohistochemical density of matrix metalloproteinases(MMP)-3 and -9 related to degradation of extracellular matrix in skin. Especially, immunohistochemical density of MMP-2 decreased in PLT-I, -II and -III groups in skin. On the effects of antioxidant function on the treatment with Persimmon leaf tea(PLT), treatment of HaCaT cells with extracts of PLT-I and PLT-II had also significantly reduced intracellular ROS produced by UVB irradiation in a dose dependent manner(PLT-I, p<0.05, p<0.01, p<0.001; PLT-II, p<0.01, p<0.001). Gelatin zymography assay revealed that PLT-II and PLT-III (200 ug/ml) had inhibitory effect on MMP-9 expression in UVB-radiated HaCaT cells. Western blot analysis revealed that PLT-1, -II and -III groups down-regulates the expression of inflammatory associated genes(IL-$1{\beta}$) and PLT-1 and -II groups down-regulates the expression of TNF-${\alpha}$ in a dose dependent manner. Our study suggests that Persimmon leaf tea(PLT) extracts participates in inhibitory effects on the morphological and molecular experiments related to photoaging skin on UVB irradiated hairless mice.

• Journal of Life Science
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• v.21 no.11
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• pp.1501-1510
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• 2011

In recent years novel potential pharmocological candidates have been looked for in animal, seaweed, sponge, fungi and marine bacteria resources. In this study, matrix metalloproteinases (MMPs) that play an important role in metastasis, arthritis, chronic inflammation and wrinkle formation were used as target enzymes to screen therapeutic agents. The inhibitory effects of several marine algae including green algae (5 species), red algae (18 species) and brown algae (4 species) methanolic extracts on MMPs were investigated in human dermal fibroblasts and human fibrosarcoma cell line (HT1080 cells) using gelatin zymography. In human dermal fibroblasts, the inhibition of MMP-2 was observed in Laurencia okamurae, Polysiphonia japonica, Grateloupia lanceolate and Sinkoraena lancifolia of red algae. In contrast, MMP-2 activation was enhanced in Enteromorpha compressa and E. linza of green algae, and Peltaronia bighamiae and Sargassum thunbergii of brown algae. In human fibrosarcoma cells, MMP-9 activation was decreased in the presence of S. thunbergii of brown algae, Polysiphonia japonica in red algae and E. compressa and E. linza of green algae. The interesting finding is that E. compressa and E. linza of green algae, and S. thunbergii of brown algae exhibited a positive effect on MMP-2 in normal cells, but a negative effect on MMP-9 in cancer cell lines. These results suggest that E. compressa and E. linza of green algae, and S. thunbergii of brown algae contain potential therapeutic ingredients for cancer treatment.

• Journal of Korean Society of Forest Science
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• v.18 no.1
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• pp.17-22
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• 1973

Using of zymography for identification of same clone of Pinus densiflora, the two year old needle leaves of 48 ramtes including 8 clones (6 ramets per clone) were collected in the clone bank which has been established on the near campus of Institute of Forest Genetics, Suwon, in 1962. All 8 bands are named from cathod to anode, G, H, K, M, N, Q, S and Y. Only CB-1 clone shows all bands, while KW-3 clone reveals only 5 bands. Other 6 clones were found 7 bands but the occurred frequencies of those bands are variable among those clones. Though the grafting stock are used various individuals grown seed propagation of P. densiflora, moreover, three stocks have been different species and that one has been P. rigida and two individuals have been P. koraiensis, the zymograms of the ramets belonging to the same clone reveals the identified patterns. The results show that the stocks for grafting have not been affected on the isoperoxidase patterns of their scions in P. densiflora. Among 48 ramets of 8 clones, 4 ramets are found the different isoperoxidase patterns from that of the remained rametes within same clone. Thus, it is conluded that zymography is useful for testing genuineness of the grafted clones of P. densiflora.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.224-224
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• 2004

Plasminogen activators (PAs)는 자궁분비액, 난포액, 정장물질 등을 포함한 여러 가지 세포외 분비액(extracellular fluids) 및 plasma에 풍부하게 존재하는 세포외 전구효소인 plasminogen을 plasmin으로 전환시키는 단백질분해효소이다. PAs는 섬유소용해, 배란, 착상 및 수정을 포함한 다양한 생리적인 과정에서 중요한 역할을 수행하는 것으로 알려져 있다. 따라서, 본 연구는 돼지의 체외수정 과정시 정자와 난자에서의 plasminogen activators activity의 변화를 SDS-PAGE와 zymography를 이용하여 검토하였다. (중략)

• YAKHAK HOEJI
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• v.50 no.4
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• pp.228-233
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• 2006

