• Journal of Ginseng Research
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• 제37권4호
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• pp.413-424
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• 2013

Korean Red Ginseng extract (KRGE) is a traditional herbal medicine utilized to prevent endothelium dysfunction in the cardiovascular system; however, its underlying mechanism has not been clearly elucidated. We here examined the pharmacological effect and molecular mechanism of KRGE on apoptosis of human umbilical vein endothelial cells (HUVECs) in a serum-deprived apoptosis model. KRGE protected HUVECs from serum-deprived apoptosis by inhibiting mitochondrial cytochrome c release and caspase-9/-3 activation. This protective effect was significantly higher than that of American ginseng extract. KRGE treatment increased antiapoptotic Bcl-2 and Bcl-$X_L$ protein expression and Akt-dependent Bad phosphorylation. Moreover, KRGE prevented serum deprivation-induced subcellular redistribution of these proteins between the mitochondrion and the cytosol, resulting in suppression of mitochondrial cytochrome c release. In addition, KRGE increased nitric oxide (NO) production via Akt-dependent activation of endothelial NO synthase (eNOS), as well as inhibited caspase-9/-3 activities. These increases were reversed by co-treatment of cells with inhibitors of eNOS and phosphoinositide 3-kinase (PI3K) and pre-incubation of cell lysates in dithiothreitol, indicating KRGE induces NO-mediated caspase modification. Indeed, KRGE inhibited caspase-3 activity via S-nitrosylation. These findings suggest that KRGE prevents serum deprivation-induced HUVEC apoptosis via increased Bcl-2 and Bcl-$X_L$ protein expression, PI3K/Akt-dependent Bad phosphorylation, and eNOS/NO-mediated S-nitrosylation of caspases. The cytoprotective property of KRGE may be valuable for developing new pharmaceutical means that limit endothelial cell death induced during the pathogenesis of vascular diseases.

• Journal of Ginseng Research
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• 제45권1호
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• pp.119-125
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• 2021

Background: Korean Red Ginseng (KRG) is a natural product with antiinflammatory and anticarcinogenic effects. We have previously reported that the endocrine-disrupting compound bisphenol A (BPA)-induced cyclooxygenase-2 (COX-2) via nuclear translocation of nuclear factor-kappa B (NF-κB) and activation of mitogen-activated protein kinase and promoted the migration of A549. Here, in this study, we assessed the protective effect of KRG on the BPA-induced reactive oxygen species (ROS) and expression of COX-2 and matrix metalloproteinase-9 (MMP-9) in A549 cells. Methods: The effects of KRG on the upregulation of ROS production and COX-2 and MMP-9 expression by BPA were evaluated by fluorescence-activated cell sorting (FACs) analysis, quantitative reverse transcription polymerase chain reaction, and western blotting. Antimigration ability by KRG was evaluated by migration assay in A549 cells. Results: KRG significantly suppressed the BPA-induced COX-2, the activity of NF-κB, the production of ROS, and the migration of A549 cells. These effects led to the downregulation of the expression of MMP-9. Conclusions: Overall, our results suggest that KRG exerts an antiinflammatory effect on BPA-treated A549 cells via the suppression of ROS and downregulation of NF-κB activation and COX-2 expression which leads to a decrease in cellular migration and MMP-9 expression. These results provide a new possible therapeutic application of KRG to protect BPA-induced possible inflammatory disorders.

• 대한약침학회지
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• 제24권1호
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• pp.24-31
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• 2021

Objectives: Accumulation of cellular reactive oxygen species (ROS) leads to oxidative stress. Increased production of ROS, such as superoxide anion, or a deficiency in their clearance by antioxidant defences, mediates cellular pathology. Grewia Spp fruits are a source of bioactive compounds and have notable antioxidant activity. Although the antioxidant capacity of Grewia Spp has been studied, there is very limited evidence that links the antioxidant activities of Grewia bicolor and Grewia flava to the inhibition of free radical formation associated with damage in biological systems. Methods: This study evaluated the protective effects of Grewia bicolor and Grewia flava extracts against free radical-induced oxidative stress and the resulting cytotoxicity effect using HeLa cells. Antioxidant properties determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and total phenolic content (TPC) assays showed significantly higher (p < 0.05) antioxidant activity in Grewia flava (ethanol extract) than Grewia flava (water extract) and Grewia bicolor (ethanol and water extracts). Results: Using 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide or MTT assay, cytotoxicity results showed that extracts of Grewia bicolor and Grewia flava were less toxic to HeLa cells at tested concentrations compared to the untreated control. This confirmed the low toxicity of these edible fruits at the tested concentrations in HeLa cells. Furthermore, hydrogen peroxide (H2O2)-induced cell loss was effectively reduced by pre-incubating HeLa cells with Grewia bicolor and Grewia flava extracts, with Grewia flava (ethanol extract) revealing better protection. Conclusion: The effect was speculated to be associated with the higher antioxidant activity of Grewia flava (ethanol extract). Additional studies will warrant confirmation of the mechanism of action of such effects.

