• 제목/요약/키워드: RT synthesis

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Synthesis of Methylated and Acetylated Derivatives of Meso-dihydroguaretic Acid and Study of Their Inhibitory Activities on LPS Derived Nitric Oxide (NO) Production (메조-디하이드로구아레틱산 메틸, 아세틸 치환체의 합성 및 이들 화합물들의 LPS에 의해서 유도된 일산화질소(NO)의 억제 효능에 대한 연구)

  • Choi, Kyungoh;Rho, Ho Sik
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.3
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    • pp.195-200
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    • 2017
  • This study was conducted to examine the inhibitory effects of meso-dihydroguaretic acid (MDGA, 1) and its synthetic derivatives (compound 2 and 3) against NO production. MDGA is a lignan component isolated from the bark of Machilus thunbergii Sieb. et Zucc. We synthesized dimethylated MDGA (2), diacetylated MDGA (3) and compared NO inhibition of two derivatives with that of MDGA (1). MDGA (1) and compound 3 showed suppressive effects against the generation of NO in LPS-activated macrophages. RT-PCR analysis suggested that MDGA (1) and compound 3 inhibited NO production through the suppression of iNOS mRNA expression. From these results, diacetylated MDGA (3) can be used as a pro-drug for MDGA.

Inhibitory Effect of Galangin from Alpinia officinarum on Lipopolysaccharide-induced Nitric Oxide Synthesis in RAW 264.7 macrophages (고량강으로부터 분리된 galangin의 RAW 264.7 세포주에서 LPS로 유도된 nitric oxide 생성 저해활성)

  • Lee, Hwa Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.4
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    • pp.511-515
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    • 2014
  • In a screen for plant-derived inhibitors of nitric oxide (NO) production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophage cells, a flavonol isolated from the chloroform extract of Alpinia officinarum was isolated. The structure of the flavonol was found to be 3,5,7-trihydroxy-2-phenylchromen-4-one (galangin, GLG) by using spectroscopy. GLG exhibited an inhibitory effect ($IC_{50}$ value: $26.8{\mu}M$) on NO production in LPS-stimulated RAW 264.7 murine macrophage cells. Moreover, GLG suppressed expressions of inducible nitric oxide synthase (iNOS) protein and mRNA in a dose-dependent manner.

Identification and Expression Analysis of Genes Induced in Response to Tomato chlorosis virus Infection in Tomato

  • Sahin-Cevik, Mehtap;Sivri, Emine Dogus;Cevik, Bayram
    • The Plant Pathology Journal
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    • v.35 no.3
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    • pp.257-273
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    • 2019
  • Tomato (Solanum lycopersicum) is one of the most widely grown and economically important vegetable crops in the world. Tomato chlorosis virus (ToCV) is one of the recently emerged viruses of tomato distributed worldwide. ToCV-tomato interaction was investigated at the molecular level for determining changes in the expression of tomato genes in response to ToCV infection in this study. A cDNA library enriched with genes induced in response to ToCV infection were constructed and 240 cDNAs were sequenced from this library. The macroarray analysis of 108 cDNAs revealed that the expression of 92 non-redundant tomato genes was induced by 1.5-fold or greater in response to ToCV infection. The majority of ToCV-induced genes identified in this study were associated with a variety of cellular functions including transcription, defense and defense signaling, metabolism, energy, transport facilitation, protein synthesis and fate and cellular biogenesis. Twenty ToCV-induced genes from different functional groups were selected and induction of 19 of these genes in response to ToCV infection was validated by RT-qPCR assay. Finally, the expression of 6 selected genes was analyzed in different stages of ToCV infection from 0 to 45 dpi. While the expression of three of these genes was only induced by ToCV infection, others were induced both by ToCV infection and wounding. The result showed that ToCV induced the basic defense response and activated the defense signaling in tomato plants at different stages of the infection. Functions of these defense related genes and their potential roles in disease development and resistance to ToCV are also discussed.

