• Title/Summary/Keyword: Cell BE

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LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma (황련 추출물의 LC-MS/MS 분석 및 항염증 효과)

  • Min-Jung, Kim;Ye-Jin, Yang;Kwang-Youn, Kim;Hun Hwan, Kim;Jae Dong, Son;Ju-Hye, Yang;Dong bin, Lee;Woo Hyun, Kim;Hu-Jang, Lee;Seon Been, Bak;Kwang-Il, Park
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.1-10
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    • 2023
  • Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.

Ginsenoside compound K protects against cerebral ischemia/ reperfusion injury via Mul1/Mfn2-mediated mitochondrial dynamics and bioenergy

  • Qingxia Huang;Jing Li;Jinjin Chen;Zepeng Zhang;Peng Xu;Hongyu Qi;Zhaoqiang Chen;Jiaqi Liu;Jing Lu;Mengqi Shi;Yibin Zhang;Ying Ma;Daqing Zhao;Xiangyan Li
    • Journal of Ginseng Research
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    • v.47 no.3
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    • pp.408-419
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    • 2023
  • Background: Ginsenoside compound K (CK), the main active metabolite in Panax ginseng, has shown good safety and bioavailability in clinical trials and exerts neuroprotective effects in cerebral ischemic stroke. However, its potential role in the prevention of cerebral ischemia/reperfusion (I/R) injury remains unclear. Our study aimed to investigate the molecular mechanism of ginsenoside CK against cerebral I/R injury. Methods: We used a combination of in vitro and in vivo models, including oxygen and glucose deprivation/reperfusion induced PC12 cell model and middle cerebral artery occlusion/reperfusion induced rat model, to mimic I/R injury. Intracellular oxygen consumption and extracellular acidification rate were analyzed by Seahorse multifunctional energy metabolism system; ATP production was detected by luciferase method. The number and size of mitochondria were analyzed by transmission electron microscopy and MitoTracker probe combined with confocal laser microscopy. The potential mechanisms of ginsenoside CK on mitochondrial dynamics and bioenergy were evaluated by RNA interference, pharmacological antagonism combined with co-immunoprecipitation analysis and phenotypic analysis. Results: Ginsenoside CK pretreatment could attenuate mitochondrial translocation of DRP1, mitophagy, mitochondrial apoptosis, and neuronal bioenergy imbalance against cerebral I/R injury in both in vitro and in vivo models. Our data also confirmed that ginsenoside CK administration could reduce the binding affinity of Mul1 and Mfn2 to inhibit the ubiquitination and degradation of Mfn2, thereby elevating the protein level of Mfn2 in cerebral I/R injury. Conclusion: These data provide evidence that ginsenoside CK may be a promising therapeutic agent against cerebral I/R injury via Mul1/Mfn2 mediated mitochondrial dynamics and bioenergy.

Physicochemical Characteristics and Skin Absorption of Transfersomes Containing Centella asiatica Extract According to Edge Activators (Edge Activator 에 따른 병풀추출물 함유 트렌스퍼좀의 물리화학적 특성과 피부흡수)

  • Eun-hee Lee;Kyung-Sup Yoon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.2
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    • pp.147-157
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    • 2023
  • Centella asiatica extract is widely used as a raw material for cosmetics due to its various effects, but it is difficult to expect penetration into the skin due to its high molecular weight and low solubility. In order to solve these problems, lipid-based liposomes of various types were developed to increase skin absorption. Therefore, in this study, we tried to increase the skin absorption rate by preparing transfersomes using surfactants as edge activators in existing liposomes. Liposome and transfersomes containing Span 80 and Tween 20, 60, 80, and 85, respectively, were prepared using a high-pressure homogenizer, and we evaluated the particle size, polydispersity index, zeta potential, and skin absorption rate. As a result, there was almost no change in the physical properties of particle size, polydispersity index and zeta potential from 25 ℃ to 60 d, and the particle size of transfersomes containing Tween 20, 60, and 80 increased after 60 d at 45 ℃. Madecassoside, main substances of the Centella asiatica extract was used as an standard and madecassoside was measured and calculated when measuring the skin absorption rate using Franz diffusion cells. As a result, formulations containing Tween 20 were the most, whereas formulations containing Span 80 were the least. According to the skin absorption coefficient (Kp) value, all formulations showed 'very fast', and the absorption rate was similar or greater than that of liposomes, except for formulations containing Span 80. Through this, it was confirmed that the larger the HLB value of the nonionic surfactant, the smaller the particle size of the transfersome, and the increased skin absorption rate due to the increased flexibility of the vesicle membrane. Through this study, transfersome using surfactant as an edge activator can be expected to solve local skin problems not only as a cosmetic raw material or product, but also by increasing skin absorption.

