Ethanol Extract of Sarcodon asparatus Mitigates Inflammatory Responses in Lipopolysaccharide-Challenged Mice and Murine Macrophages

정민유,정성근,이혜진,손동화,김현구 한국식품영양과학회 2015 Journal of medicinal food Vol.18 No.11

A number of compounds isolated from mushrooms have exhibited disease-modifying effects. We sought to investigate the mechanisms responsible for the anti-inflammatory effects of an extract from the mushroom species Sarcodon asparatus (SAE). Male BALB/c mice (N = 42; 6 weeks old) were randomly assigned to four treatment groups. Intraperitoneal administration of SAE significantly attenuated lipopolysaccharide (LPS)-mediated increases in alanine aminotransferase (ALT) activity. LPS also increased serum levels of proinflammatory cytokines, including IL-1β, IL-6, and TNF-α, which were dose-dependently and significantly attenuated by SAE. Correlative relationships between serum ALT activity and proinflammatory cytokines suggested that SAE-mediated suppression of liver injury was partly attributable to the attenuation of serum inflammatory responses. SAE significantly decreased hepatic NO· production and subsequent 3-nitrotyrosine formation, and the hepatic NO· production significantly correlated with serum ALT and cytokine levels, suggesting that SAE mitigates liver injury in association with inflammatory processes, likely by suppressing NO· production. Anti-inflammatory activity and further mechanisms of SAE were evaluated using RAW264.7 with LPS challenge. Noncytotoxic levels of SAE significantly attenuated NO· production in RAW264.7 cells and also markedly suppressed the expression of iNOS and other proinflammatory mediators, including COX-2 and IL-6, which were upregulated in the presence of LPS. SAE inhibited the phosphorylation of p65, an observation that occurred independently of IKKαβ-mediated IκBα phosphorylation. Collectively, our results demonstrate that SAE suppressed NO·-mediated inflammation by inhibiting p65 transcriptional activation without affecting IKKαβ-mediated IκBα phosphorylation. Further studies are warranted to examine the major compounds responsible for these effects and the mechanisms responsible for the p65 phosphorylation observed.

Green Tea Lowers Hepatic COX-2 and Prostaglandin E2 in Rats with Dietary Fat-Induced Nonalcoholic Steatohepatitis

정민유,노상규,Eunice Mah,Christopher Masterjohn,박해진,Richard M. Clark,박영기,이지영,Richard S. Bruno 한국식품영양과학회 2015 Journal of medicinal food Vol.18 No.6

Green tea extract (GTE) protects against nonalcoholic steatohepatitis (NASH) by decreasing hepatic steatosis and nuclear factor kappa B (NFκB) activation. We hypothesized that hypolipidemic and anti-inflammatory activities of GTE would protect against NASH by reducing cyclooxygenase-2 (COX-2), an NFκB-dependent enzyme, and prostaglandin E2 (PGE2) in a dietary fat-induced obese model. Male Wistar rats were fed a low-fat diet containing no GTE or a high-fat (HF) diet containing GTE at 0%, 1%, or 2% for 8 weeks. Insulin resistance and total hepatic fatty acids increased following HF feeding (P < .05) and these were normalized by GTE at 1–2%. GTE (1–2%) normalized hepatic malondialdehyde without affecting cytochrome P450 2E1 mRNA expression, which was otherwise increased by HF feeding. HF-mediated increases in hepatic COX-2 protein and activity as well as PGE2 concentrations were normalized by GTE (1–2%). COX-2 activity and PGE2 were correlated to each other, and to serum alanine aminotransferase (ALT) and hepatic NFκB-binding activity (P < .05; r = 0.28–0.49). GTE attenuated HF-mediated increases in total hepatic n-6 and n-3, without affecting the n-6/n-3 ratio. GTE did not affect HF-mediated increases in n-6 in nonesterified fatty acid (NEFA) and phospholipid pools, whereas n-3 and n-6/n-3 in both pools were unaffected by GTE and HF feeding. GTE decreased total hepatic arachidonic acid without affecting HF-mediated increases in arachidonic acid in NEFA or phospholipid pools. Thus, GTE attenuates lipid peroxidation and PGE2 accumulation by decreasing COX-2 activity independent of arachidonic acid availability and supports an additional mechanism by which GTE protects against liver injury during NASH in an HF-feeding model.

