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Herbal medicine In-Jin-Ho-Tang as a potential anti-cancer drug by induction of apoptosis in human hepatoma HepG2 cells. (사람 간암 세포주인 HepG2에 대한 인진호탕(茵陳蒿湯)의 항암 효과)

  • Yun, Hyun-Joung;Kim, Byung-Wan;Lee, Chang-Hyun;Jung, Jae-Ha;Heo, Sook-Kyung;Park, Won-Hwan;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.22 no.3
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    • pp.27-37
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    • 2007
  • Objectives: Hepatocellular carcinoma is the most common primary malignant tumor of the liver worldwide. In-Jin-Ho-Tang(IJHT) has been used as a traditional Chinese herbal medicine since ancient time. and today it is widely applied as a medication for jaundice which is associated with inflammation in liver. In this study, I investigated whether methanol extract of IJHT induced HepG2 cancer cell death. Methods: Cytotoxic activity of IJHT on HepG2 cells was using XTT assay. Apoptosis induction by Ros A in HCT116 cells was verified by the induction of cleavage of poly ADP-ribose polymerase (PARP). and activation of caspase-3, -8 and -9. The release of cytochrome c from mitochondria to cytosol. the level of Bcl-2 and Bax and the expression of p53 and p21 were examined by western blotting analysis. Furthermore, MAPKs activation was analyzed by western blotting analysis. Results: IJHT induced apoptosis in HepG2 cells. And treatment of IJHT resulted in the release of cytochrome c into cytosol, decreased anti-apoptotic Bcl-2, and increased pri-apoptotic Bax expression. IJHT markedly inactivated extracellular signal-regulated kinase (ERK1/2), and activated p38 mitogen-activated protein (MAP) kinase. Sodium orthovanadate (SOV), a phosphatase inhibitor, to reverse IJHT-induced ERK1/2 inactivation and SB203580, a specific p38 MAP Kinase inhibitor efficiently blocked apoptosis of HepG2. Thus, IJHT induces apoptosis in HepG2 cells via MAP kinase modulation. Conclusion: These results indicated that IJHT has some potential for use as an anti-cancer agent.

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The effect of rhBMP-2 on the osteoblastic differentiation of human periodontal ligament cells and gingival fibroblasts in vitro (치주인대세포와 치은섬유아세포의 분화에 미치는 rhBMP-2의 효과에 대한 연구)

  • Kim, Hyeon-Jong;Choi, Sang-Mook;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.32 no.2
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    • pp.389-402
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    • 2002
  • BMP can induce ectopic bone formation when implanted into sites such as rat muscle and can greatly enhance healing of bony defects when applied exogenously. In addition, BMP stimulated osteoblastic differentiation in vitro in various types of cells. The aim of this study was to investigate the effect of recombinant human bone morphogenetic protein(rhBMP-2) on the proliferation and osteoblastic differentiation of human periodontal ligament cells and gingival fibroblasts. The cell number and alkaline phosphatase activity were measured in 3 experimental groups of human periodontal ligament cells and gingival fibroblasts (control group, rhBMP-2 50ng/ml group, and rhBMP-2 100ng/ml group) at 1 and 2 weeks after culture. At the same time, total RNA of cultured cells were extracted and reverse trascription polymerase chain reaction(RT-PCR) was performed to determine the expression of mRNA of bone matrix protein. RhBMP-2 had no effect on the cell proliferation of human periodontal ligament cells and gingival fibroblasts. Alkaline phosphatase activity was elevated significantly by rhBMP-2 in both cells. And periodontal ligament cells showed significantly higher alkaline phosphatase activity than gingival fibroblasts. ${\beta}$-actin, type I collagen, alkaline phosphatase, BMP-2 mRNA were expressed in all of the samples. Osteopontin, osteocalcin mRNA were expressed in all periodontal ligament cell groups, and rhBMP-2 50ng/ml group, rhBMP-2 100ng/ml group of 2 week culture period of gingival fibroblasts. Bone sialoprotein mRNA was only expressed in rhBMP-2 50ng/ml group and rhBMP-2 100ng/ml group of 2-week culture period. These results suggest that rhBMP-2 stimulates osteoblastic differentiation in human periodontal ligament cells and gingival fibroblasts in vitro.

