Effects of Raphanus sativus var. niger (Black Radish) Fermented with Lactobacillus plantarum on Alcohol Metabolism in Rats

강경구,Seo Min-Soo,Sung Soo-Eun,Choi Joo-Hee,Lee Si-Joon,Kim KilSoo,Jang Wookju,Lee Hak Sung 건강기능식품미래포럼 2021 건강기능식품미래포럼 학술지 Vol.1 No.2

Excessive drinking causes various side effects including hepatic and neurological diseases that can lead to serious social problems, and thus, efforts to search for a way to help to promote alcohol decomposition are increasingly needed. Raphanus sativus var. niger commonly known as black radish has been reported to have biologically active glycosides such as glucosinolates and also known to improve liver functions, suggesting that this vegetable may have an action to promote alcohol metabolism. In the present study, this possibility was tested with an extract prepared from the black radish fermented with Lactobacillus plantarum in rats. The fermented black radish extract (75 or 300 mg/kg) or saline (2 mL/kg) was administered to rats orally. After 30 minutes, ethanol (4 g/kg) was administered orally using 25% ethanol in water and blood was sampled at 0, 1, 2 and 4 hours after ethanol administration and then euthanasia was performed to get the liver. The sera obtained from the blood samples were used for the concentration analysis of ethanol and acetaldehyde and the examination of blood chemistries. The liver tissues were used for the activity assays for alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). It was observed that in the rats administered the fermented black radish extract, the concentrations of ethanol and acetaldehyde decreased more and simultaneously the activities of ADH and ALDH increased more than those in the saline-treated rats. Any significant changes in the blood chemistries and histology of liver tissues were not observed. These results suggest that the fermented black radish extract has a potential that may ameliorate hangover symptoms caused by excessive alcohol ingestion.

유산균 발효 흑무의 항산화 및 지방구 형성 억제 효과

김성은(Seong-Eun Kim),백신화(Sinhwa Baek),이학성(Hak Sung Lee),김현규(Hyun-Kyu Kim) 한국식품영양과학회 2018 한국식품영양과학회지 Vol.47 No.7

본 연구는 Lactobacillus plantarum을 이용하여 발효시킨 흑무 유산균 발효물의 간 손상 보호 효과를 알아보기 위해 기초적인 항산화 효능 및 간세포 지방구 형성 억제 활성 실험을 하였다. 흑무 유산균 발효물은 단순 추출물 및 발효 전 단계의 영양원과 비교하여 유의적으로 증진된 항산화 효능을 보였으며, 총 폴리페놀 함량 및 총 플라보노이드 함량 역시 발효를 통해 증가한 것을 확인하였다. 또한, 간 손상 유발 시 간세포에 형성되는 지방구의 형성 억제를 통한 간세포 보호 효능도 단순 추출물 및 발효 전 단계 영양원과 대비하여 유의적으로 증가한 것을 확인하였다. 이상의 결과를 통해 흑무 유산균 발효물은 종전의 흑무 추출물과 대비하여 손상에 의한 간세포의 지방구 형성 억제 효능이 증진되었으며 이를 토대로 유산균 발효 흑무는 간 건강 개선을 위한 건강기능식품 소재로써 활용될 수 있는 가능성을 확인하였다. Black radish (Raphanus sativus var. niger) has been reported to have anti-oxidant, anti-inflammatory and detoxification effects. In this study, we investigated the antioxidant and lipid accumulation inhibitory effects of fermented black radish (FBR) with Lactobacillus plantarum compared with non-fermented black radish (BR) and black radish extracts obtained using water, and 30%, 50%, 70% ethyl alcohol. The lipid accumulation inhibitory effect was determined by Oil-Red O staining activity after oleic acid treatment of HepG2 cells. Treatment with FBR at doses of 100 and 500 μg/mL inhibited lipid accumulation by 103% and 108% and BR at the same dose inhibited lipid accumulation by 72% and 61%, respectively. Extracts inhibited lipid accumulation ranging by 17% to 93% when applied in the same doses. FBR led to significant inhibition of lipid accumulation compared with BR and extracts. In addition, FBR demonstrated antioxidant effect such as scavenging activity of (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl radical and super oxide dismutase (SOD)-like activity. Also the FBR was measured total polyphenol contents (TPC) and total flavonoid contents (TFC). Moreover, FBR showed significant greater antioxidant activity than BR and extracts. The TPC of FBR increased more than BR and the TFC of FBR was higher than that of other extracts. These results suggest that FBR might be a potential health functional food ingredient for enhancing liver function.

