• Animal Bioscience
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• 제34권6호
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• pp.1006-1013
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• 2021

Objective: Cells have increased susceptibility to activation of apoptosis when suffering heat stress (HS). An effective supplementation strategy to mimic heat-induced apoptosis of bovine mammary epithelial cells (MECs) is necessary to maintain optimal milk production. This study aimed to investigate possible protective effects of the anti-apoptotic activity of 5-aminolevulinic acid (5-ALA) against HS-induced damage of bovine MECs. Methods: Bovine MECs were pretreated with or without 5-ALA at concentrations of 10, 100, and 500 µM for 24 h followed by HS (42.5℃ for 24 h and 48 h). Cell viability was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Real-time quantitative polymerase chain reaction and Western blotting were used to explore the regulation of genes associated with apoptosis, oxidative stress, and endoplasmic reticulum (ER) stress genes. Results: We found that 5-ALA induces cytoprotection via inhibition of apoptosis markers after HS-induced damage. Pretreatment of bovine MECs with 5-ALA resulted in dramatic upregulation of mRNA for nuclear factor erythroid-derived 2-like factor 2, heme oxygenase-1, and NAD(P)H quinone oxidoreductase 1, all of which are antioxidant stress genes. Moreover, 5-ALA pretreatment significantly suppressed HS-induced ER stress-associated markers, glucose-regulated protein 78, and C/EBP homologous protein expression levels. Conclusion: 5-ALA can ameliorate the ER stress in heat stressed bovine MEC via enhancing the expression of antioxidant gene.

• 대한화장품학회지
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• 제39권1호
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• pp.19-24
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• 2013

본 연구에서는 감태 효소 추출물과 그것의 폴리페놀 추출물의 화장품 원료로서의 효능을 알아보기 위하여 항산화, 항당화, 미백, 항염 효과와 관련된 실험을 실시하였다. 감태 효소 추출물과 폴리페놀 추출물은 강력한 라디컬 소거능을 가지고 있으며 BSA/Glucose 시스템에서 최종당화생성물의 형성을 저해하는 항당화 활성과 타이로시네이즈 저해를 통한 우수한 미백력을 가지고 있음을 확인하였다. 또한 두 추출물 모두 세포 내에서 $PGE_2$와 NO 생성 저해를 통한 항염 효과를 나타내었다. 이러한 결과를 종합해 볼 때, 감태 효소 추출물과 그 폴리페놀 추출물은 화장품 원료로서의 응용 가능성이 있을 것으로 사료된다.

• 대한본초학회지
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• 제30권5호
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• pp.23-28
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• 2015

Objectives : The aim of this study is to investigate cell viability, anti-inflammatory, antioxidant, immunoenhancing activity using various extracts of Yeonsan Ogye.Methods : In order to evaluate cytotoxicity, MTT assay was performed. We investigated production levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-αand interleukin (IL)-6, and nitric oxide(NO) in LPS-induced RAW 264.7 cells. NO production in RAW 264.7 cells was measured by using Griess reagent. Cytokines including IL-6 and TNF-αwere measured by Luminex and ROS was measured by Flow cytometry.Results : No cytotoxicity of various extracts of Yeonsan Ogye was observed in RAW 264.7 cells. Productions of ROS in RAW 264.7 cells were increased from extraction of bones and decreased from extraction of skin. Also, productions of NO in RAW 264.7 cells were increased to bone extract and decreased at skin extract. In addition, productions of pro-inflammatory cytokines (IL-6 and TNF-α) in LPS-induced RAW 264.7 cells were decreased at skin, meat extracts, respectively. Finally, the levels of immune-related cytokines (IL-6 and TNF-α) were increased compared to those of the normal group.Conclusions : It is concluded that Yeonsan Ogye extracts seem to have significant biological activities likes anti-inflammatory, antioxidant, immuno-enhancing etc. These results may be developed as a raw material for new health food and new therapeutics to ease the symptoms related with inflammatory and oxidative stress. In terms of oriental traditional medicine, we expect that it contribute to building of EBM (Evidence-Based Medicine) from the this result.