Anti-inflammatory Effects, Skin Wound Healing, and Stability of Bluish-purple Color Extracted from Platycodon grandiflorus (Jacq.) A.DC. Flower Extract (도라지꽃 추출물의 항염증, 피부재생 효과 및 색소 안정성 연구)

  • Jin-A Ko;Jiwon Han;Bomi Nam;Beom seok Lee;Jiyoung Hwang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.4
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    • pp.313-321
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    • 2023
  • Platycodon grandiflorus (P. grandiflorus) flower is a perennial plant belonging to the family Campanulaceae and has many excellent pharmacological effects, so it has been used as a medicinal ingredient since ancient times. In addition, anthocyanin is a purple or blue natural pigment contained in plant flowers and fruits, and is known as a powerful antioxidant. The purpose of this study was to confirm the dermatological functionality of P. grandiflorus flower extract and the value of the bluish anthocyanin contained in flowers as a cosmetic material as a natural pigment. Firstly, 50% ethanol and 80% ethanol were added to the P. grandiflorus flower and extracted under reflux for 4 h at 25, 60, and 80 ℃, and the pH of each treatment group was similar. Based on the anthocyanin content and chromaticity (E*ab), 50% ethanol 60 ℃ extraction conditions showing the color development most similar to the natural color of the P. grandifloras flower were selected, and a sample was prepared by concentrating and lyophilizing. The analysis results showed that the total phenol, total flavonoid, and total anthocyanin contents were in the ranges of 23 ㎍/mL, 16 ㎍/mL, and 0.17 ㎍/mL, respectively. The P. grandiflorus flower extract suppressed the production of nitric oxide (NO) and interleukin-6 (IL-6) in lipopolysaccharide (LPS) induced RAW264.7 cells. Furthermore, the P. grandiflorus flower extract showed wound healing effects through the promotion of skin cell migration in TNF-α stimulated human keratinocytes. The stability of anthocyanin and extract color was studied during a storage period of 50 days at various temperatures (4 ℃, 25 ℃, and 45 ℃). Color values (L, a, and b) of the P. grandiflorus flower extract changed over 50 days, whereas the bluish-purple color of the extract was stabilized using 5% maltodextrin. These results suggest that P. grandiflorus flower extract may be useful as a natural cosmetic pigment.

Effect of Varying Excessive Air Ratios on Nitrogen Oxides and Fuel Consumption Rate during Warm-up in a 2-L Hydrogen Direct Injection Spark Ignition Engine (2 L급 수소 직접분사 전기점화 엔진의 워밍업 시 공기과잉률에 따른 질소산화물 배출 및 연료 소모율에 대한 실험적 분석)

  • Jun Ha;Yongrae Kim;Cheolwoong Park;Young Choi;Jeongwoo Lee
    • Journal of the Korean Institute of Gas
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    • v.27 no.3
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    • pp.52-58
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    • 2023
  • With the increasing awareness of the importance of carbon neutrality in response to global climate change, the utilization of hydrogen as a carbon-free fuel source is also growing. Hydrogen is commonly used in fuel cells (FC), but it can also be utilized in internal combustion engines (ICE) that are based on combustion. Particularly, ICEs that already have established infrastructure for production and supply can greatly contribute to the expansion of hydrogen energy utilization when it becomes difficult to rely solely on fuel cells or expand their infrastructure. However, a disadvantage of utilizing hydrogen through combustion is the potential generation of nitrogen oxides (NOx), which are harmful emissions formed when nitrogen in the air reacts with oxygen at high temperatures. In particular, for the EURO-7 exhaust regulation, which includes cold start operation, efforts to reduce exhaust emissions during the warm-up process are required. Therefore, in this study, the characteristics of nitrogen oxides and fuel consumption were investigated during the warm-up process of cooling water from room temperature to 88℃ using a 2-liter direct injection spark ignition (SI) engine fueled with hydrogen. One advantage of hydrogen, compared to conventional fuels like gasoline, natural gas, and liquefied petroleum gas (LPG), is its wide flammable range, which allows for sparser control of the excessive air ratio. In this study, the excessive air ratio was varied as 1.6/1.8/2.0 during the warm-up process, and the results were analyzed. The experimental results show that as the excessive air ratio becomes sparser during warm-up, the emission of nitrogen oxides per unit time decreases, and the thermal efficiency relatively increases. However, as the time required to reach the final temperature becomes longer, the cumulative emissions and fuel consumption may worsen.

