Ultrasensitive Optical Detection of Reactive Oxygen Species Using Plasmonic-graphene Hybrid Interface

Hakchun KIM,Hyun Ji AN,Junhee PARK,Yohan LEE,Seungki LEE,Ji Hyeon OH,Nam Dong KIM,Jihwan SONG,Inhee CHOI 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10

Reactive oxygen species (ROS) produced as a by-product of mitochondrial respiration is defined as a molecule with strong oxidizing power and high reactivity. Generated-ROS is directly involved in the physiological regulation of cellular signaling pathways and oxidative stress. Thus, it is important to sensitively detect cellular ROS level of because ROS levels in cells plays an important role in determining the lifespan of cells. Herein, we present an ultrasensitive optical detection method of ROS, which is enhanced by tunable optical properties of plasmonic-graphene hybrid interface. ROS levels are monitored via plasmon resonance energy transfer (PRET) of the interaction between single plasmonic nanoprobes with redox-active cytochrome c. Surprisingly, the PRET signal is enhanced by transferring graphene layers onto the silver nanoparticle. Based on the PRET signals, the plasmonic-graphene hybrid interface exhibits a femtomolar level sensitivity for hydrogen peroxide and in-situ monitoring of ROS generated from the cells. We expect that the proposed ROS detection method using a plasmonic-graphene hybrid interface will be useful for investigating ROS level-dependent cellular abnormalities.

Beneficial Physiological Actions of Reactive Oxygen Species (ROS)

Jae-Myung Yoo,Mei Chen Jin,Dai-Eun Sok 충남대학교 약학대학 의약품개발연구소 2013 藥學論文集 Vol.28 No.-

Although reactive oxygen species (ROS) at high concentrations are known to cause non-specific damage to proteins, lipids and nucleic acids, a low level of ROS is known to exert beneficial effects through regulation of cellular signaling process or play a role as substrates for some physiologically important enzymes. Generally, such a biological function of ROS may be determined by the amount, period and localization of ROS. The former role may be implicated in redox regulation of some important proteins such as signal proteins, ion channels, and transcription factors. For the latter, ROS may be necessary for biosynthetic processes, including production of thyroid hormones and cross-linking of some proteins in extracellular systems. Concerning the formation of ROS, there may be multiple sources of ROS, including mitochondrial electron flow system, NADPH oxidases, xanthine oxidase, etc.. In contrast, there are ROS-degrading systems such as superoxide dismutase, catalase, peroxidase, protein thiols etc.. Therefore, the balance between ROS formation and degradation may be important for physiological roles of ROS. In this respect, antioxidant supplementation is to be carefully implemented because antioxidants may non-specifically limit the beneficial role of ROS. For this purpose, a specific inhibition of ROS-producing enzymes may be an approach promising clinical efficacy.

Reactive oxygen species (ROS) activity of ambient fine particles (PM_2.5) measured in Seoul, Korea

Park, Jieun,Park, Eun Ha,Schauer, James J.,Yi, Seung-Muk,Heo, Jongbae Elsevier 2018 Environment international Vol.117 No.-

<P><B>Abstract</B></P> <P>Substantial increase in level of particulate matter has raised concerns in South Korea recently. Ambient particulate matter is classified as Group I carcinogen (IARC, 2013) and multiple epidemiological studies has demonstrated adverse health effects due to exposure of particulate matter. Fine particulate matter (PM<SUB>2.5</SUB>) which has a diameter <2.5 μm is likely to penetrate deeply into lung and is known to be eliciting adverse health effects. A number of epidemiological studies have been conducted on adverse health effects of PM-related diseases and mortality rate, yet particulate matter (PM)-induced reactive oxygen species (ROS) activity at the cellular level has not been actively studied in Korea. This study assessed PM-induced oxidative potential by exposure of collected ambient PM<SUB>2.5</SUB> samples to the rat alveolar macrophage cell line. The characteristics of PM<SUB>2.5</SUB> in Korea were further characterized by linking chemical constituents and contributing sources to ROS. PM<SUB>2.5</SUB> mass concentration during the cold season was relatively higher than mass concentration during the warm season and chemical constituents except for Secondary Organic Carbon (SOC) and SO<SUB>4</SUB> <SUP>2−</SUP> which both showed similar trends in both the cold and cold seasons. The concentration of crustal elements was especially high during the cold season which can be an indication of long range transport of Asian dust. Water soluble organic carbon and water soluble transition metals (Cr and Zn) were also shown to be correlated to oxidative potential and metals such as As and V were shown to have a high contribution to ROS activity according to stepwise multiple linear regression. Principal Component Analysis (PCA) results identified six factors that can be interpreted as soil, mobile, industry, secondary inorganic aerosol, secondary organic aerosol and oil combustion. Moreover, through Principal Component Regression (PCR), industry, soil, mobile and SIA were shown to be statistically significant sources in a relation to ROS activity.</P> <P><B>Highlights</B></P> <P> <UL> <LI> PM-induced oxidative potential was assessed through cellular responses. </LI> <LI> WSOC and meals (As and V) exhibited high positive association with the ROS activity. </LI> <LI> Industry and soil sources were strongly correlated with ROS activity. </LI> </UL> </P>

