• Title/Summary/Keyword: MMP activity

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Antioxidant, Anti-inflammation, and Anti-aging Effects of Ethyl Acetate Fraction from Cynanchum paniculatum Extracts (산해박 분획물의 항산화, 항염증 및 항노화 활성 효과 연구)

  • Jae Yong Seo;Go Eun Kim;Ji Soo Ryu;A Reum Jang;Su An Sim;Jung No Lee;Seunghee Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.1
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    • pp.85-93
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    • 2024
  • In this study, we prepared fractions from Cynanchum paniculatum (C. paniculatum) with ethyl acetate and identified the structure and content of paeonol, an indicator substance. Fractions were obtained by sequential solvent fractions using n-hexane, methylene chloride, ethyl acetate, n-butanol, and distilled water, and showed the highest paeonol content in ethyl acetate (EtOAc). The structure of the ethyl acetate fraction (EA) was confirmed by nuclear magnetic resonance (NMR) and the content was determined by high performance liquid chromatography (HPLC) analysis. The antioxidant activity of EA was tested through its ability to scavenge 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and its ability to inhibit nitric oxide (NO) production in RAW 264.7 cells induced by lipopolysaccharide (LPS) was tested for its ability to inhibit NO production in a concentration-dependent manner. Furthermore, MMP-1 production induced by ultraviolet B (UVB) was decreased in a concentration-dependent manner by the EA, and the ability to produce procollagen type-I was increased in a concentration-dependent manner. In addition, as a result of conducting clinical trials using cosmetic formulations containing EA, the effect of improving wrinkles around the eyes was confirmed. Thus, it is expected that Cynanchum paniculatum can be used as a natural functional cosmetic material with antioxidant and anti-aging effects.

PDI-like Enzyme in Human Follicular Fluid Converts 72 kDa Gelatinase into GA110 (사람 난포액에 존재하는 72 kDa Geletinase로부터 GA110을 만드는 PDI-like PDI-like Enzyme)

  • Kim Jisoo;Kim Haekwon
    • Development and Reproduction
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    • v.7 no.2
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    • pp.105-112
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    • 2003
  • Previously, we discovered a new MMP-2 isoform GA110, of which appearance in human follicular fluid(FF) and serum was increased by EDTA. The present study was conducted to investigate how GAI 10 can appear by EDTA. To examine possible involvement of protein disulfide isomerase(PDI), an enzyme responsible for the dimerization of protein via disulfide formation, effect of PDI inhibitor on the appearance of GA110 by EDTA was investigated. When PDI inhibitor added to FF before EDTA treatment, the gelatinolytic activity of GA110 was abolished in a concentration dependent manner. By contrast, the activity of 72 kDa gelatinase increased. However, the PDI inhibitor added to FF after EDTA treatment, the gelatinolytic activity of GA110 was unaffected. To find out the nature of the enzyme which converts 72 kDa gelatinase into GAI 10, chromatographic separation method of FF proteins was done. Using hydroxyapatite column, fractions rich in 72 kDa gelatinase were isolated and pooled. By using this pool as substrate for the 72 kDa converting enzyme, protein fractions containing the converting activity were obtained from chromatographic separation of FF onto glutathione sepharose fast flow column. When immunoblotting was performed on this enzymatically active protein fractions against polyclonal anti-PDI antibody, distinct immunoreactivity was observed, although appeared in smaller molecular weight region. Based on these observations, it is suggested that the appearance of GAI 10 in FF by EDTA treatment could be due to an activation of PDI-like enzyme, which dimerizes 72 kDa gelatinase into GAI 10 via the formation of disulfide bond between molecules.

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Mechanism Underlying a Proteasome Inhibitor, Lactacystin-Induced Apoptosis on SCC25 Human Tongue Squamous Cell Carcinoma Cells (사람혀편평상피세포암종세포에서 proteasome 억제제인 lactacystin에 의해 유도된 세포자멸사의 기전에 대한 연구)

  • Baek, Chul-Jung;Kim, Gyoo-Cheon;Kim, In-Ryoung;Lee, Seung-Eun;Kwak, Hyun-Ho;Park, Bong-Soo;Tae, Il-Ho;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.261-276
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    • 2009
  • Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

Protective effect of Gabjubaekmok (Diospyros kaki) extract against amyloid beta (Aβ)-induced cognitive impairment in a mouse model (아밀로이드 베타(amyloid beta)로 유도된 인지장애 마우스 모델에서 갑주백목(Diospyros kaki) 추출물의 인지기능 및 뇌 신경세포 보호 효과)