Antimelanogenic Effects of Raphanus sativus L. var. niger Roots on α-MSH Stimulated B16F10 Melanoma Cells

고하나,김정은,조연정,홍승현,양다운,김기옥,이남호 대한화학회 2018 Bulletin of the Korean Chemical Society Vol.39 No.11

This study was conducted to identify the antimelanogenic components from Raphanus sativus L. var. niger (black radish). Phytochemical study with the ethanol extract of black radish roots resulted in the isolation of six compounds: 3-(E)-(methylthio)methylene-2-pyrrolidinethione (1), ascorbigen (2), ?-sitosterol (3), palmitic acid (4), α-linolenic acid (5), and 1-linoleonyl glycerol (6). Among the isolates, the thione 1 efficiently inhibited melanin synthesis in a concentration-dependent manner in α-MSH stimulated B16F10 melanoma cells. Western blotting analysis for the key melanogenic enzymes showed that compound 1 decreased tyrosinase and tyrosinase-related protein-2 expressions. These results demonstrated that the black radish extract including pyrrolidinethione 1 could be useful as antimelanogenic agents in cosmetic or pharmaceutical applications.

Inhibitory Effect of Black Radish (Raphanus sativus L. var. niger) Extracts on Lipopolysaccharide-Induced Inflammatory Response in the Mouse Monocyte/Macrophage-Like Cell Line RAW 264.7

Hyungsik Jeon,Dawun Yang,Nam Ho Lee,Meejung Ahn,Giok Kim 한국식품영양과학회 2020 Preventive Nutrition and Food Science Vol.25 No.4

Black radish (Raphanus sativus L. var. niger), which is cultivated worldwide, is used in traditional medicine as it aids liver function, gastric secretion, gallbladder function, and gallstone mitigation. In this study, we examined the anti-inflammatory effects of black radish extract (BRE) on the lipopolysaccharide (LPS)- and interleukin (IL)-6-mediated inflammatory responses in the RAW 264.7 cell lines. Our findings show that BRE significantly ameliorated LPS-induced nitric oxide (NO) release and production of pro-inflammatory cytokines, such as IL-1β, IL-6, tumor necrosis factor (TNF)-α,and prostaglandin E2. The levels of cyclooxygenase (COX)-2 and inducible NO synthase (iNOS) in LPS-stimulated RAW 264.7 cells were found to be suppressed by BRE. Further, BRE significantly suppressed the LPS-induced expression of mRNAs encoding COX-2, iNOS, IL-1β, IL-6, and TNF-α in a concentration-dependent manner. BRE treatment significantly inhibited Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) phosphorylation in IL-6- and LPS-treated RAW 264.7 cells. In addition, BRE decreased the levels of phosphorylated extracellular signal-regulated protein kinases and c-Jun N-terminal kinase under the same conditions. Moreover, BRE induced high nuclear factor erythroid 2-related factor 2 (NRF2) levels and its target gene heme oxygenase 1 (HO-1) in the absence of LPS. These data demonstrate that BRE may be beneficial for treating inflammation through selective immunomodulatory effects, which may be mediated by inhibition of the STAT3/JAK2 and activation of the NRF2/HO-1 signal transduction pathways.

Hepatoprotective effect of fermented black radish (Raphanus sativus L. var niger) in CCl₄ induced liver injury in rats

Jeong tae Kim,Mee jung Ahn,Seong Eun Kim,Hak Sung Lee,Hyun Kyu Kim,Gi Ok Kim,Tae kyun Shin 한국예방수의학회 2017 예방수의학회지 Vol.41 No.4

Oxidative stress is one of common cause of fatty changes in the liver. Antioxidant capacity was confirmed in various vegetables including black radish (Raphanus sativus L. var niger). Fermentation of vegetables using Lactobacillus plantarum has been known to generate bioactive components. This study was conducted to determine if fermented black radish (FBR) ameliorates oxidative liver injury induced by CCl4 in rats. To accomplish this, FBR (250 and 500 mg/ kg) was orally administered to rats for 7 consecutive days, single CCl₄ (1.5 mL/kg) treatment or no treatment orally. Serum chemistry at 24 hours after CCl₄ injury showed that FBR (500 mg/kg) significantly reduced the level of both alanine aminotransferase and aspartate aminotransferase in CCl₄ exposed rats. Moreover, FBR treatment significantly increased radical-scavenging effects in livers with the reduction of lipid peroxidation in CCl₄ exposed rats. Histopathologic findings including Kupffer cell activation in the liver of each group matched those of serum chemistry. Collectively, black radish, through fermentation, exerts hepatoprotective capacity in CCl₄ induced liver injury in rats through anti-oxidation.