Effects of Poly-Gamma Glutamate Contents Cheonggukjang on Osteoblast Differentiation (폴리감마글루탐산(PGA) 함유량이 증가된 청국장이 조골세포 분화에 미치는 영향)

  • Lee, Ki Ho;Sim, Mi-Ok;Song, Yong Su;Jung, Ho Kyung;Jang, Ji-Hun;Kim, Min-Suk;Kim, Tae Mook;Lee, Hyo Eun;An, Byeong-Kwan;Jung, Won Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.5
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    • pp.664-670
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    • 2016
  • Cheonggukjang (CKJ) is a Korean traditional food made of fermented soybeans. In comparison to normal intake of soybeans, Cheonggukjang has high digestibility with bioactive, antioxidant substances, and thrombolytic enzymes. Recent studies have reported anti-oxidant, anti-cancer, anti-inflammatory, anti-obesity activities as well as inhibitory activities against osteoporosis for CKJ. In this study, we identified the effects of CKJ on osteoblast differentiation by increasing the polyglutamic acid (PGA) content of CKJ. Alkaline phosphatase (ALP) activity and mineralization significantly increased in response to treatment with both natural CKJ (CKJ A) and PGA-increased CKJ (CKJ B). However, CKJ B exhibited higher ALP activity and mineralization than CKJ A. Real-time reverse transcription PCR demonstrated that mRNA expression of osteoblastic-associated genes such as type I collagen, alkaline phosphatase, osteocalcin, and osteopontin in C2C12 cells was significantly up-regulated by CKJ A or B treatment. These results indicate that treatment with CKJ has an anabolic effect on bone by increasing osteoblastic differentiation and ALP activity. Increasing PGA content in CKJ had a greater effect than CKJ A on up-regulation of osteoblastic gene expression in osteoblast cells.

Cathepsin B in Eutopic and Ectopic Endometrial Tissues of Patients with Endometriosis

  • Kim, Chung-Hoon;Lee, You-Jeong;Kim, Jun-Bum;Lee, Young-Jin;Ahn, Jun-Woo;Kim, Sung-Hoon;Chae, Hee-Dong;Kang, Byung-Moon
    • Development and Reproduction
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    • v.17 no.2
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    • pp.133-140
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    • 2013
  • This study was performed to investigate the expression of cathepsin B mRNA and protein in eutopic and ectopic endometrial tissues of patients with endometriosis and in normal endometrial tissues and to clarify the association between the cathepsin B expression and endometriosis. A total of 40 women with histologically confirmed endometriosis were recruited for study group. For controls, 20 women undergoing operative treatment for uterine myoma, cervical intraepithelial neoplasia (CIN) or benign gynecologic conditions other than endometriosis were recruited. Eutopic endometrial tissues of both groups and ectopic endometrial tissue of study group were collected during the operations. We employed real time reverse transcriptase - polymerase chain reaction (RT-PCR) to quantify mRNA levels of cathepsin B in these tissues. Then, we performed western blot analysis to measure the protein levels of cathepsin B. The expressions of cathepsin B mRNA and protein were significantly higher in both eutopic and ectopic endometrial tissues of women with endometriosis than in endometrial tissues of controls. These data suggest that the higher expression of cathepsin B in the endometrial tissues might be associated with the development of endometriosis. In addition, eutopic endometrium itself with higher expression cathepsin B may play a pivotal role in the histogenesis of endometriosis.

Measurement of the Anti-oxidative Properties of Extract from Medicinal Plants Using an On-line HPLC-DPPH Assay (HPLC와 DPPH radical 소거능 측정 방법의 결합에 의한 약용 식물 추출물의 항산화 활성 비교)

  • Im, Do-Youn;Pyo, Byoung-Sik;Kim, Sun-Min;Lee, Kyoung-In
    • Journal of Life Science
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    • v.27 no.1
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    • pp.44-49
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    • 2017
  • Natural anti-oxidative compounds have important disease prevention and food preservation properties, in addition to anti-bacterial, anti-inflammation, anti-cancer, and skin whitening effects. High-performance liquid chromatography (HPLC), with an ultra vilolet (UV) detector coupled to a reverse phase C18 column and an online measurement system for 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radicals, was used to search for potent antioxidative compounds in crude extracts. The online HPLC-DPPH assay was then applied to confirm antioxidative compounds in water extracts from Radix of Pueraria lobata, Rhizoma of Zingiber officinale, Fructus of Chaenomeles sinensis, Cortex of Ulmus pumila, and Radix of Astragalus membranaceus. To determine the yields of the extracts, the Brix% of each extract solution was measured using a refractometer. When the relative DPPH radical scavenging ability values of the water extracts were compared with those of a positive control (ascorbic acid), the water extracts of P. lobata, C. sinensis, and U. pumila were 7.77%, 4.71%, and 4.19%, respectively. The results suggest that this method provides a useful assay for rapid measurement of DPPH radical scavenging abilities and conformation of antioxidative compounds in natural products. Moreover, it can reduce the time spent on the separation of active compounds from natural materials, such as medicinal plants, in addition to the use of reagents for separation.