Matrix metalloproteinase-2 is known to be involved in pathological processes such as tumor invasion or rheumatoid arthritis. A soil microorganism producing siderophore under low iron stress $(up\;to\;5\;{\mu}m\;of\;iron)$ was identified as Burkholderia sp. Hydroxamate type siderophore produced by Burkholderia sp. CAS-5 was partially purified. MMP inhibitory activity of siderophore was confirmed by gelatin zymography. The $Zn^{2+}-chelating$ activity of siderophore correlated with the inhibition of MMP-2 activity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.5
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• pp.520-529
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• 2014

The purpose of this study is to investigate the protective effects of Yukmijiwhang-tang(YM) water extracts against the UVB irradiation on the human keratinocyte HaCaT cells. We observed the effects of YM on the oxidative stress, gene expression of pro-inflammatory cytokine such as TNF-${\alpha}$ and IL-$1{\beta}$, and matrix metalloproteinase-9 in UVB-irradiated HaCaT cells. On the effects of oxidative stress and antioxidant function on the treatment with YM, The activity of xanthine oxidase(XO) was significantly decreased by treatment of YM in all the concentrations(p<0.01). The activity of superoxide dismutase(SOD) and catalase(CAT) was significantly increased by treatment of YM in a dose dependent manner(p<0.05 and p<0.01). DPPH radical was erased by treatment of YM under dose of $500{\mu}g/m{\ell}$ concentration. Treatment of HaCaT cells with YM had also significantly reduced intracellular ROS produced by UVB irradiation in a dose dependent manner(p<0.05, p<0.01, p<0.001). Gelatin zymography assay showed that YM downregulated the MMP-9 activity in UVB-irradiated HaCaT cells. RT-PCR analysis revealed that YM suppressed the expression of IL-$1{\beta}$ and MMP-9 however, it has no effects on the expression of TNF-${\alpha}$ and MMP-3. Our study suggests that Yukmijiwhang-tang exert protective actions on the UVB-irradiated HaCaT cells largely by anti-oxidative and anti-inflammatory processes.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.11
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• pp.5709-5714
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• 2012

Objective: To evaluate the effects of curcumin on matrixmetalloproteinase-9 (MMP-9) and invasion ability induced by transforming growth factor-${\beta}1$ (TGF-${\beta}1$) in MDA-MB-231 cells and potential mechanisms. Methods: Human breast cancer MDA-MB-231 cells were used with the CCK-8 assay to measure the cytotoxicity of curcumin. After treatment with 10 ng/ml TGF-${\beta}1$, with or without curcumin (${\leq}10{\mu}M$), cell invasion was checked by transwell chamber. The effects of curcumin on TGF-${\beta}1$-stimulated MMP-9 and phosphorylation of Smad2, extracellular-regulated kinase (ERK), and p38 mitogen activated protein kinases (p38MAPK) were examined by Western blotting. Supernatant liquid were collected to analyze the activity of MMP-9 via zymography. Following treatment with PD98059, a specific inhibitor of ERK, and SB203580, a specific inhibitor of p38MAPK, Western blotting and zymography were employed to examine MMP-9 expression and activity, respectively. Results: Low dose curcumin (${\leq}10{\mu}M$) did not show any obvious toxicity to the cells, while $0{\sim}10{\mu}mol/L$ caused a concentration-dependent reduction in cell invasion provoked by TGF-${\beta}1$. Curcumin also markedly inhibited TGF-${\beta}1$-regulated MMP-9 and activation of Smad2, ERK1/2 and p38 in a dose- and time-dependent manner. Additionally, PD98059, but not SB203580, showed a similar pattern of inhibition of MMP-9 expression. Conclusion: Curcumin inhibited TGF-${\beta}1$-stimulated MMP-9 and the invasive phenotype in MDA-MB-231 cells, possibly associated with TGF-${\beta}$/Smad and TGF-${\beta}$/ERK signaling.

• 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 the Korea Convergence Society
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• v.8 no.1
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• pp.107-114
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• 2017

To increase the bioavailability of Opuntia humifusa, O. humifusa was fermented using L. plantarum, and the characteristics of the fermented extract were confirmed. The characteristics of fermented extracts were investigated through viscosity, total polyphenol content, amino acid, antibacterial activity, cytokine analysis and zymography. The viscosity decreased by half and vitamin C remained almost unchanged during fermentation. Total polyphenols, most amino acids, total amino acids and major minerals were increased by fermentation. There was no cytotoxicity, and the activity of cytokines and MMP-9 in inflammation was inhibited. The inhibitory effect on inflammation and tissue destruction was found to be inhibited in L. plantarum fermentation extracts compared to hot water extracts. These results are expected to be the basis for the development of materials that enhance the functionality of L. plantarum-fermented O. humifusa extract.