• Title/Summary/Keyword: RNA 추출

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Induction of Apoptosis in AGS Human Gastric Cancer Cells by Platycarya strobilacea Leaf Extract (굴피나무 잎 추출물의 위암세포에 대한 세포사멸 유도 효과)

  • Lee, Hyeong-Seon
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.283-288
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    • 2021
  • This study investigated the anticancer activity of methanol extract from Platycarya strobilacea leaf in AGS human gastric cancer cells. We determined the cell viability effect of P. strobilacea using MTS assay. Apoptosis induction and cell cycle arrest were confirmed by fluorescein isothiocyanate and propidium iodide staining using cellometer K2. The mRNA expression levels of the Bcl-2 family were confirmed by reverse transcription-polymerase chain reaction. The cell viability was decreased in a dose-dependent manner treated with different concentrations of P. strobilacea. Total, early, and late apoptotic cells were dramatically increased, and the cell cycle was arrested at the sub-G1 phase. The mRNA expressions of Bcl-2 and Bcl-xL were reduced, whereas pro-apoptotic factors, Bax and Bak, were increased in a dose-dependent manner. These results suggested that P. strobilacea leaf extract induced significant apoptotic activity through an intrinsic mitochondria pathway.

Effects of Adenophora triphylla Ethylacetate Extract on mRNA Levels of Antioxidant Enzymes in Human HepG2 Cells (인간 HepG2 Cell에서 항산화 효소의 mRNA 발현에 대한 잔대 에틸아세테이트 추출물 효과)

  • Choi, Hyun-Jin;Kim, Soo-Hyun;Oh, Hyun-Taek;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1238-1243
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    • 2008
  • The root of Adenophora triphylla is widely used as traditional herbal medicine in Korea. We studied its effects on sodium nitroprusside (SNP) cytotoxicity and antioxidant genes expression in HepG2 cells. To study whether Adenophora triphylla ethylacetate extract (ATea) inhibited NO-induced cell death, HepG2 cells were preincubated for 24 hr with 50 and 100 $\mu$g/mL ATea followed by 24-hr exposure to 0.5 mM SNP (exogenous NO donor). No-induced cytotoxicity was inhibited by pretreatment of ATea, as assessed by mitochondrial dehydrogenase activity (MTT assay). We further investigated the effects of ATea on mRNA levels of various enzymes of the antioxidant system such as Cu, Zn superoxide dismutase (SOD 1), Mn SOD (SOD 2), glutathione peroxidase (GPx), catalase and several enzymes of the glutathione metabolism [glutathione reductase (GR), $\gamma$-glutamyl-cystein synthetase (GCS), glutathione-S-transferase (GST), $\gamma$-glutamyltranspeptidase ($\gamma$-GT), glucose-6-phosphate dehydrogenase (G6PD)] by RT-PCR. CAT, GCS, GR and G6PD mRNA levels were increased after treatment with ATea. The SOD 1, SOD 2, GPx, GST and $\gamma$-GT mRNA levels were not affected in ATea-treated HepG2 cells. We concluded that ATea have an indirect antioxidant effects, perhaps via induction of CAT, GCS, GR and G6PD.

Effects of Sasa quelpaertensis Extract on mRNA and microRNA Profiles of SNU-16 Human Gastric Cancer Cells (SNU-16 위암 세포의 mRNA 및 miRNA 프로파일에 미치는 제주조릿대 추출물의 영향)

  • Jang, Mi Gyeong;Ko, Hee Chul;Kim, Se-Jae
    • Journal of Life Science
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    • v.30 no.6
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    • pp.501-512
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    • 2020
  • Sasa quelpaertensis Nakai leaf has been used as a folk medicine for the treatment of gastric ulcer, dipsosis, and hematemesis based on its anti-inflammatory, antipyretic, and diuretic characteristics. We have previously reported the procedure for deriving a phytochemical-rich extract (PRE) from S. quelpaertensis and how PRE and its ethyl acetate fraction (EPRE) exhibits an anticancer effect by inducing apoptosis in various gastric cancer cells. To explore the molecular targets involved in this apoptosis, we investigated the mRNA and microRNA profiles of EPRE-treated SNU-16 human gastric cancer cells. In total, 2,875 differentially expressed genes were identified by RNA sequencing, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that the EPRE-modulated genes are associated with apoptosis, mitogen-activated protein kinase, inflammatory response, tumor necrosis factor signaling, and cancer pathways. Subsequently, protein-protein interaction network analysis confirmed interactions among genes associated with cell death and apoptosis, and 27 differentially expressed microRNAs were identified by further sequencing. Here, GO and KEGG pathway analysis revealed that EPRE modified the expression of microRNAs associated with the cell cycle and cell death, as well as signaling of tropomyosin-receptor-kinase receptor, transforming growth factor-b, nuclear factor kB, and cancer pathways. Taken together, these results provide insight into the mechanisms underlying the anticancer effect of EPRE.

