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Inhibitory Effect of Ethanol Extract of Monascus-fermented Red Yeast Rice on Proinflammatory iNOS and COX-2 Protein Expression in LPS-stimulated RAW 264.7 Macrophage Cells (Monascus sp. BHN-MK로 발효생산한 홍국 에탄올 추출물의 Raw 264.7 대식세포에 있어 친-염증성 iNOS와 COX-2 단백질 발현 억제 효과)

  • Kim, Ki Hyun;Lee, Jung-Hyeong;Kwon, Gi-Seok;Seo, Eul Won;Lee, Jung-Bok
    • Journal of Life Science
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    • v.30 no.4
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    • pp.352-358
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    • 2020
  • Red yeast rice has been extensively used as a food and traditional medicine for thousands of years in Korea. Monascus produces many secondary metabolites during its growth, including pigments, monacolins, and γ-aminobutyric acid. Some metabolites, specifically monacolin K, γ-aminobutyric acid, and dimerumic acid, have been reported to lower cholesterol and blood pressure because of certain antioxidant effects. This study investigated the total phenolic content of ethanol extract from red yeast rice fermented with Monascus sp. BHN-MK and its anti-inflammatory effect on LPS-stimulated RAW 264.7 macrophage cells. To assess its anti-inflammatory effect, the inhibitory activity of the ethanol extract on LPS-induced NO production and expression levels of iNOS and COX-2 proteins in macrophage cells were measured. Its total polyphenol content was higher than that of ordinary non-fermented rice. Its NO production inhibition activity was comparable to that of the negative control group treated with LPS at a concentration of 400 ㎍/ml. Western blot revealed a significant decrease in the inhibition of iNOS and COX-2 protein expression at concentrations of 400 and 800 ㎍/ml, respectively. Red yeast rice ethanol extracts exerted the strongest anti-inflammatory effects. The results indicate that red yeast rice could be used as a functional cosmetic and anti-inflammatory material.

MiR-29a and MiR-140 Protect Chondrocytes against the Anti-Proliferation and Cell Matrix Signaling Changes by IL-1β

  • Li, Xianghui;Zhen, Zhilei;Tang, Guodong;Zheng, Chong;Yang, Guofu
    • Molecules and Cells
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    • v.39 no.2
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    • pp.103-110
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    • 2016
  • As a degenerative joint disease, osteoarthritis (OA) constitutes a major cause of disability that seriously affects the quality of life of a large population of people worldwide. However, effective treatment that can successfully reverse OA progression is lacking until now. The present study aimed to determine whether two small non-coding RNAs miR-29a and miR-140, which are significantly down-regulated in OA, can be applied together as potential therapeutic targets for OA treatment. MiRNA synergy score was used to screen the miRNA pairs that potentially synergistically regulate OA. An in vitro model of OA was established by treating murine chondrocytes with IL-$1{\beta}$. Transfection of miR-29a and miR-140 via plasmids was investigated on chondrocyte proliferation and expression of nine genes such as ADAMTS4, ADAMTS5, ACAN, COL2A1, COL10A1, MMP1, MMP3, MMP13 and TIMP metallopeptidase inhibitor 1 (TIMP1). Western blotting was used to determine the protein expression level of MMP13 and TIMP1, and ELISA was used to detect the content of type II collagen. Combined use of miR-29a and miR-140 successfully reversed the destructive effect of IL-$1{\beta}$ on chondrocyte proliferation, and notably affected the MMP13 and TIMP1 gene expression that regulates extracellular matrix. Although co-transfection of miR-29a and miR-140 did not show a synergistic effect on MMP13 protein expression and type II collagen release, but both of them can significantly suppress the protein abundance of MMP13 and restore the type II collagen release in IL-$1{\beta}$ treated chondrocytes. Compared with single miRNA transfection, cotransfection of both miRNAs exceedingly abrogated the suppressed the protein production of TIMP1 caused by IL-$1{\beta}$, thereby suggesting potent synergistic action. These results provided1novel insights into the important function of miRNAs' collaboration in OA pathological development. The reduced MMP13, and enhanced TIMP1 protein production and type II collagen release also implies that miR-29a and miR-140 combination treatment may be a possible treatment for OA.

