• Title/Summary/Keyword: MTS Assay

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Anti-proliferative Activities of Solvent Fractions of Lees Extracts in Human Colorectal HCT116 Cells (대장암 세포주에서 주박 추출물의 유기용매 분획물의 항성장 활성)

  • Kang, Hyung-Taek;Lee, Seung Hoon;Kim, Soon Young;Kim, Mi-Sun;Shin, Woo-Chang;Sohn, Ho-Yong;Kim, Jong-Sik
    • Journal of Life Science
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    • v.24 no.9
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    • pp.967-972
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    • 2014
  • In the present study, we prepared eighty-five different kinds of lees extracts and their solvent fractions and investigated their anti-proliferative activities against human colorectal cancer HCT116 cells. HCT116 cells were treated with eighty-five solvent fractions of lees extracts and then cell viability was measured using MTS assay. Among the treated solvent fractions, three solvent fractions (KSD-E1-3, KSD-E2-3, and KSD-E4-3) were selected based on cell viability assay. In addition, we performed an oligo DNA microarray analysis to analyze the gene expression changes by treatment of KSD-E1-3 in HCT116 cells. Among the upregulated genes, we selected 4 genes (NAG-1, ATF3, p21, and DDIT3) and performed RT-PCR using gene-specific primers. Among the treated solvent fractions, KSD-E1-3 dramatically induced the expressions of the four selected genes. In addition, we investigated whether the upregulations of those genes were dependent on the transcription factor p53's presence using p53 null HCT116 cells. The results indicate that the upregulations of NAG-1, ATF3, and DDIT3 are not dependent on the p53 presence, whereas p21 is dependent on the p53 presence. These findings may help to understand the molecular mechanisms of the anti-proliferative activity mediated by rice wine lees in human colorectal cancer cells.

The Study of Anti-inflammatory Effect of Suryeon-hwan Water Extract in RAW 264.7 Cells (대식세포에서 수련환(茱連丸) 물추출물의 항염증작용에 관한 연구)

  • Yoon, Yeo-Hwan;Kim, Sung-Bae;Kang, Ok-Hwa;Mun, Su-Hyun;Seo, Yun-Soo;Yang, Da-Wun;Kang, Da-Hye;Wi, Gyeong;Lim, Jae-Soo;Kim, Ma-Ryong;Kwak, Nam-Won;Kong, Ryong;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.29 no.6
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    • pp.125-132
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    • 2014
  • Objectives : Suryeon-hwan (SRH) exhibits potent anti-inflammatory activity with an unknown mechanism. However, there has been a lack of studies regarding the effects of SRH on the inflammatory activities and effector inflammatory disease mechanism about macrophage before is not known. So, the investigation focused on whether SRH inhibited nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) productions, as well as the expressions of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with 200 ng/mL of LPS 30 min prior to the addition of SRH. Cell viability was measured by MTS assay. The production of nitric oxide (NO) was determined by reacting cultured medium with Griess reagent. The content of level of cytokines (PGE, IL-6) in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. Results : We found that SRH inhibited LPS-induced NO, $PGE_2$ and IL-6 productions as well as the expressions of iNOS and COX-2. Furthermore, SRH suppressed the LPS-induced phosphorylation of MAPK and extracellular signal-regulated kinase 1/2 (ERK 1/2) activation. Conclusions : These results suggest that SRH has inhibitory effects on LPS-induced $PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the phosphorylation of MAPKs following activation.

Pre-treatment of the White-Spotted Flower Chafer (Protaetia brevitarsis) as an Ingredient for Novel Foods (흰점박이꽃무지(Protaetia brevitarsis)의 식품원료화를 위한 전처리 조건 확립)

  • Kwon, Eun-Young;Yoo, Jeongmi;Yoon, Young-Il;Hwang, Jae-Sam;Goo, Tae-Won;Kim, Mi-Ae;Choi, Young-Cheol;Yun, Eun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.397-402
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    • 2013
  • The pharmacological efficacy of Protaetia (P.) brevitarsis larvae has been described in the Dongui Bogam. It is believed that the larvae are particularly useful for hepatic disorders. However, natural aversion has made it difficult to consume these larvae as food. Thus, we sought to make an eatable form of the larvae by establishing optimal conditions for larvae preparation. Larvae were selectively bred, sterilized, and a powder of larvae generated by freeze-drying. Afterward, the CellTiter $96^{(R)}$ AQueous Non-Radioactive Cell Proliferation Assay (MTS) with the RAW 264.7 cell line was used to validate the safety of the powder as a food ingredient. We determined that oak sawdust sterilized by water vapor for 5 minutes could be used for larvae feed, and a feeding for 3~5 days followed by a fasting for 3 days were optimal conditions for larvae preparation. In addition, sterilization of larvae at $115^{\circ}C$ and $0.9kgf/cm^3$ (to avoid contamination of pathogenic bacteria and fungi) was successfully applied in the production of edible powder from P. brevitarsis. The optimized processes established in our experiments can be used in the industrial production of P. brevitarsis as a food ingredient.