  • Yoo, Seul Ki;Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Han, Hye Ju;Park, Hyo Won;Kim, Chul-Woo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.379-392
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    • 2019
  • The current study investigated the effect of Gabjubaekmok (Diospyros kaki) ethanolic extract (GEE) on $H_2O_2$-induced human neuroblastoma MC-IXC cells and amyloid beta $(A{\beta})_{1-42}$-induced ICR (Institute of Cancer Research) mice. GEE showed significant antioxidant activity that was evaluated based on ABTS, DPPH scavenging activity, and inhibition of malondialdehyde (MDA) and acetylcholinesterase activity. Further, GEE inhibited ROS production and increased cell viability in $H_2O_2$-induced MC-IXC cells. Administration of GEE ameliorated the cognitive dysfunction on $A{\beta}$-induced ICR mice as evaluated using Y-maze, passive avoidance, and Morris water maze tests. Results of ex vivo test using brain tissues showed that, GEE protected the cholinergic system and mitochondrial functions by increasing the levels of antioxidants such as ROS, mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) against $A{\beta}$-induced cognitive dysfunction. Moreover, GEE decreasd the expression levels of apoptosis-related proteins such as $TNF-{\alpha}$, p-JNK, p-tau, BAX and caspase 3. While, expression levels of p-Akt and $p-GSK3{\beta}$ increased than $A{\beta}$ group. Finally, gallic acid was identified as the main compound of GEE using high performance liquid chromatography.

Skin Protection Effect of Grape Pruning Stem Extract on UVB-induced Connective Tissue Injury (포도전정가지 추출물이 UVB로 유도된 결합 조직 손상에 미치는 피부 보호 효과)

  • Kim, Joung-Hee;Kim, Keuk-Jun
    • Journal of Life Science
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    • v.28 no.2
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    • pp.141-147
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    • 2018
  • This study aimed to analyze the contents of rutin, procyanidin B3, quercetin, and kaempferol, known to have antioxidant, anti-inflammatory, and anti-carcinogenic effects, among the polyphenol types contained in grape pruning stem extracts (GPSE). It utilized grape stems discarded after harvest to measure the effects of GPSE on skin moisture, inhibition of skin cell proliferation, and anti-inflammatory activity on the damaged skin of HR-1 mice induced with ultraviolet B (UVB), and to verify the applicability of GPSE as a material for functional food and functional cosmetics. The polyphenol was extracted from grape pruning stems with 80% EtOH, and then the extract was used while storing at $-20^{\circ}C$, after filtering, concentrating, and freeze-drying it. The content of an active ingredient of GPSE was analyzed using high performance liquid chromatography (HPLC). From 53 kg of the grape pruning stem specimen, 2.34 kg of the EtOH fraction extracts were extracted to achieve a 4.42% yield ratio. Analysis of the active ingredients showed 0.28 mg/g of procyanidin B3, 12.81 mg/g of rutin, 0.51 mg/g of quercetin, and 8.24 mg/g of kaempferol. After UVB irradiation on the dermis, to confirm the degree of inhibition of collagen synthesis, we examined the protein expression of MMP-9 using immunohistochemical staining. The results of this study confirm the existence of active polyphenol types, such as rutin, kaempferol, quercetin, and procyanidin B3, in GPSE. Moreover, the study found that GPSE has anti-collagenase effects and it decreases the effects of UV damage on skin barrier function. GPSE is a functional ingredient with a potential for skin protection effects, and it has high utilization potential as an ingredient for functional cosmetics.

Functional screening of Asparagus officinalis L. stem and root extracts (아스파라거스 줄기 및 뿌리 추출물의 기능성 검증)

  • Han, Joon-Hee;Hong, Min;Lee, Jaehak;Choi, Da-Hye;Lee, Sun-Yeop;Kwon, Tae-Hyung;Lee, Jae-Hee;Lee, Yong-Jin;Yu, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.53 no.1
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    • pp.46-54
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    • 2021
  • The biological activities of non-edible extracts of asparagus stems and roots were investigated using hot water and ethanol. The highest contents of rutin and total polyphenol were 31.74 mg/g and 20.14 mg GAE/g, respectively, in the stem hot water extract. ABTS and DPPH radical scavenging activities were 541.1±21.0 and 649.5±6.6 ㎍/mL, respectively, in stem hot water extract. All extracts were non-cytotoxic in HepG2 cells, but 200 ㎍/mL stem extracts tended to decrease the viability of RAW 264.7 cells. The highest xanthine oxidase inhibitory activity was 43.68% in the root hot water extract at 200 ㎍/mL. The expression level of MMP-9 was significantly decreased in the asparagus extracts. The highest GGT, AST, and LDH activities showed a concentration-dependent decrease in the stem ethanol extract. In conclusion, the presence of bioactive substances in the non-edible extracts of asparagus was confirmed for the development of extracts with antioxidant, hepatoprotective and anti-gout activities.

