• Natural Product Sciences
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• 제21권2호
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• pp.128-133
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• 2015

Lindera obtusiloba has been widely used as a traditional medicine for the treatment of lots of diseases, including abdominal pain, bruise, and hepatocirrhosis. Here in this study, we elucidated the lifespan-extending effect of methanolic extract of Lindera obtusiloba (MLO) using Caenorhabditis elegans model system. We found that MLO has potent lifespan extension activities under normal culture condition. Then, we determined the protective effects of MLO on the stress conditions such as osmotic, thermal and oxidative stress. To reveal possible mechanism of MLO-mediated lifespan, we further investigated the effect of MLO on the antioxidant enzyme activities and intracellular ROS levels. Our results demonstrated that superoxide dismutase and catalase activities were significantly up-regulated by MLO treatment, resulted in reduced intracellular ROS levels. In this work, we also tested whether MLO-mediated longevity activity was associated with aging-related factors such as food intake and growth. Our data revealed that both of pharyngeal pumping rate and body length were significantly shifted by MLO treatment, indicating these factors were involved in MLO's lifespan-extension effects. Although MLO induces reduction in food intake, the body movement of MLO-fed aged worms was not decreased, compared to untreated control worms, indicating MLO might extend lifespan without affecting healthspan.

• 생약학회지
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• 제49권1호
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• pp.40-46
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• 2018

Chaenomeles sinensis (Thouin) Koehne fruit (Rosaceae) has been used as a traditional medicine in Korea, Japan and China to treat sore throat, diarrhea and inflammation. The ethanol extract of C. sinensis fruit was successively partitioned as methylene chloride, ethyl acetate, n-butanol and $H_2O$ soluble fractions. Among those fractions, the n-butanol fraction showed the most potent DPPH radical scavenging and superoxide quenching activities. To verify antioxidant activities, the n-butanol fraction was checked the activities of superoxide dismutase (SOD) and catalase activities, and intracellular ROS levels and oxidative stress tolerance in Caenorhabditis elegans. Furthermore, to see if increased stress tolerance of worms by treating of the n-butanol fraction was due to regulation of stress-response gene, we quantified SOD-3 expression using transgenic strain. Consequently, the n-butanol fraction elevated SOD and catalase activities of C. elegans, and reduced intracellular ROS accumulation in a dose-dependent manner. Moreover, the n-butanol fraction-treated CF1553 worms exhibited significantly higher SOD-3::GFP intensity.

• 한국식품과학회지
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• 제49권2호
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• pp.222-227
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• 2017

본 연구에서는 금 나노입자 용액을 생장하는 6년근 인삼에 직접 시비해서 금 나노입자가 전이된 황금인삼을 열수 추출하여 총페놀 함량, 총플라보노이드 함량, 산화방지능 및 신경세포 보호능을 평가하였다. $1^{\circ}Bx$의 황금홍삼 추출물은 총페놀과 총플라보노이드 함량이 각각 212.2 mg GAE와 3.5 mg CE였다. ABTS, DPPH 및 ORAC 법으로 측정시, 황금홍삼 추출물의 산화방지능은 각각 272.3, 141.2, $868.4mg\;VCE/^{\circ}Bx$였다. 황금홍삼 추출물은 과산화수소로부터 유래한 세포 내 산화스트레스를 감소시켜 PC-12 신경세포의 생존율을 농도 의존적으로 증가시켰다. 또한, 황금홍삼 추출물은 신경전달물질인 아세틸콜린을 가수분해하는 AChE 및 BChE 활성을 억제하였다. 이러한 결과는 금 나노입자를 처리한 홍삼을 이용한 산화방지 및 신경손상억제 소재로 활용할 가능성을 제시하였다.

• 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.

• 한국식품영양학회지
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• 제25권1호
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• pp.123-131
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• 2012

The objective of the present study was to evaluate the potential antidiabetic and antioxidant effect of the ethanol extract from Chrysanthemum cornarium L. var. spatiosum(CSE) against alloxan-induced oxidative stress in pancreatic ${\beta}$-cells, HIT-T15. In this study, the antidiabetic effect of CSE was examined using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliu bromide(MTT) cell proliferation assay, lactate dehydrogenase(LDH) release assay, $NAD^+$/NADH ratio and insulin secretion. To further investigate whether CSE is involved in the antioxidant activity of alloxan-damaged HIT-T15 cells, its antioxidant effect against alloxan-induced oxidative stress was measured in HIT-T15 cells by determining the levels of antioxidant enzymes including superoxide dismutase(SOD), glutathione S-transferase(GST), glutathione reductase(GR) and glutathione peroxidase(GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered $NAD^+$/NADH ratio and insulin secretion in HIT-T15 cells. However, CSE significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular NAD+/NADH ratio and insulin secretion were also significantly increased by 1.7-fold and 1.3-fold, respectively, after treatment with 100 ${\mu}g/m{\ell}$ CSE. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while CSE significantly elevated the levels of antioxidant enzymes. These findings suggest that CSE could have a protective effect against cytotoxicity and dysfunction of pancreatic cells in the presence of alloxan-induced oxidative stress.