Effect of Inonotus obliquus Extract on the Expression MMPs and HAS-2 (차가버섯 추출물의 MMPs 발현 저해 및 HAS-2 발현 증가효과)

  • Lee, So-Hun;Park, Sun-Hee;Lee, Kang-Hyuk;Park, Soo-Jin;Kim, Young-Heui
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.3
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    • pp.237-245
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    • 2011
  • In order to investigate the possibility of Inonotus obliquus extract as an active ingredient for wrinkle-care cosmetics, we prepared 70 % ethanolic extract of Inonotus obliquus and measured its DPPH radical scavenging activity and elastase inhibitory activity. We also evaluated the effect of Inonotus obliquus extract on expression of MMPs and HAS-2 in fibroblast and HaCaT cell, respectively. Inonotus Obliquus extract showed DPPH radical scavenging activity and elastase inhibitory activity in a dose-dependant manner ($SC_{50}$ = 91 ${\mu}g$/mL, $IC_{50}$=124 ${\mu}g$/mL). Inonotus obliquus extract inhibited the expression of MMP-1 mRNA 50 ~ 79 % at concentrations of 5 ~ 25 ${\mu}g$/mL, and reduced the expression of MMP-2 around 20 % at a concentration of 10 ${\mu}g$/mL. And it also reduced the expression of MMP-9 54 ~ 70 % in tconcentrations of 5 to 25 ${\mu}g$/mL. Furthermore, Inonotus obliquus extract increased HAS-2 mRNA expression in a dose-dependent manner in concentrations of 5 to 25 ${\mu}g$/mL without cytotoxicity. These results suggested that Inonotus Obliquus extract could be useful as an active ingredient for wrinkle-care cosmetics.

Effect of hot water and ethanol extracts from Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production in adipogenesis of 3T3-L1 cells (3T3-L1 세포분화 중 지방축적 및 ROS 생성에 대한 연화 열수 및 에탄올 추출물의 효과)

  • Oh, Ji-Won;Lee, Jin-Ha;Lee, Ok-Hwan;Kim, Kye-Hoon;Kim, Hey-Ran;Lee, Hyo-Ku
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.744-750
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    • 2015
  • The present study was designed to investigate the effects of hot water and ethanol extracts of Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with both hot water and ethanol extracts for up to 8 days following standard induction of differentiation. Regarding anti-adipogenic activity, compared with the control, the hot water and ethanol extracts significantly inhibited lipid accumulation (37.4 and 66.6%, respectively) and ROS production (46.4 and 46.8%, respectively) during adipogenesis in 3T3-L1 cells. Treatment with hot water and ethanol extracts significantly inhibited mRNA expression of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), thereby reducing the mRNA expression of adipocyte-specific fatty acid binding protein (aP2). Moreover, both the extracts significantly inhibited mRNA expression of NADPH oxidase (NOX4). Overall, our research suggests that N. nucifera Gaertner flower extracts might be a valuable source of bioactive compounds that exhibit anti-adipogenic activity and could have applications in the field of medicine and food industry.

Inhibitory Effects of Allium senescens L. Methanol Extracts on Reactive Oxygen Species Production and Lipid Accumulation during Differentiation in 3T3-L1 Cells (두메부추(Allium senescens L.) 메탄올 추출물의 지방세포 내 활성산소종 생성 및 지질축적 억제 효능)

  • Choi, Hye-Young;Kim, Gun-Hee
    • Korean Journal of Food Science and Technology
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    • v.46 no.4
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    • pp.498-504
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    • 2014
  • Allium senescens L. is perennial plant of the Liliaceae family that grows throughout Korea. In this study, we investigated the effect of Allium senescens L. methanol extracts on reactive oxygen species (ROS) production and lipid accumulation during adipogenesis. Our results indicated that 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of Allium senescens L. methanol extracts increased in a dose-dependent manner. Allium senescens L. methanol extracts suppressed ROS production and lipid accumulation during adipogenesis. In addition, Allium senescens L. methanol extracts inhibited the mRNA expression of the pro-oxidant enzyme, such as G6PDH and lead to a reduction in the mRNA levels of the transcription factors, such as sterol regulatory element binding proteins 1c, peroxisome proliferator-activated receptor ${\gamma}$, and CCAAT/enhancer-binding proteins ${\alpha}$. These results indicate that Allium senescens L. methanol extracts inhibit adipogenesis by modulating ROS production associated with ROS-regulating genes and directly down-regulating adipogenic transcription factors.