A STUDY ON THE ACTIVITY OF PROTEINASE AND PROTEINASE INHIBITOR IN PULPAL AND PERIAPICAL PATHOSES (치수 및 치근단 질환에서의 단백분해효소 및 단백분해효소 억제제의 활성도에 관한 연구)

  • Kim, Jin-Woo;Lee, Suk-Keun;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.25 no.4
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    • pp.509-526
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    • 2000
  • It is known that injuries to the dentin have a corresponding inflammatory effect on the pulp and these inflammatory effects frequently result in pulpal pathoses due to progressive degradation of pulpal connective tissue. It was supposed that the tissue degradation in different inflammatory process was controlled by proteinase activity and antiproteinase activity. Therefore, the purpose of this study was to examine the pulp and periapical pathoses in terms of the activities of proteinase and proteinase inhibitor, 37 pulpal tissues were divided by clinical diagnostic criteria into normal pulp, acute inflamed pulp, and chronic inflamed pulp, and then those groups were subdivided by histopathological findings into 5 pulpal pathoses groups, i.e. normal pulp (P1, n=8), chronic pulpitis with fibrotic change (P2, n=2), chronic pulpitis with dystrophic calcification (P3, n=11), chronic pulpitis with pulp abscess (P4, n=7), acute pulpitis with necrotic change (P5, n=4), 26 periapical tissues were also divided by ordinary histopathological findings into 3 periapical pathoses group, i.e., granuloma (A1, n=17), cyst (A2, n=2) and abscess (A3, n=7). The activities of proteinases (cathepsin G, MMP-3) and proteinase inhibitors (${\alpha}1$-AT, TIMP-1 and, SLPI) were evaluated by RT-PCR and immunohistochemical methods. The results were as follows. 1. Generally, the intensity of immunohistochemical staining of proteinases and proteinase inhibitors increased in P2 and P5 groups compared to P1 group. 2. The immunohistochemical stain of proteinases and proteinase inhibitors was intensely detected in P2 group, showing low inflammatory reaction and low tissue degradation, but it was reduced in P3 and P4 groups, showing severe tissue degradation. 3. The distribution of proteinases and proteinase inhibitors in pulpal pathoses was consistently presented by immunohistochemical staining, while the expression of proteinase and/or proteinase inhibitors mRNAs in pulpal pathoses was occasionally detected by RT-PCR methods. 4. RT-PCR of proteinase and proteinase inhibitors was usually positive in P2, showing rare tissue degradation, but it was almost negative in P3 and P4, showing severe tissue degradation. 5. We presume that the reason why the level of proteinase and proteinase inhibitors was so sparse in RT-PCR method is due to the abrupt decrease of mRNA synthesis or degradation of synthesized mRNA of proteinase and/or proteinase inhibitors depend on the inflammatory reaction and/or on the degradation of pulp tissues(P3, P4). 6. Pulpal pathoses groups showed significant lower RT-PCR detection of proteinases and proteinase inhibitors than the periapical pathoses group(p<0.05), and there is no significant difference among the periapical pathoses groups(p>0.05).

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Anti-skin Aging Potential of Alcoholic Extract of Phragmites communis Rhizome

  • Ha, Chang Woo;Kim, Sung Hyeok;Lee, Sung Ryul;Jang, Sohee;Namkoong, Seung;Hong, Sungsil;Lim, Hyosun;Kim, Youn Kyu;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
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    • v.33 no.6
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    • pp.604-614
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    • 2020
  • Chronological aging and photoaging affect appearance, causing wrinkles, pigmentation, texture changes, and loss of elasticity in the skin. Phragmites communis is a tall perennial herb used for its high nutritional value and for medicinal purposes, such as relief from fever and vomiting and facilitation of diuresis. In this study, we investigated the effects of ethanol extract of P. communis rhizome (PCE) on skin aging. The total flavonoid and total phenolic content in PCE were 2.92 ± 0.007 ㎍ of quercetin equivalents (QE) and 231.8 ± 0.001 ㎍ of gallic acid equivalents (GAE) per 100 mg of dried extract (n = 3). The half-maximal inhibitory concentration (IC50) values of PCE for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and hydrogen peroxide scavenging activities were 0.96 and 0.97 mg/mL, respectively. PCE showed inhibitory effects on tyrosinase when L-tyrosine (IC50 = 1.25 mg/mL) and L-3,4-dihydroxyphenylalanine (IC50 = 0.92 mg/mL) were used as substrates. PCE treatment up to 200 ㎍/mL for 24 h did not cause any significant cytotoxicity in B16F10 melanocytes, human dermal fibroblasts (HDFs), and HaCaT keratinocytes. In B16F10 melanocytes, PCE (25 and 50 ㎍ /mL) inhibited melanin production and cellular tyrosinase activity after challenge with α-melanocyte-stimulating hormone (α-MSH; p < 0.05). In HDFs, PCE suppressed the mRNA expression of matrix metalloproteinase-1 (MMP-1) and reduced the activity of elastase (p < 0.05). In addition, ultraviolet B (UVB)-mediated downregulation of hyaluronic acid synthase-2 gene expression in HaCaT keratinocytes was also effectively suppressed by PCE treatment. Overall, our results showed that PCE has potential anti-skin aging activity associated with the suppression of hyperpigmentation, wrinkle formation, and reduction in dryness. PCE is a promising candidate for the development of an anti-skin aging cosmetic ingredient.