Study on maintaining healthy body and changes of human body by circadian rhythm (인체(人體)의 일주리듬에 따른 변화(變化)와 건강법(健康法)에 관한 연구(硏究))

  • Jeong, Sang Jee;Kang, Jung Soo
    • Journal of Haehwa Medicine
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    • v.12 no.1
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    • pp.103-121
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    • 2003
  • Human being can't live without nature, then the changes of nature affect human body. It means that human body has corresponding changes to the KI(vital energy) of nature. There is a stream of changes in human body which circulate mysteriously and punctually by the laws of nature. If this stream of changes fits into human's life style, it would be most effective. It has a certain mode continuously. So if a person has a habit fitting into it, he will get the healthiest body. Then the researcher tries to explain the changes in human body by the time, mainly focused on within 24 hours. it is showing not only the oriental view, but also the western's. The researcher can find the coincidence as followings. At In-Si(3-5 am), the body function and the body temperature get to the bottom, therefore it's good for him to wake up and to run the vital energy. At Sa-Si(9-11 am), the patience on pain anxiety and the psychic concentration get to the top, he'd better start the work. At O-Si(11am-1pm), the heart energy has a vital move, then the blood concentration of Hb(hemoglobin) gets to the top. At Mi-Si(1-3 pm), the muscle strength, the squeeze, and the breathing rate increase. The reflex nerve sensitivity gets to the top. Creativity, observation, and working efficiency go high, so it's time to work hard. At Hae-Si(9pm-1am), the body function falls, sleeping is needed. At Chuck-Si(1-3 am), the cell spontaneity gets to the top, immune lymphocyte moves actively, and the blood concentration of growth hormone gets to the top. These are liver's work. In west, there has been active studies on how to reduce the side effect by using a person's bio-rhythm based on the 'time treatment', and how to reorganize the bio-rhythm by using the machine and the age resistance based on the 'bio-watch'. Though the 'time treatment' means something, the artificial resistance on bio-rhythm seems to give bad effects to human body. If a person lives by regimen of oriental medicine, he will maintain the healthiest body. Regimen is that human body follows the laws of nature, and moves its mysterious, Punctual and periodical changes.

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Antiinflammatory Effects of New Chemical Compounds, HS-1580 Series (HS-1580, HS-1581, HS-1582) (신화학물질 HS-1580 유도체(HS-1580 HS-1581 HS-1582)의 항염증 효과)

  • Kim, Ji-Young;Kim, Kyun-Ha;Suh, Hong-Suk;Choi, Won-Chul
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1181-1187
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    • 2006
  • HS-1580 series (HS-1580, HS-1581, HS-1582) can produce anti-imflammatory effects were synthesized from the marine algae extraction in 2,3,6-tribromo-4,5-dihydroxy benzyl methyl ether (TDB). Raw 264.7 cells were pre-treated with $1{\mu}g/{\mu}l$ lipopolysaccharide (LPS) and later treated with HS-1580 series. These cells of inflammatory mediators were tested as well. Nitric oxide (NO) is related to autoimmune disease and is produced by inducible NOS (iNOS). When treated with HS-1580 series, the product of NO will reduce in a dose-dependent manner. HS-1580 series significantly inhibit the iNOS protein expression. Cyclooxygenase (COX) involves with the various physiologic events and catalyzes in prostaglandin. HS-1580 series also inhibit the COX-2 protein expression as well as pro-inflammatory cytokines production such as tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})\;and\;interluekin-1{\alpha]\;(IL-1{\beta})$. These upcoming results suggest that HS-1580 series have anti-inflammatory efforts in Raw 264.7 cells by inhibiting such as iNOS, COX-2, $TNF-{\alpha}\;and\;IL-1{\beta}$ as inflammatory mediators.

Whitening and inhibiting NF-κB-mediated inflammation properties of the biotransformed green ginseng berry of new cultivar K1, ginsenoside Rg2 enriched, on B16 and LPS-stimulated RAW 264.7 cells

  • Xu, Xing Yue;Yi, Eun Seob;Kang, Chang Ho;Liu, Ying;Lee, Yeong-Geun;Choi, Han Sol;Jang, Hyun Bin;Huo, Yue;Baek, Nam-In;Yang, Deok Chun;Kim, Yeon-Ju
    • Journal of Ginseng Research
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    • v.45 no.6
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    • pp.631-641
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    • 2021
  • Background: Main bioactive constituents and pharmacological functions of ripened red ginseng berry (Panax ginseng Meyer) have been frequently reported. Yet, the research gap targeting the beneficial activities of transformed green ginseng berries has not reported elsewhere. Methods: Ginsenosides of new green berry cultivar K-1 (GK-1) were identified by HPLC-QTOF/MS. Ginsenosides bioconversion in GK-1 by bgp1 enzyme was confirmed with HPLC and TLC. Then, mechanisms of GK-1 and β-glucosidase (bgp1) biotransformed GK-1 (BGK-1) were determined by Quantitative Reverse Transcription-Polymerase Chain Reaction and Western blot. Results: GK-1 possesses highest ginsenosides especially ginsenoside-Re amongst seven ginseng cultivars including (Chunpoong, Huangsuk, Kumpoong, K-1, Honkaejong, Gopoong, and Yunpoong). Ginseng root's biomass is not affected with the harvest of GK-1 at 3 weeks after flowering period. Then, Re is bioconverted into a promising pharmaceutical effect of Rg2 via bgp1. According to the results of cell assays, BGK-1 shows decrease of tyrosinase and melanin content in α-melanocyte-stimulating hormone challenged-murine melanoma B16 cells. BGK-1 which is comparatively more effective than GK-1 extract shows significant suppression of the nuclear factor (NF)-κB activation and inflammatory target genes, in LPS-stimulated RAW 264.7 cells. Conclusion: These results reported effective whitening and anti-inflammatory of BGK-1 as compared to GK-1.

