• Title/Summary/Keyword: mRNA levels

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Improvement of colitis preventive effects of Gochujang by addition of Lactobacillus plantarum on C57BL/6 mice (Lactobacillus plantarum 첨가 고추장의 C57BL/6 마우스에서 대장염 예방 증진효과)

  • Park, Eui-Seong;Heo, Ju-Hee;Lim, Yaung-Iee;Ju, Jaehyun;Park, Kun-Young
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1188-1194
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    • 2017
  • Gochujang, a traditional Korean food, is fermented by mixing red pepper powder, various grain, meju and salt. Changes in the kind of ingredients and fermentation method could increase health functionalities. In this study, in vivo anti-colitis effects of gochujang prepared with mixed grains, bamboo salt baked 3 times and meju starters on DSS-induced colitis in C57BL/6 mice were studied. We prepared gochujang prepared with mixed grains (MG), bamboo salt, and Aspergillus oryzae (A) and Baccillus subtilis (B) mixed starters (MG-AB) and gochujang prepared with MG, bamboo salt and A, B and Lactobacillus plantarum (L) mixed starters (MG-ABL). MG-AB and MG-ABL significantly increased body weight and colon length compared to the control (p<0.05). MG-ABL showed significantly decreased interleukin-6 (IL-6) expression in serum compared to the control and MG-AB group (p<0.05). MG-ABL also regulated mRNA and protein levels of pro-apoptotic Bcl-2-associated X protein (Bax) and anti-apoptotic B-cell lymphoma-2 (Bcl-2) in the mice colon tissue (p<0.05). Therefore, MG-ABL exhibited the increased anticolitis effects by inhibiting damage of colon tissue, probably by regulating a pro-inflammatory cytokine of IL-6 and regulated apoptosis related genes. These results indicated that gochujang changed with good ingredients and starters had colitis preventive effects and might be due to active compounds in mixed grain and bamboo salt, and produced by L during the fermentation of gochujang.

In vitro antioxidative and anti-inflammatory effects of the compound K-rich fraction BIOGF1K, prepared from Panax ginseng

  • Hossen, Muhammad Jahangir;Hong, Yong Deog;Baek, Kwang-Soo;Yoo, Sulgi;Hong, Yo Han;Kim, Ji Hye;Lee, Jeong-Oog;Kim, Donghyun;Park, Junseong;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.43-51
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    • 2017
  • Background: BIOGF1K, a compound K-rich fraction prepared from the root of Panax ginseng, is widely used for cosmetic purposes in Korea. We investigated the functional mechanisms of the anti-inflammatory and antioxidative activities of BIOGF1K by discovering target enzymes through various molecular studies. Methods: We explored the inhibitory mechanisms of BIOGF1K using lipopolysaccharide-mediated inflammatory responses, reporter gene assays involving overexpression of toll-like receptor adaptor molecules, and immunoblotting analysis. We used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay to measure the antioxidative activity. We cotransfected adaptor molecules, including the myeloid differentiation primary response gene 88 (MyD88) and Toll/interleukin-receptor domain containing adaptor molecule-inducing interferon-${\beta}$ (TRIF), to measure the activation of nuclear factor (NF)-${\kappa}B$ and interferon regulatory factor 3 (IRF3). Results: BIOGF1K suppressed lipopolysaccharide-triggered NO release in macrophages as well as DPPH-induced electron-donating activity. It also blocked lipopolysaccharide-induced mRNA levels of interferon-${\beta}$ and inducible nitric oxide synthase. Moreover, BIOGF1K diminished the translocation and activation of IRF3 and NF-${\kappa}B$ (p50 and p65). This extract inhibited the upregulation of NF-${\kappa}B$-linked luciferase activity provoked by phorbal-12-myristate-13 acetate as well as MyD88, TRIF, and inhibitor of ${\kappa}B$ ($I{\kappa}B{\alpha}$) kinase ($IKK{\beta}$), and IRF3-mediated luciferase activity induced by TRIF and TANK-binding kinase 1 (TBK1). Finally, BIOGF1K downregulated the NF-${\kappa}B$ pathway by blocking $IKK{\beta}$ and the IRF3 pathway by inhibiting TBK1, according to reporter gene assays, immunoblotting analysis, and an AKT/$IKK{\beta}$/TBK1 overexpression strategy. Conclusion: Overall, our data suggest that the suppression of $IKK{\beta}$ and TBK1, which mediate transcriptional regulation of NF-${\kappa}B$ and IRF3, respectively, may contribute to the broad-spectrum inhibitory activity of BIOGF1K.