Kinesin Superfamily Protein 5A (KIF5A) Binds to ArfGAP1, ADP-ribosylation Factor GTPase-activating Protein 1 (Kinesin Superfamily Protein 5A (KIF5A)와 ADP-ribosylation Factor GTPase-activating Protein 1 (ArfGAP1)의 결합)

  • Myoung Hun Kim;Se Young Pyo;Eun Joo Chung;Young Joo Jeong;Sung Woo Park;Mi Kyoung Seo;Won Hee Lee;Sang-Hwa Urm;Mooseong Kim;Dae-Hyun Seog
    • Journal of Life Science
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    • v.34 no.5
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    • pp.333-338
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    • 2024
  • Kinesin-1 is a heterotetrameric protein composed of two heavy chains (KHCs, also known as KIF5s) with a motor domain and two light chains (KLCs) without a motor domain. KIF5 has three subtypes, namely, KIF5A, KIF5B, and KIF5C, which share high amino acid homology except in their carboxy (C)-terminal region. KIF5A is responsible for transporting cargo within the cell. The adaptor proteins that bind to the C-terminal region of KIF5A mediate between kinesin-1 and cargo. However, the proteins regulating the intracellular cargo transport of kinesin-1 have not yet been fully identified. In this study, we identified ADP-ribosylation factor GTPase-activating protein 1 (ArfGAP1), which is involved in the intracellular trafficking of lysosomes, as a binding partner of KIF5A. KIF5A binds to the C-terminal region of ArfGAP1, and ArfGAP1 binds to the C-terminal region of KIF5A but does not interact with KIF5B, KIF5C, kinesin light chain 1 (KLC1), or KIF3A. When co-expressed in mammalian cells, ArfGAP1 co-localized with KIF5A and co-immunoprecipitated with KIF5A, KIF5B, and KLC1, but not with KIF3B. These results suggest that kinesin-1 may be regulated by ArfGAP1 in the intracellular transport of cargo.

L-AHG-mediated Suppression of M1 Polarization and Pro-inflammatory Signaling Pathways in LPS-stimulated RAW264.7 Macrophages (LPS에 의해 자극된 RAW264.7 대식세포에서 L-AHG에 의한 M1 분극화 및 친염증 신호 경로의 억제)

  • Won Young Jang;Shin Young Park;Ki Youn Kim;Do Youn Jun;Young-Seuk Bae;Young Ho Kim
    • Journal of Life Science
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    • v.34 no.7
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    • pp.443-452
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    • 2024
  • This study aimed to examine the influence of 3,6-anhydroxygalactose (L-AHG) on the pro-inflammatory M1 polarization and pro-inflammatory responses observed in the RAW264.7 mouse macrophage cell line following stimulation with lipopolysaccharides (LPS). L-AHG exhibited a significant and dose-dependent inhibition of inducible nitric oxide synthase (iNOS) expression, a hallmark of M1 polarization, and subsequent NO production in LPS-stimulated RAW264.7 cells. Furthermore, the LPS-induced upregulation of cyclooxygenase-2 (COX-2), which drives the production of prostaglandin E2, an inflammatory mediator, was also inhibited by L-AHG. L-AHG did not affect the LPS-triggered Toll-like receptor 4 (TLR4)-mediated pro-inflammatory signaling pathway, which culminated in the activation of transforming growth factor-β-activated kinase 1 (TAK1). However, it was observed to inhibit the generation of reactive oxugen species (ROS) in a dose-dependent manner, as well as the TAK1-driven activation of JNK and p38 MAPK. Given that the active p38 MAPK is known to contribute to the assembly of active nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, which catalyzes the intracellular generation of pro-inflammatory ROS in LPS-stimulated macrophages, the dose-dependent reduction in the LPS-induced ROS generation by L-AHG may be mainly due to the prevention of TAK1-driven activation of p38 MAPK. Together, these results demonstrate that the L-AHG-mediated inhibition of the TAK1-JNK/p38 MAPK activation phase of the pro-inflammatory signaling pathway in LPS-stimulated RAW264.7 cells by L-AHG represents a promising mechanism for suppressing M1 polarization and pro-inflammatory responses in macrophages.