Cucurbitacin-I, a Naturally Occurring Triterpenoid, Inhibits the CD44 Expression in Human Ovarian Cancer Cells (난소암 세포주의 CD44 발현에 미치는 Cucurbitacin-I의 효과)

  • Seo, Hee Won;Kim, Jin-Kyung
    • Journal of Life Science
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    • v.28 no.6
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    • pp.733-737
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    • 2018
  • Cucurbitacin-I, a natural triterpenoid derived from Cucurbitaceae family plants, exhibits a number of potentially useful pharmacological and biological activities. Indeed, the previous study demonstrated that cucurbitacin-I reduced the proliferation of colon cancer cells by enhancing apoptosis and causing cell cycle arrest at the G2/M phase. CD44, a type I transmembrane protein with the function of adhering to cells, mediates between the extracellular matrix and other cells through hyaluronic acid. Recent studies have demonstrated that an overexpression of the CD44 membrane receptor results in tumor initiation and growth, specific behaviors of cancer stem cells, the development of drug resistance, and metastasis. The aim was to examine the effect of cucurbitacin-I on CD44 expression human ovarian cancer cells because the effect of cucurbitacin-I on CD44 expression has not been reported. The expressions of CD44 mRNA and protein were detected using a quantitative real-time reverse-transcription polymerase chain reaction and a Western blot analysis, respectively. Treatment with cucurbitacin-I inhibited the expression of CD44 mRNA and protein. A subsequent analysis revealed that cucurbitacin-I blocked the phosphorylation of activator protein-1 (AP-1) and nuclear factor kappa-B ($NF-{\kappa}B$), which are key regulators of CD44 expression. Taken together, the data demonstrate that cucurbitacin-I regulates the AP-1 and $NF-{\kappa}B$ signaling pathways, leading to decreased CD44 expression.

A Study on Qualitative and Quantitative Analysis of Major ingredients in Scutellariae radix (황금(Scutellariae radix)의 주요 성분의 정성 및 정량분석에 관한 연구)

  • Rhee, Jae-Seong;Woo, Eun Ran;Kim, Nam-Hyuk;Lee, Eun-Ju;An, Duk-Kyun;Lee, Je-Hyun;Park, Seong Kyu;Park, Ho-Koon
    • Analytical Science and Technology
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    • v.10 no.2
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    • pp.91-104
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    • 1997
  • Scutellariae radix has been used on the control of body fever as oriental medicine for thousand years. Analytical aspect for the main components of Scutellariae radix was set up improving sensitivity and resolution. The analysis of 3 different flavonoids present in Scutellariae radix-baicalin, baicalein, wogonin-was conducted by means of high performance liquid chromatography with ODS reverse phase column in conjunction with a Photo Diode Array UV detector(280nm) at $40^{\circ}C$. Mobile phase was carried out at 1mL/min, composed of acetonitrile and 0.1M phosphoric acid in the form of a gradient method. Under these circumstances the retention time for baicalin, baicalein, wogonin was 7.65, 11.65 and 14.12 minutes respectively. As a result for the efficiency on extraction of active ingredients with proposed analytical process according to it's growing districts, Sunchang in Junbuk for baicalin and Bulkyo in, Junnam for bicalein and wogonin have shown the best results. Even the extraction at room temperature was satisfactory. Among acids, 0.1M acetic acid revealed the best achievements. The mixture of acetonitrile and 0.2M phosphoric acid(75:25) has been shown the best efficiency as well as stability for the extraction of active ingredients.

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Compression and Decompression Properties of Some Vegetables (몇가지 채소류의 압축 및 비압축 특성)

  • Min, Young-Kyoo;Jeong, Heon-Sang
    • Korean Journal of Food Science and Technology
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    • v.29 no.2
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    • pp.266-272
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    • 1997
  • In order to investigate the compression and decompression properties of cucumber, radish, garlic, ginger and potato, edible parts of samples were prepared to size of ${\Phi}\;5\;mm{\times}H\;5\;mm$, and force deformation relationship during application and removal of force were observed. Compositions of sample and cell characteristics were measured, and correlations between them were investigated. Deformation rate was large in initial stage of compression and decreased afterward, but the reverse trends were observed in the decompression. The time and deformation to 9 N were large of 5.30 sec and 1.344 mm in potato, and small of 4.62 sec and 0.896 mm in garlic, respectively. Force(y)-deformation(x) curve between compression and decompression were clearly showed hysteresis loop and relationship of x and y were as follows: y=esp (a+b log(x)). The maximum work was $3.888{\sim}5.099{\times}10^{-3}\;J$ for potato in compression and $2.09{\times}10^{-3}\;J$ for garlic in decompression. Irrecoverable work were large as $77{\sim}96%$ in cucumber, radish and potato, and small as $36{\sim}42%$ in garlic. Compression deformation were large as $1.016{\sim}1.344\;mm$ in potato, and small as $0.656{\sim}0.896\;mm$ in garlic. Degree of elasticity were large as $0.756{\sim}0.777$ in garlic, and small as $0.301{\sim}0.465$ in radish and potato. Compression and decompression characteristic values were showed high correlation with moisture, viscosity of juice, ceil size, density and regularity.