• Journal of Ginseng Research
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• 제48권4호
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• pp.405-416
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• 2024

Background: Hypoxic pulmonary hypertension (HPH) is the main pathological change in vascular remodeling, a complex cardiopulmonary disease caused by hypoxia. Some research results have shown that ginsenoside Rg1 (Rg1) can improve vascular remodeling, but the effect and mechanism of Rg1 on hypoxia-induced pulmonary hypertension are not clear. The purpose of this study was to discuss the potential mechanism of action of Rg1 on HPH. Methods: C57BL/6 mice, calpain-1 knockout mice and Pulmonary artery smooth muscle cells (PASMCs) were exposed to a low oxygen environment with or without different treatments. The effect of Rg1 and calpain-1 silencing on inflammation, fibrosis, proliferation and the protein expression levels of calpain-1, STAT3 and p-STAT3 were determined at the animal and cellular levels. Results: At the mouse and cellular levels, hypoxia promotes inflammation, fibrosis, and cell proliferation, and the expression of calpain-1 and p-STAT3 is also increased. Ginsenoside Rg1 administration and calpain-1 knockdown, MDL-28170, and HY-13818 treatment showed protective effects on hypoxia-induced inflammation, fibrosis, and cell proliferation, which may be associated with the downregulation of calpain-1 and p-STAT3 expression in mice and cells. In addition, overexpression of calpain 1 increased p-STAT3 expression, accelerating the onset of inflammation, fibrosis and cell proliferation in hypoxic PASMCs. Conclusion: Ginsenoside Rg1 may ameliorate hypoxia-induced pulmonary vascular remodeling by suppressing the calpain-1/STAT3 signaling pathway.

• 대한화장품학회지
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• 제38권1호
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• pp.15-31
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• 2012

이전 연구에서 저자들은 여뀌 추출물의 항산화, 항노화, 항균 활성 및 여뀌 추출물을 함유한 크림의 보습 효과에 대한 결과를 보고한 바 있다. 본 연구에서는 여뀌 추출물과 그 주요 성분인 isoquercitrin의 사람 적혈구와 HaCaT 세포에서의 세포 보호 효과를 측정하였다. 여뀌 추출물의 피부 전달시스템으로 에토좀 및 탄성 리포좀을 제조하여 입자크기, 포집효율, 안정성 및 피부 흡수 증진 능력을 평가하였다. 적혈구 광용혈에서 여뀌 추출물과 isoquercitrin은 $5{\mu}g/mL$의 농도에서 ${\alpha}$-토코페롤보다도 큰 세포보호효과를 나타내었다. 여뀌 추출물은 HaCaT 세포에 대해 $50{\mu}g/mL$의 농도에서 독성을 나타내지 않았다, UVB $400mJ/cm^2$를 HaCaT cell에 조사하였을 때, 여뀌 추출물은 농도 의존적($12.5{\sim}50{\mu}g/mL$)으로 자외선으로부터 세포를 보호하였고, $25{\mu}g/mL$의 농도에서 양성 대조군(세포 생존율, 36 %)에 비하여 큰세포보호 활성(생존율, 90 %)을 나타내었다. 0.04 % 여뀌 추출물을 담지한 에토좀의 입자 크기는 173.0 nm, 포집효율은55.58 %이였다. 에토좀 제조 후 일주일동안 안정한 단분산 형태를 나타내었다. 피부 투과 실험 결과 에토좀은 일반 리포좀이나 에탄올 용액에서보다도 더 큰 피부 투과능을 보여주었다. 0.1 % 여뀌 추출물을 담지한 탄성 리포좀의 최적의 제형은 인지질 대 계면활성제($Tego^{(R)}$care 450)의 비율이 95 : 5으로 확인되었다. 0.1 % 여뀌 추출물 함유한 최적의 탄성 리포좀의 입자크기는 176.5 nm, 가변형성은 16.4, 포집효율은 68.8 %이었다. 여뀌 추출물을 담지한 탄성 리포좀은 계면활성제가 포함되지 않은 제형보다 더 큰 피부 투과능을 나타내었다.