Black Mulberry Extract Elicits Hepatoprotective Effects in Nonalcoholic Fatty Liver Disease Models by Inhibition of Histone Acetylation

정민유,김효진,최효경,박재호,황진택 한국식품영양과학회 2021 Journal of medicinal food Vol.24 No.9

Epigenetic regulation by histone acetyltransferase (HAT) is associated with various biological processes and the progression of diseases, including nonalcoholic fatty liver disease (NAFLD). The objective of this study was to investigate whether the hypolipidemic properties of black mulberry (Morus atropurpurea Roxb.) fruit extract (BME) contribute toward protection against NAFLD by HAT inhibition. HepG2 cells were treated with oleic and palmitic acids to induce lipid accumulation, which was significantly attenuated by the treatment with BME at 50 and 100 μg/mL. BME also markedly reduced the expression of proteins associated with lipogenesis, which was attributed to the BME-mediated downregulation of lipogenic genes in HepG2 cells. BME significantly inhibited in vitro total HAT and p300 activities. In addition, BME suppressed total acetylated lysine as well as specific histone acetylation of proteins H3K14 and H3K27 in HepG2 cells. Mice were then fed with either a chow diet or western diet (WD), with or without BME (1%, w/w) supplementation, for 12 weeks to confirm hypolipidemic activity of BME. BME attenuated serum nonesterified fatty acids and low-density lipoprotein (LDL) cholesterol levels, which was likely associated with the downregulation of hepatic lipogenic gene expression in WD-fed obese mice. Taken together, the hypolipidemic activity of BME was observed in HepG2 cells treated with fatty acids as well as in livers of obese mice, and the hepatoprotection of BME is likely associated with the inhibition of acetylation. Further investigation is warranted to determine whether BME can be developed into an efficacious dietary intervention to attenuate the progression of NAFLD by epigenetic regulation in clinical settings.

모시풀 추출물이 지방세포분화와 혈관신생에 미치는 영향

정민유(Min-Yu Chung),김성희(Sung Hee Kim),최효경(Hyo-Kyoung Choi),박재호(JaeHo Park),황진택(Jin-Taek Hwang) 한국생물공학회 2016 KSBB Journal Vol.31 No.3

Boehmeria nivea (L.) Gaud., a flowering plant, has been widely cultivated in Asian countries including Korea. It has been reported that B. nivea exhibits health beneficial effects for the prevention of inflammation, oxidative stress, and virusrelated diseases. In this study, we evaluated the inhibitory effect of B. nivea on adipocyte differentiation and angiogenesis. DPPH radical scavenging activities of 70% ethanol extract of B. nivea (EBN) and water extract of B. nivea (WBN) were 90.8±1.1% and 20±6.9%, respectively. EBN was also effective in the reduction of adipocyte differentiation in 3T3-L1 cells. We next examined the transcriptional activity of peroxisome proliferator-activated receptor gamma (PPAR-γ), a pivotal target for anti-obesity. We found that treatment with rosiglitazone induced the transactivation of PPAR-γ. Under the same condition, 800 μg/mL EBN reduced the transactivation of PPAR-γ in rosiglitazone-induced cells. These results demonstrate that EBN-inhibited adipocyte differentiation was accompanied by PPAR-γ inhibition. The study also tested whether EBN exhibits an anti-angiogenic effect by inhibiting tube formation in HUVECs. We found that EBN effectively inhibits tube formation, suggesting that EBN exhibited an anti-angiogenic effect. Taken together, B. nivea can be used as a functional food for the prevention of obesity and angiogenesisrelated diseases including cancer.