Effect of Paeonia Lactiflora Pallas on Atopic Dermatitis-Related Inflammation in HaCaT Cell (작약이 HaCaT 세포에서 아토피 피부염 관련 염증 억제에 미치는 영향)

  • Lee, Hye-In;Kim, Eom Ji;Son, Dongbin;Joo, Byung Duk;Sohn, Youngjoo;Kim, Eun-Young;Jung, Hyuk-Sang
    • Korean Journal of Acupuncture
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    • v.39 no.2
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    • pp.43-53
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    • 2022
  • Objectives : Paeonia lactiflora Pallas (PLP) have been reported to have pharmacological effects such as anti-inflammatory and analgesic. However, it is not yet known whether PLP extract has anti-inflammatory effect on HaCaT cells, human keratinocyte. Methods : To confirm the anti-inflammatory effect of PLP on keratinocyte, TNF-𝛼/IFN-𝛾-stimulated HaCaT cells were used. HaCaT cells were pre-treated with PLP for 1h before stimulation with TNF-𝛼/IFN-𝛾. Then HaCaT cells were stimulated with TNF-𝛼/IFN-𝛾 for 24 h, the cells and media were harvested to measure the inflammatory cytokines levels. Granulocyte-macrophage colony stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1), interleukin 1 beta (IL-1𝛽), and TNF-𝛼 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression of thymus and activation-regulated chemokines (TARC), IL-6, and IL-8 were measured by reverse transcription-polymerase chain reaction (RT-PCR). We also investigated the inhibitory mechanism of the mitogen-activated protein kinase (MAPKs) including ERK, JNK, and p38 and nuclear factor-kappaB (NF-𝜅B) by PLP using western blot. Results : PLP did not show cytotoxicity in HaCaT cells. In TNF-𝛼/IFN-𝛾-stimulated HaCaT cells, PLP significantly inhibited the expression of GM-CSF, MCP-1 IL-1𝛽, TNF-𝛼, TARC and IL-6. PLP inhibited the phosphorylation of ERK and translocation of NF-𝜅B into the nucleus. Conclusions : These results indicate that PLP could ameliorate the TNF-𝛼/IFN-𝛾-stimulated inflammatory response through inhibition of MAPK and NF-kB signal pathway. This suggests that PLP could be used beneficial agent to improve skin inflammation.

Anti-inflammatory and anti-allergic effect of soybean extracts produced by organic cultivation (유기농 대두 추출물의 항염증 및 항알레르기 효과)

  • Chung, Eun-Kyung;Seo, Eun-Hye;Park, Jun-Ho;Kim, Young-Nam;Kim, Kyung-Hee;Lee, Byung-Ryong
    • Proceedings of the Korean Society of Organic Agriculture Conference
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    • 2009.12a
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    • pp.103-113
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    • 2009
  • This present study was carried out to investigate the biological effects of soybean extracts comparing organic and conventional cultivation. Cellular and molecular analysis was performed to determine anti-oxidative, anti-inflammatory, anti-apoptotic, and anti-inflammatory effects of both soybean extracts. First, we obtained various solvent extracts of soybeans such as water, ethanol, and methanol. Molecular and cellular analysis were performed with 0.1 mg/ml concentration of each solvent extracts. The results of anti-oxidative, anti-inflammatory and anti-apoptotic effects of organic cultivated soybean extracts were prominent than conventional cultivated soybean extracts. However, discrepancy between organic and conventional cultivated soybean extracts was not observed in anti-allergic effects determined by releasing histamine from rat mast cell line, RBL-2H3. Conclusively, organic cultivated soybeans have stronger effects than conventional cultivated soybeans in suppression of inflammation. In addition, organic soybeans could be applied as a functional food ingredient for treatment of chronic inflammation, asthma, and atopic dermatitis with enhanced anti-inflammatory activities.