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Fenofibrate decreases radiation sensitivity via peroxisome proliferator-activated receptor ${\alpha}$-mediated superoxide dismutase induction in HeLa cells

  • Liu, Xianguang;Jang, Seong-Soon;An, Zhengzhe;Song, Hye-Jin;Kim, Won-Dong;Yu, Jae-Ran;Park, Woo-Yoon
    • Radiation Oncology Journal
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    • v.30 no.2
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    • pp.88-95
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    • 2012
  • Purpose: The fibrates are ligands for peroxisome proliferator-activated receptor (PPAR) ${\alpha}$ and used clinically as hypolipidemic drugs. The fibrates are known to cause peroxisome proliferation, enhance superoxide dismutase (SOD) expression and catalase activity. The antioxidant actions of the fibrates may modify radiation sensitivity. Here, we investigated the change of the radiation sensitivity in two cervix cancer cell lines in combination with fenofibrate (FF). Materials and Methods: Activity and protein expression of SOD were measured according to the concentration of FF. The mRNA expressions were measured by using real time reverse-transcription polymerase chain reaction. Combined cytotoxic effect of FF and radiation was measured by using clonogenic assay. Results: In HeLa cells total SOD activity was increased with increasing FF doses up to 30 ${\mu}M$. In the other hand, the catalase activity was increased a little. As with activity the protein expression of SOD1 and SOD2 was increased with increasing doses of FF. The mRNAs of SOD1, SOD2, $PPAR{\alpha}$ and $PPAR{\gamma}$ were increased with increasing doses of FF. The reactive oxygen species (ROS) produced by radiation was decreased by preincubation with FF. The surviving fractions (SF) by combining FF and radiation was higher than those of radiation alone. In Me180 cells SOD and catalase activity were not increased with FF. Also, the mRNAs of SOD1, SOD2, and $PPAR{\alpha}$ were not increased with FF. However, the mRNA of $PPAR{\gamma}$ was increased with FF. Conclusion: FF can reduce radiation sensitivity by ROS scavenging via SOD induction in HeLa. SOD induction by FF is related with $PPAR{\alpha}$.

Inactivation of Brain myo-Inositol Monophosphate Phosphatase by Pyridoxal-5'-Phosphate

  • Kim, Dae-Won;Hong, Joung-Woo;Eum, Won-Sik;Choi, Hee-Soon;Choi, Soo-Hyun;Kim, So-Young;Lee, Byung-Ryong;An, Jae-Jin;Lee, Sun-Hwa;Lee, Seung-Ree;Kwon, Oh-Shin;Kwon, Hyeok-Yil;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Choi, Soo-Young
    • BMB Reports
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    • v.38 no.1
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    • pp.58-64
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    • 2005
  • Myo-inositol monophosphate phosphatase (IMPP) is a key enzyme in the phosphoinositide cell-signaling system. This study found that incubating the IMPP from a porcine brain with pyridoxal-5'-phosphate (PLP) resulted in a time-dependent enzymatic inactivation. Spectral evidence showed that the inactivation proceeds via the formation of a Schiff's base with the amino groups of the enzyme. After the sodium borohydride reduction of the inactivated enzyme, it was observed that 1.8 mol phosphopyridoxyl residues per mole of the enzyme dimer were incorporated. The substrate, myo-inositol-1-phosphate, protected the enzyme against inactivation by PLP. After tryptic digestion of the enzyme modified with PLP, a radioactive peptide absorbing at 210 nm was isolated by reverse-phase HPLC. Amino acid sequencing of the peptide identified a portion of the PLP-binding site as being the region containing the sequence L-Q-V-S-Q-Q-E-D-I-T-X, where X indicates that phenylthiohydantoin amino acid could not be assigned. However, the result of amino acid composition of the peptide indicated that the missing residue could be designated as a phosphopyridoxyl lysine. This suggests that the catalytic function of IMPP is modulated by the binding of PLP to a specific lysyl residue at or near its substrate-binding site of the protein.