Melanogenesis Inhibition Effect of Rosa multiflora Extracts in B16 Melanoma Cells (찔레 추출물의 B16 세포 멜라닌 형성 억제)

  • Ha, Se-Eun;Kim, Hyoung-Do;Park, Jong-Kun;Chung, Yeon-Ok;Kim, Hyun-Jong;Park, Nou-Bog
    • Korean Journal of Plant Resources
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    • v.22 no.4
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    • pp.317-322
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    • 2009
  • In the present study, the effect of ethanol extracts of leaf and root of Rosa multiflora on the proliferation of B16 cells, tyrosinase activity and melanin synthesis were investigated. 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide(MTT) assay demonstrated that the cell viability upon treatment with Rosa multiflora extract(0-200 ${\mu}g$/ml) was similar to that of untreated control. Treatment with leaf or root extracts(200 ${\mu}g$/ml) decreased the in vitro tyrosinase activity to about 65% of that in untreated control. Similarly, the intracellular tyrosinase activity of B16 cells decreased in a concentration-dependent manner. Furthermore, the melanin synthesis of B16 cells decreased by the two extracts in a concentration dependent manner. However, the extracts did not change the level of tyrosinase mRNA, as determined by RT-PCR. These results demonstrated that the Rosa multiflora extracts inhibit the tyrosinase dependent melanin biosynthesis, and therefore, are candidates for skin-whitening agents.

Synthesis of 2',4'-Dimethoxyflavone and Its Anti-pigmentation Effect (2',4'-Dimethoxyflavone의 합성과 멜라닌 생성 저해 효과)

  • Lee, Bum-Chun;Sim, Gwan-Sub;Kim, Jin-Hwa;Kim, Ki-Ho;Lee, Hwa-Jeong;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.3 s.58
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    • pp.153-160
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    • 2006
  • We synthesized 2',4'-dimethoxyflavone and investigated the effects on melanogenesis. To determine the effects as a whitening agent, various in uitro tests were performed such as free radical scavenging activity, melanin assay, tyrosinase activity and expression of tyrosinase, TRP-1 and TRP-2 (western blot and RT-PCR) in Bl6 melanoma cells. 2',4'-Dimethoxyflavone showed neither free radical scavenging activities against 1,1- diphenyl -2-picrylhydrazvl (DPPH) radical and inhibition of mushroom tyrosinase activity, 2',4'-dimethoxyflavone significantly inhibited melanin production in B16 melanoma cells. 2',4'-Dimethoxyflavone treatment (48h) suppressed the biosynthesis of melanin up to 27% at $5{\mu}g/mL$ and reduced tyrosinase activity up to 20% at $5{\mu}g/mL$ in B16 melanoma cells. 2',4'-Dimethoxyflavone was also able to significantly inhibit tyrosinase and TRP-1 expression in protein and mRNA level. These results suggest that 2',4'-dimethoxyflavone inhibits melanin biosynthesis at the level of enzyme activity and protein mRNA expression B16 melanoma cells. Therefore, 2',4'-dimethoxyflavone may be useful as a new whitening agent in cosmetics.

Sedum sarmentosum Enhances Hyaluronan Synthesis in Transformed Human Keratinocytes and Increases Water Content in Human Skin (돌나물추출물에 의한 사람 각질형성세포에서의 Hyaluronan Synthesis 촉진과 인체 피부의 보습력 증진)

  • Sim, Gwan-Sub;Kim, Jin-Hwa;Lee, Dong-Hwan;Na, Young;Lee, Geun-Soo;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.1 s.60
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    • pp.17-22
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    • 2007
  • In this study, we investigated the effects of Sedum sarmentosum extract on the expression of hyaluronan synthase (HAS) genes and hyaluronan (HA) production in HaCaT keratinocytes. We also assessed water content (electrical capacitance) and transepidermal water loss (TEWL) in human skin after topical treatment with Sedum sarmentosum extract. Sedum sarmentosum extract increased expression of HAS-2 and HAS-3 genes in HaCaT cells, when assayed by real-time reverse-transcriptase polymerase chain reactions (RT-PCR). Sedum sarmentosum extract induced HA production in HaCaT cells, when determined by enzyme-linked immunosorbent assay (ELISA). Finally, treatment of Sedum sarmentosum extract on human skin increased the skin hydration and decreased TEWL when measured using Corneometer and Tewameter. Our study suggests that Sedum sarmentosum extract should be a very useful cosmetic ingredient, as a skin moisturizer.