• 대한화장품학회지
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• 제39권4호
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• pp.259-269
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• 2013

본 연구에서는 한국 및 중국산 감초(Glycyrrhiza uralensis), 그리고 우즈베키스탄산 감초(Glycyrrhiza glabra)를 대상으로 추출용매, 추출온도, 추출시간 등 추출조건별 추출물을 제조하고 이들 추출물들의 추출 수율과 항산화 활성을 비교하여 최적의 추출조건을 선정하였다. 항산화 활성 중 자유 라디칼(1,1-phenyl-2-picrylhydrazyl, DPPH) 소거활성은 85% 에탄올로 $60^{\circ}C$에서 6 h 동안 추출한 조건에서 한국 감초로부터 얻은 추출물이 가장 높은 활성(46.05%)을 나타내었다. 루미놀 발광법을 이용한 추출물들의 총 항산화능 측정 실험과 피부 광노화에 중요한 $^1O_2$으로 유도된 세포막 손상에 대한 추출물들의 세포 보호 효과를 측정한 실험 모두에서도 위의 조건에서 가장 높은 항산화 활성을 나타내었다. 특히, 한국 감초는 ${\tau}_{50}$이 116.4 min으로 비교 물질인 (+)-${\alpha}$-tocopherol (28.5 min)보다 약 4 배나 높은 세포 보호 효과를 나타내었으며, 추출 수율은 18.75%로 우즈베키스탄 및 중국 감초보다 각각 1.2 배 및 2.5 배의 추출 수율을 나타내었다. 따라서, 본 연구 결과는 항산화 소재로 화장품에 응용하기 위하여 감초로부터 추출물을 얻기 위한 최적의 조건은 85% 에탄올로 $60^{\circ}C$에서 6시간 동안 추출하는 것임을 보여주었다.

• Journal of Ginseng Research
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• 제41권2호
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• pp.169-179
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• 2017

Background: Ginsenosides are the main pharmacological components of Panax ginseng root, which are thought to be primarily responsible for the suppressing effect on oxidative stress. Methods: 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and oxygen radical absorption capacity were applied to evaluate the antioxidant activities of the ginsenosides. Human embryonic kidney 293 (HEK-293) cells were incubated with ginsenosides extracted by pulsed electric field (PEF) and solvent cold soak extraction (SCSE) for 24 h and then the injury was induced by $40{\mu}M$ $H_2O_2$. The cell viability and surface morphology of HEK-293 cells were studied using MTS assay and scanning electron microscopy, respectively. Dichloro-dihydro-fluorescein diacetate fluorescent probe assay was used to measure the level of intracellular reactive oxygen species. The intracellular antioxidant activities of ginsenosides were evaluated by cellular antioxidant activity assay in HepG2 cells. Results: The PEF extracts displayed the higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and stronger oxygen radical absorption capacity (with an oxygen radical absorption capacity value of $14.48{\pm}4.04{\mu}M\;TE\;per\;{\mu}g/mL$). The HEK-293 cell model also suggested that the protective effect of PEF extracts was dose-dependently greater than SCSE extracts. Dichloro-dihydro-fluorescein diacetate assay further proved that PEF extracts are more active (8% higher than SCSE extracts) in reducing intracellular reactive oxygen species accumulation. In addition, scanning electron microscopy images showed that the HEK-293 cells, which were treated with PEF extracts, maintained more intact surface morphology. Cellular antioxidant activity values indicated that ginsenosides extracted by PEF had stronger cellular antioxidant activity than SCSE ginsenosides extracts. Conclusion: The present study demonstrated the antioxidative effect of ginsenosides extracted by PEF in vitro. Furthermore, rather than SCSE, PEF may be more useful as an alternative extraction technique for the extraction of ginsenosides with enhanced antioxidant activity.

• Journal of Dental Anesthesia and Pain Medicine
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• 제17권1호
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• pp.21-28
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• 2017