The Effects of Somatid on the Cytotoxicity of Cancer Cells and Human Papillomavirus Type 16 E6 and E7 Oncogenes (생기액(生肌液)의 세포독성 및 자궁경부암 바이러스 (HPV 16 type) 암 유발인자 E6와 E7의 작용에 미치는 효과)

  • Joung, Ok;Cho, Young-Sik;Cho, Cheong-Weon;Lee, Kyung-Ae;Shim, Jung-Hyun;Cho, Min-Chul;Lee, Hong-Soo;Yeom, Young-Il;Kim, Sang-Bom;Park, Sue-Nie;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.44 no.4
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    • pp.340-346
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    • 2000
  • Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, named as Somatid, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Somatid inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, CaSki) and HaCaT keratinocytes in a dose response manner, In vitro binding assay and ELISA showed that Somatid inhibited the in vitro biding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Somatid inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by Somatid. Our data suggested that Somatid inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas caused by HPV.

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Effects of Unripe Black Raspberry Water Extract on Lipid Metabolism and Oxidative Stress in Mice (복분자 미숙과 물 추출물이 마우스의 지질대사 및 산화적 스트레스에 미치는 영향)

  • Choi, Hye Ran;Lee, Jung-Hyun;Lee, Su Jung;Lee, Min Jung;Jeong, Jong Tae;Lee, Tae-Bum
    • Korean Journal of Food Science and Technology
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    • v.46 no.4
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    • pp.489-497
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    • 2014
  • We examined the effects of unripe black raspberry water extract (UBR-W) on lipid metabolism and oxidative stress in mice. C57BL/6J mice were divided into 4 groups: those administered a control diet (CTL), high-fat diet (HFD), UBR-W and simvastatin for 12 weeks. In the HFD group, LDL cholesterol were significantly higher than in the CTL group. However, the UBR-W treated group showed dose-dependent reduction of plasma LDL levels. Hepatic total lipid, TC, and malondialdehyde were significantly increased in hyperlipidemic mice. However, supplementation with either UBR-W or simvastatin effectively reduced these lipid profiles and lipid peroxidation. UBR-W increased mRNA expression of the LDL receptor, sterol regulatory element binding protein 2 (SREBP2), 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase and ATP-binding cassette transporter A1 (ABCA1) compared to that observed in the HFD group. In addition, UBR-W and simvastatin showed significantly reduced oxidized LDL uptake by the scavenger receptor CD36. These results suggest that UBR-W is useful for treatment and prevention of hyperlipidemia and lipid peroxidation.

17β-estradiol mediated effects on pluripotency transcription factors and differentiation capacity in mesenchymal stem cells derived porcine from newborns as steroid hormones non-functional donors