다양한 추출 및 건조방법으로 제조된 흑무 추출물의 특성

김하영,신태균,안미정,김기옥,천지연 공주대학교 자원과학연구소 2021 자원과학연구 Vol.3 No.1

This study investigated the effect of the extraction method and drying method on black radish quality properties. The yellowness of black radish extracts was significantly different according to the extraction and drying method (p<0.05). The yield was higher at L.plantarum fermentation and freeze-dried extracts, and there was no difference depending on inlet temperature and extraction method. The extracts by ethanol extraction and spray-drying were smallest particle size (107-118 nm), highest zeta-potential (–33-–39 mV), and low polydispersity index (0.238-0.250). In total phenol contents and Ferric reducing antioxidant power evaluation, spray-dried ethanol extracts were higher total phenol contents and L. plantarum fermentation and freeze-dried extracts showed the highest ferric reducing antioxidant power. It was suggested that spray-dried black radish ethanol extracts were economically effective in applying the food process and were expected to be bioavailability in the digestion system because of micronization by spray-drying.

3T3-L1 세포에서 검정무 추출물의 Adipogenesis 억제 효과

양다운,홍승현,이학성,이남호,신태균,김기옥 한국식품영양과학회 2019 한국식품영양과학회지 Vol.48 No.8

Black radish (Raphanus sativus L. var. niger) is a root vegetable that is distributed all over the world and it is used for folk medicine to treat jaundice, liver disease, indigestion and gastric pains. In the present study, we investigated the anti-obesity activity of black radish extract (BRE) by conducting various experiments. The inhibitory, dose dependent effect of BRE on adipogenesis was shown in 3T3-L1 preadipocytes with Oil Red O staining, and it effectively decreased the intracellular triglyceride concentration of adipocytes. The effect of BRE on the expression of various adipogenesis-related genes was analyzed via western blotting and quantitative real time PCR. We demonstrated that BRE significantly down regulated the expression of C/EBPα, C/EBPδ, PPARγ, aP2, SREBP-1c, ADD1, ACC, FAS, SCD, ATGL, HSL, and adipsin in a dose dependent manner. The specific mechanism mediating the anti-adipogenesis effects of BRE was the inhibitory effect of BRE on AMP-activated protein kinase activation and the Nrf2/HO-1 pathway. In addition, we found that BRE significantly increased C/EBPβ protein and mRNA, and this was all influenced through Nrf2 activation during an early stage (day 2). BRE inhibits lipid accumulation by regulating adipogenesis and lipogenesis related genes and signaling proteins. BRE may a good candidate for a nutraceutical and pharmaceutical formulation for treating or controlling obesity. 본 연구에서는 검정무 추출물의 항비만 효과 및 그에 따른 생화학적과 분자생물학적 기전 연구를 MDI를 처리한 3T3- L1 세포에서 adipogenesis 과정에 관여하는 전사인자(C/ EBPs, SREBP-1c, ADD1, PPARγ), 지방대사 관련 유전자(aP2, ACC, FAS, SCD, ATGL, HSL, adipsin)와 AMPK/ Nrf2 pathway를 분화 후 day 2~day 8에서 western blot과 qPCR 기법으로 분석하였다. 3T3-L1 세포에서 검정무 추출물은 lipid droplet 형성과 triglyceride 축적을 현저하게 감소시키는 결과를 관찰하였다. 또한 검정무 추출물을 분화 후 day 2와 day 6에 처리한 결과 adipogenesis의 전사조절인자와 단백질인 C/EBPα, PPARγ, aP2의 단백질 발현 수준을 감소시켰다. 그리고 분화시기별로 phospho-AMPKα와 PPARγ 발현을 분석한 결과, 검정무 추출물은 AMPK 활성화를 통해서 지방합성 핵심인자인 PPARγ를 억제하는 것으로 확인되었다. 그리고 qPCR을 통해 day 2와 day 6에서 전사인자 및 지방대사 관련 유전자 12개(C/EBPα, C/ EBPδ, SREBP-1c, ADD1, PPARγ, aP2, ACC, FAS, SCD, ATGL, HSL, adipsin)를 분석한 결과, 모두 농도 의존적으로 mRNA 발현이 감소하였다. 분화초기 특이적 유전자들을 통해 adipogenesis 억제를 통해 항비만 활성을 나타내었다. 마지막으로 검정무 추출물은 Nrf2와 HO-1 단백질을 농도 의존적으로 증가시켰고 HO-1과 NQO1 mRNA 발현도 증가시킴을 확인하였다. 따라서 검정무 추출물은 직접적으로 C/ EBP 전사인자, PPARγ 발현을 억제할 뿐만 아니라 AMPK를 활성화하고 ACC 단백질을 불활성화함으로써 지방세포분화를 억제하는 것으로 사료된다. 그리고 검정무 추출물은 Nrf2/HO-1 pathway를 활성화하여 adipogenesis 동안 산화스트레스를 해소하여 지방세포분화를 억제하는 것으로 사료된다. 따라서 검정무는 체지방감소 건강기능식품 및 미용기능식품 기능성 소재로서의 활용 가능성이 높다고 생각되며, 향후 전임상 및 임상 연구를 통해 식의약품 소재 개발 연구가 추가로 요구된다.