Increase in 3-Hydroxy-3-Methylglutarly Coenzyme A Reductase mRNA Level in Tomato by Fungal Elicitors and Mechanical Wounding (Fungal Elicitor와 기계적 상해에 의한 토마토 HMGR mRNA 증가)

  • 박희성;이용세
    • Korean Journal Plant Pathology
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    • v.12 no.3
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    • pp.285-290
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    • 1996
  • 3-hydroxy-3-methyglutaryl coenzyme A reductase (HMGR)는 phytoalexin을 포함하는 수 많은 isoprenoid화합물의 생합성을 조절하는 효소이다. 토마토의 경우 sesquiterpenoid phytoalexin류가 식물방어를 위한 반응산물로서 축적되는 것이 알려져 있다. Verticil-lium albo-atrum이나 Fusarium oxysporum으로부터 추출한 elicitor를 토마토의 배양세포에 처리하는 경우 처리량의 증가에 따른 2.7kb 크기의 HMGR mRNA의 상당한 유도증가가 토마토의 HMG2 DNA를 이용한 northern hybridization에 의해 관찰되었다. 토마토의 잎, 뿌리, 줄기 등에 기계적 상해를 가하는 경우에서도 HMGR mRNA는 2단계를 걸쳐 증가함이 관찰되었다. HMGR mRNA는 양 실험의 경우 모두 9시간에서 12시간 사이에서 최대발현됨이 관찰되었다.

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Anti-melanogenesis Effect of Canavalia lineata Extract (해녀콩(Canavalia lineata THUNB. DC.) 추출물의 멜라닌 생성 억제 효과)

  • Bu Hee-Jung;Riu Key-Zung;Lee Sunjoo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.485-489
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    • 2004
  • Melanin pigmentation in human skin is a major defensive mechanism against ultraviolet light of the sun. Tyrosinase plays a key role in the biosynthesis of melanin. This is why many researches have been focused on regulations in controlling the epidermal melanization. We found that extract of Canavalia lineata inhibits mushroom tyrosinase activity, dopa oxidase activity, and melanin synthesis in B16F10 melanoma cells. To elucidate mRNA level reverse transcription polymerase chain reaction (RT-PCR) technique was used. It was revealed that A subfraction of $CHCI_3$ extract of Canavalia lineara reduced the tyrosinase mRNA expression of B16F10 melanoma cells by reverse transcription polymerase chain reaction (RT-PCR) technique.

Anti-inflammatory Effect of Ethanol Extract from Eupatorium japonicum (등골나물 추출물의 항염증 효과)

  • Lee, Han-Na;Lim, Do-Young;Lim, Soon-Sung;Kim, Jong-Dai;Yoon, Jung-Han
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.65-71
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    • 2011
  • Eupatorium japonicum belongs to a family of Asteraceae plants and flowers of E. japonicum have been consumed as a tea. In this study, we investigated whether E. japonicum extract inhibits lipopolysaccharide (LPS)-induced inflammatory responses in Raw264.7 macrophages. The cells were treated with various concentrations (0, 1, 2.5, 5, or 10 mg/L) of 70% ethanol extract from E. japonicum flowers (EJE) in Raw264.7 cells. LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production were inhibited by EJE up to 67% and 49% of these productions, respectively without any reduction of viable cell numbers. EJE reduced LPS-induced expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 proteins and their corresponding mRNA levels. Additionally, EJE decreased the levels of interleukin (IL)-6, IL-1${\beta}$, and tumor necrosis factor (TNF)-${\alpha}$ mRNA. EJE was further fractionated with water, butanol, ethylacetate (EA), hexane, or methylene chloride (MC). Among the resulting five fractions, EA and MC, respectively from EJE significantly inhibited LPS-induced NO production (each inhibition rate was 85.3% of 10 mg/L EA fraction and 97.2% of 10 mg/L MC fraction) without significant cytotoxicity in Raw264.7 cells. These results indicate that EJE exhibits powerful effects of anti-inflammation and can be developed as a potential anti-inflammatory agent.

Anti-oxidant Function and Inhibitory Effects of the Expression of MITF, TRP-1, TRP-2 and Tyrosinase of Sesamum indicum L. in B16F10 Melanoma Cells (참깨 추출물의 항산화 활성 및 melanoma cell (B16F10)에서 MITF, TRP-1, TRP-2, tyrosinase 의 발현 저해)

  • Yoo, Dan-Hee;Joo, Da-Hye;Lee, Jin-Young
    • Journal of Life Science
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    • v.27 no.3
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    • pp.318-324
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    • 2017
  • This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.