• ALGAE
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• 제30권4호
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• pp.313-323
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• 2015

Lipid peroxidation means the oxidative degradation of lipids. The process from the cell membrane lipids in an organism is generated by free radicals, and result in cell damage. Phlorotannins, well-known marine brown algal polyphenols, have been utilized in functional food supplements as well as in medicine supplements to serve a variety of purposes. In this study, we assessed the potential anti-lipid peroxidation activity of phlorofucofuroeckol-A (PFF-A), one of the phlorotannins, isolated from Ecklonia cava by centrifugal partition chromatography in 2,2-azobis (2-amidinopropane) dihydrochloride (AAPH)-stimulated Vero cells and zebrafish system. PFF-A showed the strongest scavenging activity against alkyl radicals of all other reactive oxygen species (ROS) and exhibited a strong protective effect against ROS and a significantly strong inhibited of malondialdehyde in AAPH-stimulated Vero cells. The apoptotic bodies and pro-apoptotic proteins Bax and caspase-3, which were induced by AAPH, were strongly inhibited by PFF-A in a dose-dependent manner and expression of Bcl-xL, an anti-apoptotic protein, was induced. In the AAPH-stimulated zebrafish model, additionally PFF-A significantly inhibited ROS and cell death, as well as exhibited a strong protective effect against lipid peroxidation. Therefore, these results suggest that PFF-A has excellent protective effects against ROS and lipid peroxidation induced by AAPH in both an in vitro Vero cell model and an in vivo zebrafish model.

• 한방재활의학과학회지
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• 제25권2호
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• pp.15-35
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• 2015

Objectives This study was performed to investigate the effects of Bujasasim-tang ethanol extract (BST) on oxidative stress, inflammation and osteoarthritic rat model. Methods To ensure safety of BST, heavy metal levels were measured and cytotoxicity test was done. In vitro, To evaluate antioxidative effects of BST, total phenolic contents, 1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) scavenging activity, reactive oxygen species (ROS) levels were measured. Also, to evaluate anti-inflammatory effects of BST treated group, total nitric oxide (NO) and pro-inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$) levels were measured in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In vivo, We injected MIA $50{\mu}l$ (60 mg/ml) into knee joints of rats to induce osteoarthritis. Rats were divided into total 3 groups (normal, control, BST treated group, each n=7). Normal group was not treated at all without inducing osteoarthritis and taken normal diet. Control group was induced osteoarthritis by MIA and taken with 2 ml of distilled water once a day for 4 weeks. BST treated group was induced osteoarthritis by MIA and taken BST 2 ml (200 mg/kg/mouse) once a day for 4 weeks. We evaluated dynamic weight bearing with the Incapacitance Test Meter. At the end of experiment, the rats were sacrificed to observe the functions of liver and kidney, changes of WBC, neutrophil, lymphocyte, monocyte levels in blood, to evaluate the levels of pro-inflammatory cytokines, tissue inhibitor of metallopreteinases-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), prostaglandin $E_2$ ($PGE_2$), leukotriene $B_4$ ($LTB_4$) within serum. We observed change of articular structures by Hematoxylin & Eosin (H&E), safranin-O staining method and measured amount of cartilage by micro CT-arthrography. Statistical analysis was done by unpaired student's t-test with significance level at p<0.05 in SPSS 11.0 for windows. Results 1. Safety of the BST was identified. 2. AST, ALT, BUN, creatinine levels of BST treated group were within normal limit. In vitro, 1. DPPH and ABTS free radical scavenging activities of BST showed dose-dependent increase. 2. ROS production were significantly decreased. 3. Total nitric oxide (NO) and IL-$1{\beta}$ production were decreased. 4. IL-6 and TNF-${\alpha}$ production were significantly decreased. In vivo, 1. Weight bearing ability was significantly increased. 2. WBC, neutrophil, lymphocyte, monocyte levels in blood were decreased. 3. IL-$1{\beta}$ and TNF-${\alpha}$ levels in serum were significantly decreased. and the IL-6 level was decreased. 4. TIMP-1, MMP-9, $LTB_4$, $PGE_2$ levels in serum were significantly decreased. 5. Cartilage volume of BST treated group was significantly increased. Also changes of cartilage, synovial membrane, fibrous tissue were suppressed. Conclusions The results obtained in this study Bujasasim-tang have effects of antioxidative, anti-inflammatory, relieve pain and protection of cartilage. Therefore we expect that Bujasasim-tang is effective treatment for osteoarthritis.