miR-380-3p promotes β-casein expression by targeting αS1-casein in goat mammary epithelial cells

  • Ning Song;Jun Luo;Lian Huang;Xiaoying Chen;Huimin Niu;Lu Zhu
    • Animal Bioscience
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    • v.36 no.10
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    • pp.1488-1498
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    • 2023
  • Objective: αS1-Casein is more closely associated with milk allergic reaction than other milk protein components. microRNA (miRNA) is a class of small non-coding RNAs that modulate multiple biological progresses by the target gene. However, the post-transcriptional regulation of αS1-casein expression by miRNA in ruminants remains unclear. This study aims to explore the regulatory roles of miR-380-3p on αS1-casein synthesis in goat mammary epithelial cells (GMEC). Methods: αS1-Casein gene and miR-380-3p expression was measured in dairy goat mammary gland by quantitative real-time polymerase chain reaction (qRT-PCR). miR-380-3p overexpression and knockdown were performed by miR-380-3p mimic or inhibitor in GMEC. The effect of miR-380-3p on αS1-casein synthesis was detected by qRT-PCR, western blot, luciferase and chromatin immunoprecipitation assays in GMEC. Results: Compared with middle-lactation period, αS1-casein gene expression is increased, while miR-380-3p expression is decreased during peak-lactation of dairy goats. miR-380-3p reduces αS1-casein abundance by targeting the 3'-untranslated region (3'UTR) of αS1-casein mRNA in GMEC. miR-380-3p enhances β-casein expression and signal transducer and activator of transcription 5a (STAT5a) activity. Moreover, miR-380-3p promotes β-casein abundance through target gene αS1-casein, and activates β-casein transcription by enhancing the binding of STAT5 to β-casein gene promoter region. Conclusion: miR-380-3p decreases αS1-casein expression and increases β-casein expression by targeting αS1-casein in GMEC, which supplies a novel strategy for reducing milk allergic potential and building up milk quality in ruminants.

Effect of bFGF and fibroblasts combined with hyaluronic acid-based hydrogels on soft tissue augmentation: an experimental study in rats

  • Lee, Su Yeon;Park, Yongdoo;Hwang, Soon Jung
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.41
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    • pp.47.1-47.10
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    • 2019
  • Background: Hyaluronic acid (HA) has been applied as a primary biomaterial for temporary soft tissue augmentation and as a carrier for cells and the delivery of growth factors to promote tissue regeneration. Although HA derivatives are the most versatile soft tissue fillers on the market, they are resorbed early, within 3 to 12 months. To overcome their short duration, they can be combined with cells or growth factors. The purpose of this study was to investigate the stimulating effects of human fibroblasts and basic fibroblast growth factors (bFGF) on collagen synthesis during soft tissue augmentation by HA hydrogels and to compare these with the effects of a commercial HA derivative (Restylane®). Methods: The hydrogel group included four conditions. The first condition consisted of hydrogel (H) alone as a negative control, and the other three conditions were bFGF-containing hydrogel (HB), human fibroblast-containing hydrogel (HF), and human fibroblast/bFGF-containing hydrogel (HBF). In the Restylane® group (HGF), the hydrogel was replaced with Restylane® (R, RB, RF, RBF). The gels were implanted subdermally into the back of each nude mouse at four separate sites. Twelve nude mice were used for the hydrogel (n = 6) and Restylane® groups (n = 6). The specimens were harvested 8 weeks after implantation and assessed histomorphometrically, and collagen synthesis was evaluated by RT-PCR. Results: The hydrogel group showed good biocompatibility with the surrounding tissues and stimulated the formation of a fibrous matrix. HBF and HF showed significantly higher soft tissue synthesis compared to H (p < 0.05), and human collagen type I was well expressed in HB, HF, and HBF; HBF showed the strongest expression. The Restylane® filler was surrounded by a fibrous capsule without any soft tissue infiltration from the neighboring tissue, and collagen synthesis within the Restylane® filler could not be observed, even though no inflammatory reactions were observed. Conclusion: This study revealed that HA-based hydrogel alone or hydrogel combined with fibroblasts and/or bFGF can be effectively used for soft tissue augmentation.