  • Lee, Won-Jae;Park, Ji-Sung;Lee, HyeonJeong;Lee, Seung-Chan;Lee, Jeong-Hyun;Ock, Sun-A;Rho, Gyu-Jin;Lee, Sung-Lim
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.209-220
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    • 2017
  • The estrogen-mediated effect of mesenchymal stem cells (MSCs) is a highly critical factor for the clinical application of MSCs. However, the present study is conducted on MSCs derived from adult donors, which have different physiological status with steroid hormonal changes. Therefore, we explores the important role of $17{\beta}$-estradiol (E2) in MSCs derived from female and male newborn piglets (NF- and NM-pBMSCs), which are non-sexually matured donors with steroid hormones. The results revealed that in vitro treatment of MSCs with E2 improved cell proliferation, but the rates varied according to the gender of the newborn donors. Following in vitro treatment of newborn MSCs with E2, mRNA levels of Oct3/4 and Sox2 increased in both genders of MSCs and they may be correlated with both estrogen receptor ${\alpha}$ ($ER{\alpha}$) and $ER{\beta}$ in NF-pBMSCs, but NM-pBMSCs were only correlated with $ER{\alpha}$. Moreover, E2-treated NF-pBMSCs decreased in ${\beta}$-galactosidase activity but no influence on NM-pBMSCs. In E2-mediated differentiation capacity, E2 induced an increase in the osteogenic and chondrogenic abilities of both pBMSCs, but adipogenic ability may increased only in NF-pBMSCs. These results demonstrate that E2 could affect both genders of newborn donor-derived MSCs, but the regulatory role of E2 varies depending on gender-dependent characteristics even though the original newborn donors had not been affected by functional steroid hormones.

Protective Effects of the Ethanol Extract of Viola tianshanica Maxim against Acute Lung Injury Induced by Lipopolysaccharides in Mice

  • Wang, Xue;Yang, Qiao-Li;Shi, Yu-Zhu;Hou, Bi-Yu;Yang, Sheng-Qian;Huang, Hua;Zhang, Li;Du, Guan-Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1628-1638
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    • 2017
  • Viola tianshanica Maxim, belonging to the Violaceae plant family, is traditionally used in Uighur medicine for treating pneumonia, headache, and fever. There is, however, a lack of basic understanding of its pharmacological activities. This study was designed to observe the effects of the ethanol extract (TSM) from Viola tianshanica Maxim on the inflammation response in acute lung injury (ALI) induced by LPS and the possible underlying mechanisms. We found that TSM (200 and 500 mg/kg) significantly decreased inflammatory cytokine production and the number of inflammatory cells, including macrophages and neutrophils, in bronchoalveolar lavage fluid. TSM also markedly inhibited the lung wet-to-dry ratio and alleviated pathological changes in lung tissues. In vitro, after TSM ($12.5-100{\mu}g/ml$) treatment to RAW 264.7 cells for 1 h, LPS ($1{\mu}g/ml$) was added and the cells were further incubated for 24 h. TSM dose-dependently inhibited the levels of proinflammatory cytokines, such as NO, $PGE_2$, $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$, and remarkably decreased the protein and mRNA expression of $TNF-{\alpha}$ and IL-6 in LPS-stimulated RAW 264.7 cells. TSM also suppressed protein expression of $p-I{\kappa}Ba$ and p-ERK1/2 and blocked nuclear translocation of $NF-{\kappa}B$ p65. The results indicate that TSM exerts anti-inflammatory effects related with inhibition on $NF-{\kappa}B$ and MAPK (p-ERK1/2) signaling pathways. In conclusion, our data demonstrate that TSM might be a potential agent for the treatment of ALI.

Identification of Brassinosteroid-Related Protein, BAK1 from Nutrition Deficient Tomato Cultivated by Soilless Cultivation System (수경재배 영양결핍토마토에서 브레시노스테로이드관련 신호전달 단백질 BAK1의 동정)

  • Shin, Pyung-Gyun;Chang, An-Cheol;Hong, Sung-Chang;Lee, Ki-Sang
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1729-1733
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    • 2007
  • Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.