• Tuberculosis and Respiratory Diseases
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• 제50권5호
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• pp.579-590
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• 2001

연구배경 : 노화란 반응성산소종에 의한 산화적 손상이 축적되어 세포의 기능의 저하로 초래된다고 주장되고 있다. 본 연구에서 1, 4, 8, 12개월된 정상 생쥐의 폐에서 반응성 산소종 생성의 초기에 중요한 방어기전인 superoxide dismutases(SODs) mRNA와 hydroxy radical($OH{\cdot}$)에 의한 산화적 손상의 마지막 방어기전인 metallothionein(MT) mRNA의 함량과 paraquat에 의한 유도능을 조사하였다. 방 법 : 산화적손상은 세포내에서 superoxide을 생성하는 paraquat를 투여한 후 SODs와 MT mRNAs을 RT-PCR 방법으로 측정하였다. 결 과 : 정상생쥐의 폐의 노화과정에서 Mn-SOD와 Cu/Zn-SOD, 그리고 MT mRNA의 항정상태 함량을 조사한 결과, Mn-SOD mRNA는 8개월까지는 증가하다가 12개월에서 감소하였으나, Cu/Zn-SOD mRNA는 별 변화가 없었고 12개월에 감소하였다. 그러나 MT mRNA는 어느 연령에도 변화가 없었다. 산화성 스트레스를 유발하는 paraquat에 의하여 Mn-SOD mRNA는 4개월 부터 유도되지 않았으며, Cu/Zn-SOD mRNA는 전혀 유도되지 않았다. MT mRNA는 관찰한 전 연령에서 유도되었다. 결 론 : 노화과정에서 SODs유도능의 이상이 노화의 원인 인자로 작용하고, 반면에 MT의 정상적인 유도능이 산화적 스트레스에 대하여 방어 역할을 할 가능성을 시사해주고 있다. 이와 같은 결과는 연령이 증가함에 따라 Mn-SOD, Cu/Zn-SOD, 및 MT의 발현과 유도능의 차이로 인하여 반응성 산소종에 의한 산화적 손상의 정도의 차이가 노화의 속도를 결정하는 하나의 인자로 작용할 가능성 이 있을 것으로 생각된다.

• 약학회지
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• 제49권2호
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• pp.156-161
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• 2005

Tetrahydropapaveroline (THP) at 5-15 ${\mu}$M has been found to induce L-DOPA-induced oxidative apoptosis in PC12 cells. In this study, the inhibitory effects of THP on dopamine bios ynthesis in PC12 cells and tyrosine hydroxylase (TH) activity in bovine adrenal were investigated. Treatment of PC12 cells with THP at 2.5-10 ${\mu}$M significantly decreased the intracellular dopamine content in a concentration-dependent manner (18.3% inhibition at 10 ${\mu}$M THP). In these conditions, TH activity was markedly inhibited by the treatment with THP at 2.5-10 ${\mu}$M in PC12 cells (23.4% inhibition at 10 $\mu$ M THP). In addition, THP had an inhibitory effect on bovine adrenal TH activity IC50 value, 153.9${\mu}$M). THP exhibited uncompetitive inhibition on bovine adrenal TH activity with a substrate L-tyrosine with the KI value of 0.30 mM. Treatment with L-DOPA at 20~50 ${\mu}$M increased the intracellular dopamine content in PC12 cells, and the increase in dopamine content by L-DOPA was inhibited in part when THP at non-cytotoxic (5-10 ${\mu}$M) or cytotoxic (15${\mu}$M) concentrations was associated with L-DOPA (20 and 50 ${\mu}$M) for 24 h incubation. These results suggest that THP at 5-10${\mu}$M decreases the basal dopamine content and reduces the increased dopamine content induced by L-DOPA in part by the inhibition of TH activity, and that THP at 15${\mu}$M also decreases dopamine content by oxidative stress in PC12 cells.