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Mechanisms of Resorcinol Antagonism of Benzo[a]pyrene-Induced Damage to Human Keratinocytes

  • Lee, Seung Eun;Kwon, Kitae;Oh, Sae Woong;Park, Se Jung;Yu, Eunbi;Kim, Hyeyoun;Yang, Seyoung;Park, Jung Yoen;Chung, Woo-Jae;Cho, Jae Youl;Lee, Jongsung
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.227-233
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    • 2021
  • Benzo[a]pyrene (B[a]P) is a polycyclic aromatic hydrocarbon and ubiquitous environmental toxin with known harmful effects to human health. Abnormal phenotypes of keratinocytes are closely associated with their exposure to B[a]P. Resorcinol is a component of argan oil with reported anticancer activities, but its mechanism of action and potential effect on B[a]P damage to the skin is unknown. In this study, we investigated the effects of resorcinol on B[a]P-induced abnormal keratinocyte biology and its mechanisms of action in human epidermal keratinocyte cell line HaCaT. Resorcinol suppressed aryl hydrocarbon receptor (AhR) activity as evidenced by the inhibition of B[a]P-induced xenobiotic response element (XRE)-reporter activation and cytochrome P450 1A1 (CYP1A1) expression. In addition, resorcinol attenuated B[a]P-induced nuclear translocation of AhR, and production of ROS and pro-inflammatory cytokines. We also found that resorcinol increased nuclear factor (erythroid-derived 2)-like 2 (Nrf2) activity. Antioxidant response element (ARE)-reporter activity and expression of ARE-dependent genes NAD(P)H dehydrogenase [quinone] 1 (NQO1), heme oxygenase-1 (HO-1) were increased by resorcinol. Consistently, resorcinol treatment induced nuclear localization of Nrf2 as seen by Western analysis. Knockdown of Nrf2 attenuated the resorcinol effects on ARE signaling, but knockdown of AhR did not affect resorcinol activation of Nrf2. This suggests that activation of antioxidant activity by resorcinol is not mediated by AhR. These results indicate that resorcinol is protective against effects of B[a]P exposure. The mechanism of action of resorcinol is inhibition of AhR and activation of Nrf2-mediated antioxidant signaling. Our findings suggest that resorcinol may have potential as a protective agent against B[a]P-containing pollutants.

The study of anti-inflammatory effect of Hyeonto-dan extract in RAW 264.7 macrophage (현토단(玄兎丹)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구)

  • Kim, Ma-Ryong;Kang, Ok-Hua;Kong, Ryong;Seo, Yun-Soo;Zhou, Tian;Kim, Sang-A;Kim, Eun-Su;Sin, Min-A;Lee, Young-Seob;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.77-85
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    • 2017
  • Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.

Effects of Chamaeneron angustifolia Extract on Lipid Metabolism and Differentiation of 3T3-L1 Preadipocyte (Chamaeneron angustifolia 추출물이 3T3-L1 Preadipocyte의 지방대사 및 분화에 미치는 영향)

  • Seul Bi Lee;Moon-Yeol Choi;Mi Hyung Kim;So-Young Kim;Mi Ryeo Kim
    • The Korea Journal of Herbology
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    • v.39 no.4
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    • pp.29-35
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    • 2024
  • Objectives : Obesity is an imbalance between energy intake and consumption due to overeating and lack of exercise, and if it persists, it increases non-infectious diseases such as high blood pressure, diabetes, and hyperlipidemia. In this study, we tried to investigate the possibility of using Chamaeneron angustifolia (CA) as a material for anti-obesity by confirming the effect of inhibiting lipid differentiation. Methods : We measured the effects of CA extract on oil-red-o staining, cell cytotoxicity evaluation activity using 3T3-L1 cells. Additionally, we assessed fat decomposition and metabolism-related protein expression through Western blot analysis. Results : In this study, the anti-obesity effects of CA extract were experimentally assessed. Results showed significant inhibition of adipocyte differentiation and accumulation at concentrations of 0.05, 0.1, and 0.2 mg/ml of oil-red-o staining, with reductions of 80% or more. CA notably increased the phosphorylation of AMPK protein expression compared to the control group across all concentrations. Additionally, phosphorylation of ACC significantly increased at a concentration of 0.2 mg/ml compared to the control. PPAR-γ, which regulates adipogenesis, exhibited a significant decrease compared to the control, while protein expression of CPT-1, involved in fatty acid oxidation, showed a concentration-dependent increase across all groups. Therefore, CA extract demonstrates potential as a functional material for anti-obesity by increasing the expression of proteins related to fat decomposition and synthesis while decreasing others. Conclusions : These results suggest that CA may also be useful as an anti-obesity functional substance.