• Biomolecules & Therapeutics
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• 제28권2호
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• pp.152-162
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• 2020

Cerebral ischemia exhibits a multiplicity of pathophysiological mechanisms. During ischemic stroke, the reactive oxygen species (ROS) concentration rises to a peak during reperfusion, possibly underlying neuronal death. Recombinant human erythropoietin (EPO) supplementation is one method of treating neurodegenerative disease by reducing the generation of ROS. We investigated the therapeutic effect of PEGylated EPO (P-EPO) on ischemic stroke. Mice were administered P-EPO (5,000 U/kg) via intravenous injection, and middle cerebral artery occlusion (MCAO) followed by reperfusion was performed to induce in vivo ischemic stroke. P-EPO ameliorated MCAO-induced neurological deficit and reduced behavioral disorder and the infarct area. Moreover, lipid peroxidation, expression of inflammatory proteins (cyclooxygenase-2 and inducible nitric oxide synthase), and cytokine levels in blood were reduced by the P-EPO treatment. In addition, higher activation of nuclear factor kappa B (NF-κB) was found in the brain after MCAO, but NF-κB activation was reduced in the P-EPO-injected group. Treatment with the NF-κB inhibitor PS-1145 (5 mg/kg) abolished the P-EPO-induced reduction of infarct volume, neuronal death, neuroinflammation, and oxidative stress. Moreover, P-EPO was more effective than EPO (5,000 U/kg) and similar to a tissue plasminogen activator (10 mg/kg). An in vitro study revealed that P-EPO (25, 50, and 100 U/mL) treatment protected against rotenone (100 nM)-induced neuronal loss, neuroinflammation, oxidative stress, and NF-κB activity. These results indicate that the administration of P-EPO exerted neuroprotective effects on cerebral ischemia damage through anti-oxidant and anti-inflammatory properties by inhibiting NF-κB activation.

• 대한한방부인과학회지
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• 제20권2호
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• pp.139-154
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• 2007

Purpose: This study examined the Cell differentiation and the anti-oxidative effect of Dioscoreae Rhizoma on HeLa cells. We are interested in whether Dioscoreae Rhizoma is capable of causing apoptosis processes on HeLa cell, and whether cotreatment of NCS with Dioscoreae Rhizoma reduces cell viability. Methods: We used aqueous extract to treat HeLa cell with different concentrations treated with a water or a MeOH extract of Dioscoreae Rhizoma (0, x10, x20, x40, x80). The MTT (3, (4, 5-dimethyl-thiazol) 2, 5-diphenyl-tetraxolium bromide) reduction assay was employed to quantify the differences in cell activity and viability. Cells were stained with DAPI and visualized by fluorescent Microscope. The caspase-3, Bcl-2, PARP, p53 expression level was monitored using western-blotting techniques. The patterns of the changes in expression were scanned and analyzed. Results: The survival rate of cells treated with Dioscoreae Rhizoma extracts increased by 20% at specific concentration. The other side Dioscoreae Rhizoma extracts induced apoptotic features including chromatin condensation and fragmentation. And Dioscoreae Rhizoma extracts increased the expression of caspase-3, p53 and the cleavage of PARP protein. However, co-treatment with Dioscoreae Rhizoma with NCS attenuated the activations of p53 and PARP protein that were mediated by NCS treatment alone. This is indicated Dioscoreae batatas extracts attenuated cytotoxicity induced by oxidative agents including NCS. Conclusion: Our results suggest Dioscoreae Rhizoma extracts induce cell differentiation or apoptosis connected with concentration. Further elucidation of concentration of Dioscoreae Rhizoma awaits many other biochemical investigative studies.

• 한국잠사곤충학회지
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• 제47권2호
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• pp.62-67
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• 2005

장려뽕 16 품종에 대한 뽕잎과 오디의 항산화능을 시기별로 채취하여 비교분석 하였다. 항산화능 측정장치(munilum L-100, ABCD GmbH) 및 ARAW-KIT(antiradical ability of water-soluble substance)를 사용하였으며, ascorbic acid를 표준물질로 사용하여 calibration curve를 작성하였다. 시료의 항산화능과 분석에 소요되는 전처리 시간을 고려하여 80% MeOH, 30 sec. vortex mixing 방법을 기준 전처리 방법으로 하였다. 16품종 전체의 뽕잎에 대한 항산화능을 분석한 결과 5개엽기의 어린 뽕잎의 항산화능은 3303.4 nmol로 높았으며, 오디가 착색되기 직전 3708.0 nmol으로 최대 값을 나타낸 후 시기가 경과함에 따라 점차 감소 및 하벌 후 점차 증가 하다가 뽕잎이 경화되기 직전 춘기 어린 잎 정도의 항산화능을 유지하는 것으로 나타났다. 항산화능이 높은 뽕잎을 이용하기 위해서는 5월 하순에 채취하는 것이 수율도 높아 경제성이 있을 것으로 판단되며, 생과 또는 가공식품의 원료로 사용하기 위해 오디를 수확한 후 남은 뽕잎의 이용도 충분히 가능하므로 이를 활용하기 위해서는 하벌 전에 뽕잎의 수확 작업이 이루어지도록 해야 할 것이다. 뽕잎 수확 후 저장하는 방법보다 시기에 따른 뽕잎 자체의 성숙 정도가 뽕잎의 항산화능을 결정하는 것으로 판단된다.