Effect of Vitamin E Against the Cytotoxicity of Reactive Oxygen Species on Vascular Endothelial Cells

Kwon O-Yu,Park Seung-Taeck 대한의생명과학회 2006 Biomedical Science Letters Vol.12 No.3

Reactive oxygen species (ROS) is one of the main pathological factors in endothelial disorder. For example, an atherosclerosis is induced by the dysfunction of vascular endothelial cells. The dysfunction of vascular endothelial cells cascades to secrete intercellular adhesion molecule (ICAM)-l substance by ROS. Therefore, The ROS is regraded as an important factor of the injury of vascular endothelial cells and inducement of atherosclerosis. Oxygen radical scavengers playa key role to prevention of many diseases mediated by oxidative stress of ROS. In this study, the toxic effect of ROS on vascular endothelial cells and the effect of antioxidant, vitamin E on bovine pulmonary vascular endothelial cell line (BPVEC) treated with hydrogen peroxide were examined by the colorimetric assay. ROS decreased remarkably cell viability according to the dose- and time-dependent manners. In protective effect of vitamin E on BPVEC treated with hydrogen peroxide, vitamin E increased remarkably cell viability compared with control after BPVEC were treated with $15{\mu}M$ hydrogen peroxide for 6 hours. From these results, it is suggested that ROS has cytotoxicity on cultured BPVEC and oxygen radical scavenger such as vitamin E is very effective in prevention of oxidative stress-induced cytotoxicity.

Crosstalk between reactive oxygen species and pro-inflammatory markers in developing various chronic diseases: a review

Yazan Ranneh,Faisal Ali,Abdah Md Akim,Hasiah Abd. Hamid,Huzwah Khazaai,Abdulmannan Fadel 한국응용생명화학회 2017 Applied Biological Chemistry (Appl Biol Chem) Vol.60 No.3

The inflammation process in the human body plays a central role in the pathogenesis of many chronic diseases. In addition, reactive oxygen species (ROS) exert potentially a decisive role in human body, particularly in physiological and pathological process. The chronic inflammation state could generate several types of diseases such as cancer, atherosclerosis, diabetes mellitus and arthritis, especially if it is concomitant with high levels of pro-inflammatory markers and ROS. The respiratory burst of inflammatory cells during inflammation increases the production and accumulation of ROS. However, ROS regulate various types of kinases and transcription factors such nuclear factor-kappa B which is related to the activation of pro-inflammatory genes. The exact crosstalk between pro-inflammatory markers and ROS in terms of pathogenesis and development of serious diseases is still ambitious. Many studies have been attempting to determine the mechanistic mutual relationship between ROS and proinflammatory markers. Therefore hereby, we review the hypothetical relationship between ROS and pro-inflammatory markers in which they have been proposed to initiate cancer, atherosclerosis, diabetes mellitus and arthritis.