Background: The skin consists of tightly connected keratinocytes, and prevents extensive water loss while simultaneously protecting against the entry of microbial pathogens. Excessive cellular levels of reactive oxygen species can induce cell apoptosis and also damage skin integrity. Propofol (2,6-diisopropylphenol) has antioxidant properties. In this study, we investigated how propofol influences intracellular autophagy and apoptotic cell death induced by oxidative stress in human keratinocytes. Method: The following groups were used for experimentation: control, cells were incubated under normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$) without propofol; hydrogen peroxide ($H_2O_2$), cells were exposed to $H_2O_2$ ($300{\mu}M$) for 2 h; propofol preconditioning (PPC)/$H_2O_2$, cells pretreated with propofol ($100{\mu}M$) for 2 h were exposed to $H_2O_2$; and 3-methyladenine $(3-MA)/PPC/H_2O_2$, cells pretreated with 3-MA (1 mM) for 1 h and propofol were exposed to $H_2O_2$. Cell viability, apoptosis, and migration capability were evaluated. Relation to autophagy was detected by western blot analysis. Results: Cell viability decreased significantly in the $H_2O_2$ group compared to that in the control group and was improved by propofol preconditioning. Propofol preconditioning effectively decreased $H_2O_2$-induced cell apoptosis and increased cell migration. However, pretreatment with 3-MA inhibited the protective effect of propofol on cell apoptosis. Autophagy was activated in the $PPC/H_2O_2$ group compared to that in the $H_2O_2$ group as demonstrated by western blot analysis and autophagosome staining. Conclusion: The results suggest that propofol preconditioning induces an endogenous cellular protective effect in human keratinocytes against oxidative stress through the activation of signaling pathways related to autophagy.

• 대한화장품학회지
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• 제48권2호
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• pp.157-167
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• 2022

본 연구에서는 7 개의 아미노산으로 이루어진 헵타펩타이드의 섬유아세포 활성 증가 및 광노화 조건에서의 세포 손상 억제 효과를 확인하였다. 실험 결과 헵타펩타이드 처리 시 섬유아세포 증식 및 세포외기질(extracellular matrix, ECM) 구성 인자의 발현이 증가되었다. 그리고 자외선 A (ultraviolet A, UVA) 조사에 의해 유도된 광노화조건에서 감소된 세포 생존율이 헵타펩타이드에 의해 증가되었고, UVA 조사에 의해 유도된 세포 사멸, 기질금속단백질분해효소-1(matrix metalloproteinases-1, MMP-1) 발현 및 세포 내 활성산소종(reactive oxygen species, ROS) 수준이 헵타펩타이드에 의해 감소되었다. UVA 조사 시 나타나는 transforming growth factor-β (TGF-β)/smad 기전 억제와 그에 따른 ECM 구성 인자 발현 감소 또한 헵타펩타이드에 의해 회복되었다. 또 다른 광노화 유도 조건으로 heat shock을 주었고 헵타펩타이드를 전 처리 하였을 때 heat shock에 의한 mitogen-activated protein kinase (MAPK) 인산화 및 MMP-1 발현이 억제됨을 확인할 수 있었다. 이 결과를 종합해 볼 때, 본 연구의 헵타펩타이드는 섬유아세포의 활성을 촉진하며, 광노화 유도 모델로 사용된 UVA 조사 및 heat shock 조건에서도 세포 내 ROS 억제 효과를 보여 세포 손상에 대한 회복 및 보호 효과를 나타내는 것으로 보인다. 이러한 진피 보호 효과를 갖는 헵타펩타이드는 향 후 신규 화장품 소재로 응용될 수 있을 것으로 기대된다.

• 대한치과마취과학회지
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• 제14권1호
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• pp.41-47
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• 2014

Background: Autophagy is a self-eating process that is important for balancing sources of energy at critical times in development and in response stress. Autophagy also plays a protective role in removing clearing damaged intracellular organelles and aggregated proteins as well as eliminating intracellular pathogens. The purpose of the present study was to examine the protective effect of propofol against hypoxic damage using keratinocytes. Methods: Human keratinocytes (HaCaT cells) were obtained from the American Type Culture Collection. Propofol which were made by dissolving them in DMSO were kept frozen at $-4^{\circ}C$ until use. The stock was diluted to their concentration with DMEM when needed. Prior to propofol treatment cells were grown to about 80% confluence and then exposed to propofol at different concentrations (0, 25, 50, 75, $100{\mu}M$) for 2 h pretreatment. Cell viability was measured using a quantitative colorimetric assay with thiazolyl blue tetrazolium bromide (MTT assay), and fluorescence microscopy and western blot analysis were used for evaluation of autophagy processes. Results: The viability of propofol-treated HaCaT cells was increased in a dose-dependent manner. Propofol did not show any significant toxic effect on the HaCaT cells. The autophagy inhibitor, 3-methyladenine, reduced cell viability of hypoxia-injured HaCat cells. Fluorescence microscopy and western blot analysis showed propofol induce autophagy pathway signals. Conclusions: Propofol enhanced viability of hypoxia-injured HaCaT cells and we suggest propofol has cellular protective effects by autophagy signal pathway activation.