• 대한화장품학회지
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• 제36권1호
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• pp.79-87
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• 2010

자외선은 생체 내 활성산소 생성 증가와 내인성 항산화 효소 및 항산화제 감소를 통하여 광노화 과정을 촉진하는 대표적인 외부 환경인자로 알려져 있다. 그러므로 본 연구에서는 12종의 다양한 해양소재 추출물(김, 다시마, 모려, 모자반, 미역, 석결명, 청각, 해룡, 해마, 해삼, 파래, 톳)을 이용하여 기본적인 자유라디칼 소거능 실험을 바탕으로, 자외선 B 조사로 인한 세포독성 및 산화적 사멸을 억제하고 세포 내 항산화 기능을 강화시킬 수 있는 천연 항산화 피부보호소재를 발굴하고자 하였다. 해양소재 추출물의 라디칼 소거능을 1,1-diphenyl-2-picrylhydrazyl (DppH) assay로 비교 측정한 결과 미역, 모자반, 다시마, 해마, 석결명 모려, 해룡의 순으로 항산화력을 나타내었다. 이후 in vitro 세포 실험에서 자외선 B를 섬유아세포주인 HS68 세포에 노출시킨 경우 세포 독성이 유발되어 사멸이 진행되었으며 이는 모자반, 석결명, 청각, 해마 열수 추출물을 처리한 경우 현저히 억제되었다. 특히, 자외선 B로 인한 세포손상은 세포 내 활성산소종의 축적으로 인한 것임을 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA) 형광염색법으로 확인할 수 있었으며, 활성산소종의 생성은 모자반, 석결명, 청각, 해마 열수 추출물을 처리한 경우 유의적으로 감소되었다. 한편, 이러한 항산화 작용은 대표적인 내인성 항산화 효소인 catalase, superoxide dismutase 및 heme oxygenase-1의 발현 증가로 매개되었다. 이상의 결과는 모자만, 석결명, 정각, 해마 풍의 해양소재가 항산화 작용을 통하여 산화적 스트레스가 매개하는 피부손상과 노화과정을 예방 및 보호하는 새로운 화장품 천연소재로의 활용 가능성을 제시한다.

• The Korean Journal of Physiology and Pharmacology
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• 제21권2호
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• pp.233-239
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• 2017

Intracellular calcium ($Ca^{2+}$) oscillation is an initial event in digestive enzyme secretion of pancreatic acinar cells. Reactive oxygen species are known to be associated with a variety of oxidative stress-induced cellular disorders including pancreatitis. In this study, we investigated the effect of hydrogen peroxide ($H_2O_2$) on intracellular $Ca^{2+}$ accumulation in mouse pancreatic acinar cells. Perfusion of $H_2O_2$ at $300{\mu}M$ resulted in additional elevation of intracellular $Ca^{2+}$ levels and termination of oscillatory $Ca^{2+}$ signals induced by carbamylcholine (CCh) in the presence of normal extracellular $Ca^{2+}$. Antioxidants, catalase or DTT, completely prevented $H_2O_2$-induced additional $Ca^{2+}$ increase and termination of $Ca^{2+}$ oscillation. In $Ca^{2+}$-free medium, $H_2O_2$ still enhanced CCh-induced intracellular $Ca^{2+}$ levels and thapsigargin (TG) mimicked $H_2O_2$-induced cytosolic $Ca^{2+}$ increase. Furthermore, $H_2O_2$-induced elevation of intracellular $Ca^{2+}$ levels was abolished under sarco/endoplasmic reticulum $Ca^{2+}$ ATPase-inactivated condition by TG pretreatment with CCh. $H_2O_2$ at $300{\mu}M$ failed to affect store-operated $Ca^{2+}$ entry or $Ca^{2+}$ extrusion through plasma membrane. Additionally, ruthenium red, a mitochondrial $Ca^{2+}$ uniporter blocker, failed to attenuate $H_2O_2$-induced intracellular $Ca^{2+}$ elevation. These results provide evidence that excessive generation of $H_2O_2$ in pathological conditions could accumulate intracellular $Ca^{2+}$ by attenuating refilling of internal $Ca^{2+}$ stores rather than by inhibiting $Ca^{2+}$ extrusion to extracellular fluid or enhancing $Ca^{2+}$ mobilization from extracellular medium in mouse pancreatic acinar cells.

• Journal of Microbiology and Biotechnology
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• 제29권4호
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• pp.518-526
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• 2019

Pueraria montana var. lobata is a bioactive substance with various beneficial health effects and has long been extensively used as a traditional medication for the treatment of fever, acute dysentery, diabetes, and cardiovascular diseases in Northeast Asian countries. The purpose of this study was to evaluate the cytoprotective activity of Pueraria montana var. lobata ethanol extract (PLE) for ultraviolet B (UVB)-induced oxidative stress in human dermal fibroblasts (HDF). It was hypothesized that PLE treatment ($25-100{\mu}g/ml$) would reduce intracellular reactive oxygen species (ROS) levels as well as increase collagen production in UVB-irradiated HDF. The results confirmed this theory, with collagen production increasing in the PLE treatment group in a dose-dependent manner. In addition, regulators of cellular ROS accumulation, including HO-1 and NOQ-1, were activated by Nrf2, which was mediated by PLE. Hence, intracellular levels of ROS were also reduced in the PLE treatment group in a dose-dependent manner. In conclusion, PLE increases collagen production and maintains hyaluronic acid (HA) levels in human dermal fibroblasts exposed to UVB-irradiation, thereby inhibiting photoaging.