북극 스발바드 군도 콩스피요르드에서 채집한 해조류 추출물의 항산화 활성

이정임,김유아,예성수,한태준,강성호,김지희,서영완,공창숙 한국해양과학기술원 2009 Ocean and Polar Research Vol.31 No.2

We examined the intracellular antioxidative effects of 20 Arctic seaweed extracts in Raw 264.7 cells. Each seaweed species was subjected to extraction using acetone/dichloromethane and methanol, respectively, after which the extracts were combined and used as the test sample. The antioxidant ability of all 20 seaweeds extracts was evaluated using four different activity tests, including the degree of occurrence of intracellular reactive oxygen species (ROS), ONOO−, and lipid peroxidation in Raw 264.7 cells, as well as the extent of oxidative damage of genomic DNA purified from Raw 264.7 cells. Crude extracts from Monostroma obscurum, Alaria esculnta, Laminaria digitata, Desmarestia aculeata, Chorda filum, Ptilota seriata, Phycidrys rubens, Devaleraea ramentacea and Palmaria palmata exhibited significant scavenging effects on the generation of intracellular ROS. Among them, Monostroma obscurum and Phycidrys rubens significantly inhibited membrane lipid peroxidation and DNA oxidation. Moreover, Phycidrys rubens exhibited scavenging effects on peroxynitrite generated from SIN-1. We examined the intracellular antioxidative effects of 20 Arctic seaweed extracts in Raw 264.7 cells. Each seaweed species was subjected to extraction using acetone/dichloromethane and methanol, respectively, after which the extracts were combined and used as the test sample. The antioxidant ability of all 20 seaweeds extracts was evaluated using four different activity tests, including the degree of occurrence of intracellular reactive oxygen species (ROS), ONOO−, and lipid peroxidation in Raw 264.7 cells, as well as the extent of oxidative damage of genomic DNA purified from Raw 264.7 cells. Crude extracts from Monostroma obscurum, Alaria esculnta, Laminaria digitata, Desmarestia aculeata, Chorda filum, Ptilota seriata, Phycidrys rubens, Devaleraea ramentacea and Palmaria palmata exhibited significant scavenging effects on the generation of intracellular ROS. Among them, Monostroma obscurum and Phycidrys rubens significantly inhibited membrane lipid peroxidation and DNA oxidation. Moreover, Phycidrys rubens exhibited scavenging effects on peroxynitrite generated from SIN-1.

In vivo imaging of reactive oxygen species (ROS)-producing pro-inflammatory macrophages in murine carotid atheromas using a CD44-targetable and ROS-responsive nanosensor

Eun Jin Park,Joon Woo Song,Hyun Jung Kim,Chang-Soo Kim,Yeong Jun Song,Dae Hyeok Yang,Hongki Yoo,Jin Won Kim,Kyeongsoon Park 한국공업화학회 2020 Journal of Industrial and Engineering Chemistry Vol.92 No.-

In this study, we report the in vivo imaging of reactive oxygen species (ROS)-overproducing pro-inflammatory macrophages in atherosclerotic plaques using afluorescent ROS nanosensor. We designedthe nanosensor by chemically conjugating hyaluronic acid (HA, a targeting ligand for CD44 receptor),chlorin e6 [Ce6; a near-infraredfluorescent (NIRF) dye], and a thioketal (TK) linker (ROS-degradablelinker). The self-assembled nanosensor emitted weak NIRF signals in normal physiological conditions,whereas it emitted strong NIRF signals under ROS-abundant conditions. The cytocompatible nanosensorshowed higher intracellular internalization via receptor-mediated endocytosis, which enabled thevisualization of intracellular ROS in pro-inflammatory macrophages. Moreover, we demonstrated thatthe nanosensor enabled the successful targeting and imaging of CD44- and ROS-overproducing pro-inflammatory macrophages in atherosclerotic plaques, as validated by confocal microscopy andimmunohistological analyses. These data suggest that our ROS nanosensor is suitable for ROS imaging inpro-inflammatory macrophages in vitro and in vivo and will be a promising approach for imagingatherosclerotic tissues and evaluating the effects of antioxidants.