Production of highly enriched GABA through Lactobacillus plantarum fermentation of katsuobushi protein hydrolyzate made from Dendropanax morbiferus extract fermented by Bacillus subtilis (황칠나무 추출물의 고초균 발효물로 제조된 가쓰오부시 단백가수분해물의 Lactobacillus plantarum 발효를 통한 고농도 GABA 생산)

  • Yu-Jeong An;Nak-Ju Sung;Sam-Pin Lee
    • Food Science and Preservation
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    • v.30 no.1
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    • pp.146-154
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    • 2023
  • To develop a multi-functional ingredient, the bioconversion of katsuobushi protein was optimized using Bacillus subtilis HA and Lactobacillus plantarum KS2020. The Dendropanax morbiferus extract (DME) culture with protease activity (102 unit/mL) was prepared by B. subtilis with 2% glucose and 1% skim milk through one day of alkaline fermentation. Katsuobushi protein was effectively hydrolyzed by the DME culture at 60℃ for 3 hours, resulting in a tyrosine content of 156.85 mg%. Subsequently, a second lactic acid fermentation was carried out with 10% monosodium glutamate (MSG) using L. plantarum KS2020 to produce higher levels of GABA. Following co-cultivation for three days, DME exhibited a pH of 8.3 (0% acidity). After seven days, the viable cell count of L. plantarum increased to 9.33 CFU/mL, but viable Bacillus cells were not detected. Taken together, a multi-functional ingredient with enriched GABA, peptides, probiotics, and umami flavor was developed through lactic acid fermentation using hydrolyzed katsuobushi protein. These results indicate that katsuobushi protein could be used as a byproduct to produce a palatable protein hydrolysate using alkaline-fermented DME culture as a proteolytic enzyme source.

Characteristics and anti-obesity effect of fermented products of coffee wine (커피발효물의 발효특성 및 항비만 효과)

  • So Hyun Park;Hyeon Hwa Oh;Do Youn Jeong;Young-Soo Kim
    • Food Science and Preservation
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    • v.30 no.4
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    • pp.703-715
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    • 2023
  • This study was conducted to investigate the fermentation characteristics and anti-obesity effects of acetic acid fermentation products of coffee wine. The live cell counts, soluble solids, pH and total acidity of the acetic acid unfermented coffee wine (AUFCW; day 0, before fermentation) were 6.35 log CFU/mL, 8.10 °Brix, 3.88, and 1.29%, respectively, while the acetic acid fermented coffee wine (AFCW; day 15, after fermentation) were 4.40 log CFU/mL, 8.57 °Brix, 3.07, and 7.45%, respectively. Pancreatic lipase inhibitory activity tended to increase as the acetic acid fermentation period increased. The anti-obesity effects of AFCW on 3T3-L1 cells, which was induced by MDI, were evaluated based on the lipid accumulation rate, leptin expression, and fat production-related gene expression (PPAR-γ and SREBP-1c) at the mRNA level. In the case of AFCW, the lipid accumulation rate and leptin expression were decreased to 69.37% and 50.20% at a concentration of 200 ㎍/mL, respectively, and the expression levels of PPAR-γ and SREBP-1c at the mRNA level were decreased to 79.89% and 48.81%, respectively. These results indicate that anti-obesity effect of acetic acid fermentation products could be increased by acetic acid fermentation of coffee wine.

The protective effect of Eucommia ulmoides leaves on high glucose-induced oxidative stress in HT-29 intestinal epithelial cells (고당으로 유도된 산화적 스트레스에 대한 두충 잎 추출물의 장 상피 세포 보호 효과)

  • Han Su Lee;Jong Min Kim;Hyo Lim Lee;Min Ji Go;Ju Hui Kim;Hyun Ji Eo;Chul-Woo Kim;Ho Jin Heo
    • Food Science and Preservation
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    • v.31 no.1
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    • pp.183-196
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    • 2024
  • This study investigated the protective effect of the aqueous extract of Eucommia ulmoides leaves (AEEL) against high glucose-induced human colon epithelial HT-29 cells. The 2,2'-azino-bis (3-ethyl benzothiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazy (DPPH) radical scavenging activities, ferric reducing/antioxidant power (FRAP), and malondialdehyde (MDA) analyses indicated that AEEL had significant antioxidant activities. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that AEEL increased cell viability against high glucose-, H2O2-, and lipopolysaccharide (LPS)-induced cytotoxicity in HT-29 cells. Also, the 2'-7'-dichlorodihydrofluorescein diacetate (DCF-DA) assay indicated that AEEL decreased intracellular reactive oxygen species (ROS) against high glucose-, H2O2-, and lipopolysaccharide (LPS)-induced cytotoxicity in HT-29 cells. AEEL showed inhibitory activities against α-glucosidase and inhibited the formation of advanced glycation end products (AGEs). AEEL showed significant positive effects on the viability and titratable acidity of L. brevis. The high-performance liquid chromatogram (HPLC) analysis identified chlorogenic acid and rutin as the major compounds of AEEL. These results suggested that AEEL has the potential to be used as a functional food source to suppress blood glucose levels and protect the gut from high glucose-induced oxidative stress.