Hepatoprotective effect of both aqueous extracted and fermented black radish (Raphanus sativus L. var niger) in CCl4 induced liver injury in rats by attenuating oxidative stress

Meejung Ahn,Jeongtae Kim,Seong-Eun Kim,Hak Sung Lee,Hyun Kyu Kim,Seunghyun Hong,Gi Ok Kim,Taekyun Shin 한국실험동물학회 2018 한국실험동물학회 학술발표대회 논문집 Vol.2018 No.1

Black Radish (Raphanus sativus L. var. niger) Extract Mediates Its Hepatoprotective Effect on Carbon Tetrachloride-Induced Hepatic Injury by Attenuating Oxidative Stress

안미정,김정태,홍승현,김정은,고하나,이남호,김기옥,신태균 한국식품영양과학회 2018 Journal of medicinal food Vol.21 No.9

Nonalcoholic fatty liver disease is a serious liver disorder associated with oxidative stress. Black radish (Raphanus sativus L. var. niger) extract (BRE) can lower the risk of this disease. The hepatoprotective effect of BRE containing 3-(E)-(methylthio)methylene-2-pyrrolidinethione was evaluated in human hepatocyte carcinoma (HepG2) cells and in rat livers with carbon tetrachloride (CCl4)-induced hepatic injury. BRE was administered at 125, 250, 500, and 1000 μg/mL to the oleic acid-induced HepG2 cells. Male Sprague-Dawley rats were randomly divided into seven groups: the control group, BRE group, CCl4 group, and BRE + CCl4 group. BRE was administered orally at 125, 250, 500, and 1000 mg/kg/day once daily for 7 consecutive days, followed by a single oral treatment of 1.5 mL/kg CCl4. Inhibition of lipid accumulation, serum markers of liver injury, histological evaluations, levels of oxidative stress related enzymatic and nonenzymatic antioxidants in HepG2 cells and liver tissue were investigated. The protein expression of main liver P450 isoenzymes such as cytochrome p450(CYP)2E1, the expression of nuclear factor erythroid 2-related factor-2(Nrf-2) and heme oxygenase-1(HO-1) were also studied. BRE has an inhibitory effect on lipid accumulation and caused acute hepatotoxicity manifested by increased levels of lipid peroxidation, serum alanine aminotransferase, and aspartate aminotransferase with corresponding histopathological changes and high levels of oxidative stress. BRE treatment significantly increased the level of CYP2E1, Nrf-2, and HO-1 in a dose-dependent manner. Besides, 3-(E)-(methylthio)methylene-2-pyrrolidinethione significantly increased radical-scavenging effects and the expression of Nrf-2 in oleic acid-treated HepG2 cells. These results suggest that BRE treatment reduces lipid accumulation in oleic acid-induced steatosis of HepG2 cells, and has a hepatoprotective effect against CCl4-induced liver injury in rats, possibly through Nrf-2/HO-1-mediated antioxidant effects.

Black radish(Raphanus sativus var. niger) inhibits adipogenesis in 3T3L1 cells

Da Wun YANG,Seung Hyun HONG,Seong Eun KIM,Hak Sung LEE,Gi Ok KIM 한국생물공학회 2018 한국생물공학회 학술대회 Vol.2018 No.10