U937 인체혈구암세포에서 diallyl trisulfide에 의한 mitotic arrest와 apoptosis 유발

박현수(Hyun Soo Park),이준혁(Jun Hyuk Lee),손병일(Byoung Yil Son),최병태(Byung Tae Choi),최영현(Yung Hyun Choi) 한국생명과학회 2013 생명과학회지 Vol.23 No.5

본 연구에서는 마늘에서 유래된 생리활성 물질인 diallyl trisulfide (DATS) 처리에 따른 U937 인체혈구암세포의 증식억제가 apoptosis 및 cell cycle arrest 유발과 관련이 있는지 조사하였다. U937 세포증식은 DATS에 의해 농도 및 시간 의존적으로 감소함을 확인 하였고, 이는 apoptosis에 의한 직접적인 세포죽음과 CDK1 및 cyclin B1의 발현 증가 및 histone H3의 인산화와 연관된 mitotic arrest와 관련이 있음을 알 수 있었다. 또한 DATS 처리 초기에 reactive oxygen species (ROS)의 생성이 매우 증가되었으나, ROS scavenger (N-acetyl-l-cysteine)에 의한 인위적 ROS 생성의 억제는 DATS에 의한 apoptosis 및 mitotic arrest를 완벽하게 차단시켰다. 이는 U937 세포에서 DATS에 의해 유도된 apoptosis 및 mitotic arrest가 ROS에 의해 매개된다는 것을 의미하며, 본 연구의 결과는 DATS가 인체혈구암세포에서 세포증식억제와 관련된 항암기전을 이해할 수 있는 기초자료로서 매우 유용하게 사용될 것이라 생각된다. Diallyl trisulfide (DATS), one of the major organosulfur components of garlic (Allium sativum), has various biological effects such as anti-microbial and anti-cancer activities. However, the molecular mechanisms of growth inhibition related to cell cycle arrest are poorly understood. In this study, we investigated the effects of DATS on cell cycle progression in U937 human leukemia cells. Treatment with DATS in U937 cells resulted in inhibition of cell viability through G2/M arrest and apoptosis. DATS-induced G2/M arrest was associated with up-regulation of cyclin B1 and cyclin-dependent kinase 1 (CDK1). DATS also significantly increased levels of phospho-histone H3, which is a mitosis-specific marker, indicating that DATS induced mitotic arrest but not G2 arrest in U937 cells. DATS treatment also generated the reactive oxygen species (ROS) in U937 cells; however, pretreatment with N-acetyl-l-cysteine (NAC), a ROS scavenger, significantly attenuated DATS-induced mitotic arrest and apoptosis. Taken together, our data indicate that DATS exhibits anti-cancer effects through mitotic arrest and apoptosis in a ROS-dependent manner.

AGS 인체 위암세포에서 황흑산에 의한 ROS 생성 및 caspase 활성 의존적 apoptosis 유발

홍수현(Su Hyun Hong),박철(Cheol Park),김경민(Kyoung Min Kim),최영현(Yung Hyun Choi) 한국생명과학회 2015 생명과학회지 Vol.25 No.11

황흑산은 동의보감의 복강과 장옹의 처방을 위해 기록된 처방전으로 오랫동안 사용되어 왔으나, 항암 효능에 대한 구체적인 연구는 전혀 이루어진 바 없다. 본 연구에서는 AGS 인체 위암세포를 대상으로 황흑산 처리에 의한 증식억제와 연관된 apoptosis 유발 및 관련 기전 연구를 수행하였다. AGS 위암세포에 황흑산 추출물을 처리함에 처리 농도 의존적으로 증식이 억제되었으며, 이는 apoptosis 유발과 연관성이 있음을 핵의 형태적 변형과 sub-G1기 세포의 축적 등으로 확인하였다. 황흑산 추출물에 의한 apoptosis 유도에는 pro-apoptotic Bax 단백질의 발현증가와 anti-apoptotic Bcl-2의 발현 감소 및 미토콘드리아에서 세포질로의 cytochrome c 유리와 연관성이 있었으며, 세포 내 활성산소종(reactive oxygen species, ROS)의 축적을 증가시켰다. 또한 황흑산 추출물에 의한 apoptosis 유발은 caspases (caspase-3, -8 및 -9)의 활성을 증가시켰으며, poly (ADP-ribose)-polymerase 단백질의 단편화를 초래하였다. 그러나 ROS scavenger 및 pan-caspases inhibitor는 황흑산 추출물에 의한 apoptosis의 유발을 거의 완벽하게 억제하였으며, 암세포의 증식억제도 차단하였다. 이상의 결과는 황흑산 추출물에 의한 apoptosis가 ROS 생성 및 caspase 활성 의존적으로 일어남을 의미하는 것으로 황흑산의 항암기전 해석을 이해하고 향후 지속적인 연구를 위한 유용한 자료로 사용될 것이다. Hwangheuk-san (HHS) is a Korean multi-herb formula comprising four medicinal herbs. HHS, which was recorded in “Dongeuibogam,” has been used to treat patients with inflammation syndromes and digestive tract cancer for hundreds of years. However, little is known about its anti-tumor efficacy. The present study investigated the pro-apoptotic effect and mode of action of HHS against AGS human gastric carcinoma cells. HHS inhibited the cell growth of AGS cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies, chromatin condensation, and an accumulation of cells in the sub-G1 phase. HHS-induced apoptotic cell death was associated with the up-regulation of pro-apoptotic Bax protein expression, down-regulation of antiapoptotic Bcl-2 protein, and the release of cytochrome c from mitochondria to the cytosol. The treatment of AGS cells with HHS significantly elevated the generation of reactive oxygen species (ROS). Additionally, apoptosis-inducing concentrations of HHS induced the activation of both caspase-9 and -8, initiator caspases of the mitochondrial-mediated intrinsic and death receptor-mediated extrinsic pathways, respectively, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose)-polymerase. However, ROS scavenger and pan-caspases inhibitor significantly blocked HHS-induced growth inhibition and apoptosis. Taken together, these findings suggest that HHS induces apoptosis through ROS- and caspase-dependent mechanisms and that HHS may be a potential chemotherapeutic agent for the control of human gastric cancer.