• 한국식품영양과학회지
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• 제44권12호
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• pp.1832-1838
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• 2015

${\omega}-3$ 지방산이 풍부한 들깨를 이용하여 고령자용 건강간식을 만들기 위해 다식을 제조하고 들깨의 산패 억제를 위해 당근가루를 0, 5, 10 15, 20% 첨가하여 들깨가루 다식을 제조한 결과, 당근가루 첨가량에 따른 들깨 다식의 수분 함량의 변화는 없었으며, 당근 내 카로티노이드에 의해 당근가루 첨가량이 많아질수록 다식의 붉은색과 황색도는 증가하였다. 당근가루 첨가량이 많을수록 다식의 경도와 응집성, 씹힘성은 증가하였다. 다식의 과산화물가와 산가를 측정한 결과 당근가루 첨가량이 많을수록 항산화 효과는 증가하였고, 라디칼 소거능 역시 효과적으로 증가하였다. 지방을 함유한 식품에는 산패 억제를 위해 항산화제를 사용하고 있으며, 특히 합성 항산화제를 대체할 천연 항산화제에 대한 요구가 계속적으로 증가하고 있는데 열처리(데치기)에 의해 ${\beta}-carotene$을 활성화시킨 당근가루는 들깨다식의 항산화 활성을 증진시켜 질적 향상을 유도할 수 있는 좋은 천연 항 산화제로서의 기능을 갖고 있는 것으로 조사되었다. 관능검사 결과 당근가루 첨가 여부에 대한 선호는 뚜렷했으나, 첨가 수준에 따른 선호도의 차이는 없었다. 본 연구 결과 20% 수준의 당근가루 첨가는 항산화 특성을 효과적으로 향상시키면서 전체적인 기호도에 영향을 미치지 않아 들깨다식 제조 시 적정 첨가 수준으로 생각된다.

• Toxicological Research
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• 제31권1호
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• pp.17-23
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• 2015

Excessive manganese (Mn) in the brain promotes a variety of abnormal behaviors, including memory deficits, decreased motor skills and psychotic behavior resembling Parkinson's disease. Hereditary hemochromatosis (HH) is a prevalent genetic iron overload disorder worldwide. Dysfunction in HFE gene is the major cause of HH. Our previous study has demonstrated that olfactory Mn uptake is altered by HFE deficiency, suggesting that loss of HFE function could alter manganese-associated neurotoxicity. To test this hypothesis, Hfe-knockout ($Hfe^{-/-}$) and wild-type ($Hfe^{+/+}$) mice were intranasally-instilled with manganese chloride ($MnCl_2$ 5 mg/kg) or water daily for 3 weeks and examined for memory function. Olfactory Mn diminished both short-term recognition and spatial memory in $Hfe^{+/+}$ mice, as examined by novel object recognition task and Barnes maze test, respectively. Interestingly, $Hfe^{-/-}$ mice did not show impaired recognition memory caused by Mn exposure, suggesting a potential protective effect of Hfe deficiency against Mn-induced memory deficits. Since many of the neurotoxic effects of manganese are thought to result from increased oxidative stress, we quantified activities of anti-oxidant enzymes in the prefrontal cortex (PFC). Mn instillation decreased superoxide dismutase 1 (SOD1) activity in $Hfe^{+/+}$ mice, but not in $Hfe^{-/-}$ mice. In addition, Hfe deficiency up-regulated SOD1 and glutathione peroxidase activities. These results suggest a beneficial role of Hfe deficiency in attenuating Mn-induced oxidative stress in the PFC. Furthermore, Mn exposure reduced nicotinic acetylcholine receptor levels in the PFC, indicating that blunted acetylcholine signaling could contribute to impaired memory associated with intranasal manganese. Together, our model suggests that disrupted cholinergic system in the brain is involved in airborne Mn-induced memory deficits and loss of HFE function could in part prevent memory loss via a potential up-regulation of anti-oxidant enzymes in the PFC.