Alfalfa xenomiR-162 targets G protein subunit gamma 11 to regulate milk protein synthesis in bovine mammary epithelial cells

  • Guizhi Meng;Hongjuan Duan;Jingying Jia;Baobao Liu;Yun Ma;Xiaoyan Cai
    • Animal Bioscience
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    • v.37 no.3
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    • pp.509-521
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    • 2024
  • Objective: It was shown that microRNAs (miRNAs) play an important role in milk protein synthesis. However, the post-transcriptional regulation of casein expression by exogenous miRNA (xeno-miRNAs) in ruminants remains unclear. This study explores the regulatory roles of alfalfa xeno-miR162 on casein synthesis in bovine mammary epithelial cells (bMECs). Methods: The effects of alfalfa xenomiR-162 and G protein subunit gamma 11 (GNG11) on proliferation and milk protein metabolism of bMECs were detected by 5-Ethynyl-2'-Deoxyuridine (EdU) staining, flow cytometry, cell counting kit-8 (CCK-8), enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot. Dual-luciferase reporter assay was used to verify the targeting relationship between GNG11 and xenomiR-162. Results: Results showed that over-expression of xenomiR-162 inhibited cell proliferation but promoted apoptosis, which also up-regulated the expression of several casein coding genes, including CSN1S1, CSN1S2, and CSN3, while decreasing the expression of CSN2. Furthermore, the targeting relationship between GNG11 and xenomiR-162 was determined, and it was confirmed that GNG11 silencing also inhibited cell proliferation but promoted apoptosis and reduced the expression of casein coding genes and genes related to the mammalian target of rapamycin (mTOR) pathway. Conclusion: Alfalfa xenomiR-162 appears to regulate bMECs proliferation and milk protein synthesis via GNG11 in the mTOR pathway, suggesting that this xeno-miRNA could be harnessed to modulate CSN3 expression in dairy cows, and increase κ-casein contents in milk.

Characterization of Embryo-specific Autophagy during Preimplantation (착상전 난자 자식작용의 특성규명)

  • Lee, Jae-Dal
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.12 no.8
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    • pp.3541-3546
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    • 2011
  • Autophagy is an evolutionarily conserved lysosomal pathway for degrading cytoplasmic proteins, macromolecules, and organelles in addition to recycling protein and ATP synthesis. Although autophagy is very important during embryogenesis, the mechanism underlying the dynamic development during this process remains largely unknown. In order to obtain insights into autophagy in early embryo development, we analyzed gene expression levels of autophagy-related genes (ATGs) in mouse embryos developing in vitro. Using real time RT-PCR technique, ATGs including Atg2a, Atg3, Atg4b, Atg5, Atg6, Atg7, Atg9a, and Wipi3, as maternal transcripts, were only up-regulated in 1-cell embryo stage before zygotic genomic activation (ZGA), and then expression decreased from 2-cell to blastocyst embryo stage. ATGs including Dram and Atg9b were expressed abundantly in 1-cell embryo state and in blastocyst embryo stage, athough Atg8 and Ulk1 were constantly expressed during preimplantation stage. However, Atg4d were only up-expressed from 4-cell to blastocyst stage. These results suggest that autophagy is related in mouse embryo, which possibly gives an important role for early development.