적작약 꽃 추출물의 활성산소 억제와 항염증 및 MMP-1 발현 억제능 효과에 관한 연구

이재남(Jae-Nam Lee),김영삼(Young-Sam Kim) 한국유화학회 2018 한국응용과학기술학회지 Vol.35 No.3

This study attempted to investigate the effects of Paeonia Lactiflora Pallas (P. lactiflora) on the inhibition of oxygen free radical, anti-inflammation and MMP-1 inhibitory activity and examine its possibility as a functional cosmetic material. For test methods, the inhibition of oxygen free radical after measuring reactive oxygen species (ROS) in the cell, cytotoxicity assessment and anti-inflammation were measured, and MMP-1 inhibitory effects in the HDF cell were measured. According to the test, the inhibition of ROS was confirmed in RAW 264.7 and HDF cells. In terms of cytotoxicity assessment, 90% or higher cell viability was detected at 5/10μg/mL Paeonia Lactiflora Pallas extract while it was 80% or higher at other concentration levels in both RAW 264.7 and HDF cells. In addition, NO production was inhibited in the RAW 264.7 cell while MMP-1 was significantly inhibited in the HDF cell. The above results reveal a possibility of Paeonia Lactiflora Pallas extract as a functional cosmetic material after confirming the inhibition of ROS synthesis in the cell, antioxidant and anti-inflammatory effects by inhibiting NO synthesis, low toxicity on skin cells and anti-aging effect through MMP-1 inhibition. 본 연구는 적작약 꽃 추출물의 활성산소 억제와 항염증 및 MMP-1 발현 억제능 효과에 관해 알아보고 기능성 화장품 소재로써의 가능성을 확인하고자 하였다. 본 실험 방법으로는 세포 내 ROS 측정을 통한 활성산소 억제효과와 세포 독성 평가 및 항염증 측정, HDF 세포에서의 MMP-1의 발현 억제능 효과를 측정하고자 하였다. 실험 결과, RAW 264.7 세포와 HDF 세포 내에서 ROS로 억제효과를 확인하 였고, 세포 독성평가는 적작약 꽃 추출물 5, 10 μg/mL 처리 농도에서 90% 이상의 세포 생존율, 그 외 처리 농도에서는 80% 이상의 세포 생존율을 확인하였다. 또한 RAW 264.7 세포에서의 NO 생성 억제와 HDF 세포에서 MMP-1의 발현 억제능이 유의하게 억제되는 것을 확인하였다. 이상의 결과를 종합하면, 적작약 꽃 추출물의 세포 내 ROS 생성억제와 NO 생성 억제로 항산화와 항염증 효과, 피부세포에 대한 낮은 독성, MMP-1의 발현 억제를 통한 노화 효과가 확인됨에 따라 기능성 화장품 소재로써의 활용 가능성을 확인할 수 있었다.