Association of a Methanol Extract of Rheum undulatum L. Mediated Cell Death in AGS Cells with an Intrinsic Apoptotic Pathway

  • Hong, Noo Ri;Park, Hyun Soo;Ahn, Tae Seok;Jung, Myeong Ho;Kim, Byung Joo
    • Journal of Pharmacopuncture
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    • v.18 no.2
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    • pp.26-32
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    • 2015
  • Objectives: Rheum undulatum L. has traditionally been used for the treatment of many diseases in Asia. However, its anti-proliferative activity in cancer has still not been studied. In the present study, we investigated the anti-cancer effects of methanol extract of Rheum undulatum L. (MERL) on human adenocarcinoma gastric cell lines (AGS). Methods: To investigate the anti-cancer effect of MERL on AGS cells, we treated the AGS cells with varying concentrations of MERL and performed 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. Cell cycle analyses, measurements of the mitochondrial membrane potential (MMP), caspase activity assays and Western blots were conducted to determine whether AGS cell death occurred by apoptosis. Results: Treatment with MERL significantly inhibited growth of AGS cells in a concentration dependent manner. MERL treatment in AGS cells leaded to increased accumulation of apoptotic sub G1 phase cells in a concentration dependent manner. In control cultures, 5.38% of the cells were in the sub G1 phase. In MERL treated cells, however, this percentage was significantly increased (9.95% at $70{\mu}g/mL$, 15.94% at $140{\mu}g/mL$, 26.56% at $210{\mu}g/mL$ and 38.08% at $280{\mu}g/mL$). MERL treatment induced the decreased expression of pro-caspase-8 and -9 in a concentration dependent manner, whereas the expression of the active form of caspase-3 was increased. A subsequent Western blot analysis revealed increased cleaved levels of poly (ADP-ribose) polymerase (PARP) protein. Also, treatment with MERL increased the activities of caspase-3 and -9 compared with the control. MERL treatment increased the levels of the pro-apoptotic truncated Bid (tBid) and Bcl2 Antagonist X (Bax) proteins and decreased the levels of the anti-apoptotic B-cell lymphoma 2 (Bcl-2) protein, whose is the stabilization of mitochondria. However, inhibitions of p38, extracellular signal regulated kinases (ERKs) and C-Jun N-terminal kinases (JNK) by MERL treatment did not affect cell death. Conclusion: These results suggest that MERL mediated cell death is associated with an intrinsic apoptotic pathway in AGS cells.

Baicalein Inhibits the Migration and Invasion of B16F10 Mouse Melanoma Cells through Inactivation of the PI3K/Akt Signaling Pathway

  • Choi, Eun-Ok;Cho, Eun-Ju;Jeong, Jin-Woo;Park, Cheol;Hong, Su-Hyun;Hwang, Hye-Jin;Moon, Sung-Kwon;Son, Chang Gue;Kim, Wun-Jae;Choi, Yung Hyun
    • Biomolecules & Therapeutics
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    • v.25 no.2
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    • pp.213-221
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    • 2017
  • Baicalein, a natural flavonoid obtained from the rhizome of Scutellaria baicalensis Georgi, has been reported to have anticancer activities in several human cancer cell lines. However, its antimetastatic effects and associated mechanisms in melanoma cells have not been extensively studied. The current study examined the effects of baicalein on cell motility and anti-invasive activity using mouse melanoma B16F10 cells. Within the noncytotoxic concentration range, baicalein significantly inhibited the cell motility and invasiveness of B16F10 cells in a concentration-dependent manner. Baicalein also reduced the activity and expression of matrix metalloproteinase (MMP)-2 and -9; however, the levels of tissue inhibitor of metalloproteinase-1 and -2 were concomitantly increased. The inhibitory effects of baicalein on cell motility and invasiveness were found to be associated with its tightening of tight junction (TJ), which was demonstrated by an increase in transepithelial electrical resistance and downregulation of the claudin family of proteins. Additionally, treatment with baicalein markedly reduced the expression levels of lipopolysaccharide-induced phosphorylated Akt and the invasive activity in B16F10 cells. Taken together, these results suggest that baicalein inhibits B16F10 melanoma cell migration and invasion by reducing the expression of MMPs and tightening TJ through the suppression of claudin expression, possibly in association with a suppression of the phosphoinositide 3-kinase/Akt signaling pathway.