3T3-L1 세포에서 Sodium Selenite의 후성유전 조절 효소 활성 억제를 통한 항비만 효능

이재인,정민유,박선경,홍설민,김영수,박재호 한국식품영양과학회 2023 한국식품영양과학회지 Vol.52 No.3

Selenium is an essential trace mineral for various functions in the body and is known as an antioxidant. To evaluate the anti-obesity effect of sodium selenite (SS), we examined the expressions of epigenetic regulatory enzymes and the anti-adipogenic and lipogenic effects of SS in MDI-induced 3T3-L1 preadipocytes. SS significantly inhibited protein arginine methyltransferase 5 (PRMT5) and histone acetyltransferase (HAT) activity compared with a control, epigallocatechine gallate. SS also attenuated lipid accumulation and triglyceride formation in 3T3-L1 adipocytes. In addition, SS decreased the protein and mRNA levels of adipogenic transcription factors such as peroxisome proliferator-activated receptor gamma, CCAAT/enhaner binding protein alpha. SS effectively suppressed the mRNA expressions of the fatty acid synthase and ATP citrate lyase known as the lipogenic markers and the P300/CBP-associated factor known as an epigenetic regulatory marker in 3T3-L1 adipocytes. Consequently, the anti-obesity effect of SS is likely attributed to the inhibition of PRMT5 and HAT activity. 본 연구에서는 SS의 후성유전 조절 효소인 PRMT5와 HAT 활성의 억제를 통한 항비만 효과를 확인하기 위해 3T3-L1 지방세포에 MDI를 처리하여 지질축적 및 TG 억제능을 평가하였다. SS는 세포독성에 영향을 미치지 않는 농도인 10 μM까지 처리했으며 지질과 TG 농도를 유의하게 억제하였다. 따라서 SS는 항비만 효능이 있음을 확인하였다. In vitro PRMT와 HAT 활성 억제능을 측정하기 위해 EGCG를 대조군으로 사용하여 평가하였다. PRMT와 HAT 활성 효소 억제능은 SS 처리가 EGCG에 대비하여 각각 62%, 97% 억제하였다. 또한 3T3-L1 지방세포에서 specific HAT인 PCAF, CBP, p300의 유전자 발현량을 qRT-PCR로 확인한 결과, PCAF의 발현량이 SS 10 μM 처리에서 현저히 감소하였다. 3T3-L1 지방세포에서 adipogenesis에 관여하는 인자인 PPARγ와 C/EBPα, lipogenesis에 관여하는 인자들(ACLY, ACC, FAS)의 단백질과 유전자 발현을 확인하였다. 그 결과 SS 10 μM 처리에서 adipogenesis 및 lipogenesis 관련 인자들의 단백질과 유전자 발현이 유의하게 억제됨을 확인하였다. 따라서 SS는 PRMT5와 HAT 활성 억제를 통해 항비만 효능을 나타낸다고 할 수 있다.

Conjugated Linoleic Triacylglycerols Exhibit Superior Lymphatic Absorption Than Free Conjugate Linoleic Acids and Have Antiobesity Properties

우현주,정민유,김주연,공대철,민진영,최희돈,최인욱,김인환,노상규,김병희 한국식품영양과학회 2016 Journal of medicinal food Vol.19 No.5

This study aimed to compare lymphatic absorption of conjugated linoleic acids (CLAs) in the triacylglycerol (TAG) or free fatty acid (FFA) form and to examine the antiobesity effects of different doses of CLAs in the TAG form in animals. Conjugated linoleic TAGs (containing 70.3 wt% CLAs; CLA-TAG) were prepared through lipase-catalyzed esterification of glycerol with commercial CLA mixtures (CLA-FFA). Lymphatic absorption of CLA-TAG and CLA-FFA was compared in a rat model of lymphatic cannulation. Greater amounts of cis-9,trans-11 and trans-10,cis-12 CLAs were detected in the collected lymph from a lipid emulsion containing CLA-TAG. This result suggests that CLA-TAG has greater capacity for lymphatic absorption than does CLA-FFA. The antiobesity efficacy of CLA-TAG at different doses was examined in mice with diet-induced obesity. A high-fat diet (HFD) for 12 weeks caused a significant increase in body weight and epididymal and retroperitoneal fat weights, which were significantly decreased by 2% dietary supplementation (w/w) with CLA-TAG. CLA-TAG at 2% significantly attenuated the HFD-induced upregulation of serum TAG, but led to hepatomegaly and exacerbated HFD-induced hypercholesterolemia. CLA-TAG at 1% significantly attenuated upregulation of retroperitoneal fat weight and significantly increased liver weight, which was decreased by the HFD. Nonetheless, the liver weight in group ‘‘HFD +1% CLA-TAG’’ was not significantly different from that of normal diet controls. CLA-TAG at 1% significantly reduced serum TAG levels and did not exacerbate HFD-induced hypercholesterolemia. Thus, 1% dietary supplementation with CLA-TAG reduces retroperitoneal fat weight without apparent hepatomegaly, a known side-effect of CLAs in mouse models of obesity.

Anacardic Acid Suppresses Adipogenesis Through Inhibition of the Hsp90/Akt Signaling Pathway in 3T3-L1 Preadipocytes

이장호,정민유,정상원,최효경 한국식품영양과학회 2021 Journal of medicinal food Vol.24 No.5

Anacardic acid (AA), a major component of cashew nut shell liquid, has extensive bioactivities. However, little is known about its antiadipogenic properties or the mechanism that underpins them. The aim of this study was to investigate the effect of AA on 3T3-L1 preadipocyte differentiation and its mechanisms of action. AA inhibits lipid accumulation during adipogenesis in 3T3-L1 preadipocyte (IC50 = 25.45 μM). AA abrogates mRNA expressions of the genes implicated in lipogenesis and their transcription factors, especially Pparg and Cebpa. Furthermore, antibody microarray and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis results showed that the proteins implicated in the Akt signaling pathway were most likely altered by AA. Notably, upon AA treatment, heat shock protein 90 (Hsp90), a positive regulator of Akt, was decreased, resulting in Akt degradation. These findings indicate that AA, a natural product that acts as a Hsp90/Akt signaling inhibitor, may be a possible antiadipogenic agent.