Molecular Cloning and Expression of a Cu/Zn-Containing Superoxide Dismutase from Thellungiella halophila

  • Xu, Xiaojing;Zhou, Yijun;Wei, Shanjun;Ren, Dongtao;Yang, Min;Bu, Huahu;Kang, Mingming;Wang, Junli;Feng, Jinchao
    • Molecules and Cells
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    • v.27 no.4
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    • pp.423-428
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    • 2009
  • Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

Anti-allergic Effect of Eckolona cava Ethyl Acetate Fraction of on IgE/BSA-stimulated Bone Marrow-derived Cultured Mast Cells (IgE/BSA가 자극한 골수유래 비만 세포에 대한 감태 Ethyl Acetate 분획물의 항알러지 효능)

  • Han, Eui Jeong;Kim, Hyun Soo;Shin, Eun Ji;Kim, Min Ju;Han, Hee-Jin;Jeon, You-Jin;Jee, Youngheun;Ahn, Ginnae
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.277-284
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    • 2018
  • In this study, we investigated the anti-allergic effect of the ethyl acetate fraction of Ecklonia cava (EC-EtoAc) on the immunoglobulin E (IgE)/bovine serum albumin (BSA)-mediated activation of bone marrow-derived cultured mast cells (BMCMCs). We revealed that the $62.5{\mu}g/ml$ of EC-fractions ($EC-CHCl_3$, EC-Hexane and EC-EtoAc) inhibited IgE/BSA-activated ${\beta}$-hexosaminidase release from BMCMCs without cytotoxicity. Especially, EC-EtoAc showed the higher ${\beta}$-hexosaminidase release than the others. Also, EC-EtoAc reduced the expression levels of cytokines such as interleukin (IL)-$1{\beta}$, IL-4, IL-5, IL-6, IL-10, IL-13, interferon (IFN)-${\gamma}$ and tumor necrosis factor (TNF)-${\alpha}$ and a chemokine, thymus- and activation-regulated chemokine (TARC), compared to the only IgE/BSA-treated BMCMCs. Furthermore, EC-EtoAc significantly prevented the binding of IgE to Fc epsilon receptor $(Fc{\varepsilon}R)I$ and reduced the $Fc{\varepsilon}RI$ expression on the sensitized BMCMCs. Taken together, these results suggest that E. cava may be the natural agent with beneficial potentials for the treatment of type I allergic diseases induced by mast cell activation.

Angiopoietin-1 and -2 and vascular endothelial growth factor expression in ovarian grafts after cryopreservation using two methods

  • Cho, In Ae;Lee, Yeon Jee;Lee, Hee Jung;Choi, In Young;Shin, Jeong Kyu;Lee, Soon Ae;Lee, Jong Hak;Choi, Won Jun
    • Clinical and Experimental Reproductive Medicine
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    • v.45 no.3
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    • pp.143-148
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    • 2018
  • Objective: The favored method of preserving fertility in young female cancer survivors is cryopreservation and autotransplantation of ovarian tissue. Reducing hypoxia until angiogenesis takes place is essential for the survival of transplanted ovarian tissue. The aim of this study was to investigate the role of angiopoietin-1 (Angpt-1), angiopoietin-2 (Angpt-2), and vascular endothelial growth factor (VEGF) in ovarian tissue grafts that were cryopreserved using two methods. Methods: Ovarian tissues harvested from ICR mice were divided into three groups: group I (control), no cryopreservation; group II, vitrification in EFS (ethylene-glycol, ficoll, and sucrose solution)-40; and group III, slow freezing in dimethyl sulfoxide. We extracted mRNA for VEGF, Angpt-1, and Angpt-2 from ovarian tissue 1 week following cryopreservation and again 2 weeks after autotransplantation. We used reverse transcriptase-polymerase chain reaction to quantify the levels of VEGF, Angpt-1, and Angpt-2 in the tissue. Results: Angpt-1 and Angpt-2 expression decreased after cryopreservation in groups II and III. After autotransplantation, Angpt-1 and Angpt-2 expression in ovarian tissue showed different trends. Angpt-1 expression in groups II and III was lower than in group I, but Angpt-2 in groups II and III showed no significant difference from group I. The vitrified ovarian tissues had higher expression of VEGF and Angpt-2 than the slow-frozen ovarian tissues, but the difference was not statistically significant. Conclusion: Our results indicate that Angpt-2 may play an important role in ovarian tissue transplantation after cryopreservation although further studies are needed to understand its exact function.