흑미 및 검은콩 섭취가 한국 성인의 대사증후군 요인에 미치는 영향

정상원,정민유,최효경,박재호,황진택 한국식품영양과학회 2017 한국식품영양과학회 학술대회발표집 Vol.2016 No.10

Ethanol Extract of Ligularia fischeri Inhibits the Lipopolysaccharide-Induced Inflammatory Response by Exerting Anti-Histone Acetyltransferase Activity to Negatively Regulate p65

김효진,최효경,정민유,박재호,정상원,이승현,황진택 한국식품영양과학회 2019 Journal of medicinal food Vol.22 No.11

Histone acetyltransferase (HAT) activity is well established to regulate inflammatory responses. In contrast, the mechanisms by which natural nutritional extracts influence epigenetic mechanisms to regulate inflammation have not yet been thoroughly investigated. Thus, in the present study, we observed that the anti-HAT activity exerted by an ethanol extract of Ligularia fischeri (ELF) inhibited inflammation. Specifically, we used a cell-free system to show that ELF attenuates HAT activity. We also demonstrated that ELF decreases lipopolysaccharide (LPS)-induced HAT mRNA and protein expression levels in Raw 264.7 cells, and thereby attenuates inflammation-induced patterns of hyperacetylation at nonhistone and histone-H4 proteins. Interestingly, we found that ELF blocked p65 translocation in LPS-stimulated Raw 264.7 cells by attenuating acetylation at lysine residue 310 of p65. Finally, we investigated whether ELF reduces the inflammatory cytokines, IL-6, IL-1β, and TNFα, using its HAT inhibitor activity. Taken together, these results suggest that ELF negatively regulates inflammatory responses by inhibiting HATs and HAT activity.

RAW264.7 세포에서 새싹보리 및 Luteolin의 히스톤 아세틸기전달효소 활성 억제를 통한 항염증 효능

이재인(Jae-In Lee),정민유(Min-Yu Chung) 한국식품영양과학회 2021 한국식품영양과학회지 Vol.50 No.12

새싹보리의 항염증 효능을 확인하기 위하여 RAW264.7 마우스 대식세포에서 LPS에 의한 NO 생성 억제 정도를 평가하였다. BSP, BSE, BSW 중 BSE와 BSW가 cell viability에 영향을 미치지 않는 농도에서 NO 생성을 억제하였다. 새싹보리 열수 및 에탄올 추출물 모두 항염증 효능이 있음을 확인하였다. 새싹보리의 주요 활성 성분 중 luteolin의 NO 생성 및 in vitro HAT 활성 억제능이 다른 성분들보다 뛰어났다. LPS로 염증반응이 유도된 RAW264.7 세포에서 luteolin은 염증매개인자(IL-6, TNF-α, iNOS, COX-2)의 발현을 감소시켰다. Luteolin의 total 및 specific HAT 활성 조절 정도를 평가하였는데, total HAT 활성에 있어서 luteolin의 IC50은 48.57 μM이었다. Luteolin은 또한 저농도 처리(0.1 μM)에서부터 p300, CBP, PCAF의 활성을 현저하게 감소시켰다. Luteolin의 HAT 활성 억제능은 LPS로 염증반응이 유도된 RAW264.7 세포의 핵 내에서도 동일하게 관찰되었다. Luteolin의 항염증 효능은 HAT 활성 억제와 연관이 있으며, 새싹보리에서 관찰된 항염증 효능은 luteolin의 HAT 억제를 통한 항염증 효능에 기인한다고 할 수 있겠다. This study aimed to assess the anti-inflammatory activity and the mechanism of anti-inflammatory action of barley sprout (Hordeum vulgare L.). Barley sprout powder (BSP), barley sprout ethanol extract (BSE), and barley sprout water extract (BSW) were prepared, and the inhibitory effects of the samples on nitric oxide (NO) production were evaluated. The results showed that the inhibitory activity of BSE and BSW on NO production was greater compared to BSP. Among the active compounds found in barley sprouts, luteolin exhibited greater inhibition of NO production and histone acetyltransferase (HAT) activity compared to the other compounds. Luteolin significantly (P<0.05) attenuated lipopolysaccharide (LPS)-mediated inflammatory mediators, including interleukin-6, tumor necrosis factor-α, inducible nitric oxide synthase, and cyclooxygenase-2 in RAW264.7 cells. Luteolin also significantly inhibited in vitro specific HATs (p300, CBP, PCAF) as well as total HAT, which was also observed in the nuclear fraction of LPS-treated RAW264.7 cells. The anti-inflammatory activity of luteolin was attributed to total and specific HAT inhibition. Collectively, the anti-inflammatory activity of barley sprouts could be attributed to luteolin via HAT inhibition.