• BMB Reports
• /
• 제40권6호
• /
• pp.928-935
• /
• 2007

Three Angelica sinensis polysaccharide fractions (APFs), named APF1, APF2 and APF3, were isolated and purified from Radix A. sinensis and their antioxidant activities were evaluated in isolated mouse peritoneal macrophages by pretreatment with APFs before exposure to 0.2 mM tertbutylhydroperoxide (t-BHP). The results showed that pretreatment of the macrophages with APFs as low as $10{\mu}g$/ml could significantly enhance t-BHP-decreased cell survival, intracellular glutathione (GSH) content and superoxide dismutase (SOD) activity, and also inhibited t-BHP-increased lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation (p < 0.05), and APF3 was the most active fraction, followed by APF2 and APF1 in decreasing order. Furthermore, we found for the first time that the bound-protein in APF3 was associated closely with the protective effects and the polysaccharide inhibited the excess NO release from t-BHP-activated macrophages to protect host cells.

• Journal of Ginseng Research
• /
• 제36권3호
• /
• pp.248-255
• /
• 2012

Potential antioxidant effect of processed ginseng (sun ginseng, SG) on oxidative stress generated by tert-butyl hydroperoxide (t-BHP) was investigated in HepG2 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase (LDH) leakage test demonstrated that SG dose-dependently prevents a loss of cell viability against t-BHP-induced oxidative stress. Also, SG treatment dose-dependently relieved the increment of activities of hepatic enzymes, such as aspartate aminotrasferase and alanine aminotransferase, and lipid peroxidation mediated by t-BHP treatment in HepG2 cells. SG increased the gene expression of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. However, high dose of SG treatment caused decrease in mRNA level of glutathione peroxidase as compared to low dosage of SG-treated cells. The gene expression of glutathione reductase was found to be slightly increased by SG treatment. In addition, SG extract attributed its hepaprotective effect by inducing the mRNA level of bcl-2 and bcl-xL but reducing that of bax. But, the gene expression of bad showed no significant change in SG-treated HepG2 cells. These findings suggest that SG has hepatoprotective effect by showing reduction of LDH release, activities of hepatic enzymes and lipid peroxidation and regulating the gene expression of antioxidant enzymes and apoptosis-related molecules against oxdative stress caused by t-BHP in HepG2 cells.

• Nutrition Research and Practice
• /
• 제11권2호
• /
• pp.97-104
• /
• 2017

BACKGROUND/OBJECTIVE: Although Angelica keiskei (AK) has widely been utilized for the purpose of general health improvement among Asian, its functionality and mechanism of action. The aim of this study was to determine the protective effect of ethanol extract of AK (AK-Ex) on acute hepatotoxicity induced by acetaminophen (AAP) in HepG2 human hepatocellular liver carcinoma cells and HepaRG human hepatic progenitor cells. MATERIALS/METHODS: AK-Ex was prepared HepG2 and HepaRG cells were cultured with various concentrations and 30 mM AAP. The protective effects of AK-Ex against AAP-induced hepatotoxicity in HepG2 and HepaRG cells were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, lactate dehydrogenase (LDH) assay, flow cytometry, and Western blotting. RESULTS: AK-Ex, when administered prior to AAP, increased cell growth and decreased leakage of LDH in a dose-dependent manner in HepG2 and HepaRG cells against AAP-induced hepatotoxicity. AK-Ex increased the level of Bcl-2 and decreased the levels of Bax, Bok and Bik decreased the permeability of the mitochondrial membrane in HepG2 cells intoxicated with AAP. AK-Ex decreased the cleavage of poly (ADP-ribose) polymerase (PARP) and the activation of caspase-9, -7, and -3. CONCLUSIONS: These results demonstrate that AK-Ex downregulates apoptosis via intrinsic and extrinsic pathways against AAP-induced hepatotoxicity. We suggest that AK could be a useful preventive agent against AAP-induced apoptosis in hepatocytes.

• Journal of Microbiology and Biotechnology
• /
• 제24권1호
• /
• pp.87-96
• /
• 2014

In the present study, a yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp. VITGBN2 based on molecular techniques and was found to be a potent producer of acidic diacetate sophorolipid in mineral salt media containing vegetable oil as additional carbon source. The chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. This sophorolipid was used as a stabilizer for synthesis of zinc oxide nanoparticles (ZON). The formation of biofunctionalized ZON was characterized using UV-visible spectroscopy, XRD, scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy. The antimicrobial activities of naked ZON and sophorolipid functionalized ZON were tested based on the diameter of inhibition zone in agar well diffusion assay, microbial growth rate determination, protein leakage analysis, and lactate dehydrogenase assay. Bacterial pathogen Salmonella enterica and fungal pathogen Candida albicans showed more sensitivity to sophorolipid biofunctionalized ZON compared with naked ZON. Among the two pathogens, S. enterica showed higher sensitivity towards sophorolipid biofunctionalized ZON. SEM analysis showed that cell damage occurred through cell elongation in the case of S. enterica, whereas cell rupture was found to occur predominantly in the case of C. albicans. This is the first report on the dual role of yeast-mediated sophorolipid used as a biostabilizer for ZON synthesis as well as a novel functionalizing agent showing antimicrobial property.

• The Korean Journal of Physiology and Pharmacology
• /
• 제19권4호
• /
• pp.357-363
• /
• 2015

The purpose of this study was to compare the inhibitory effect of bevacizumab on human Tenon's fibroblasts (HTFs) cultured from primary and recurrent pterygium. Cultured HTFs were exposed to 2.0, 5.0, 7.5, and 15.0 mg/mL concentration of bevacizumab for 24 hours. The 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase leakage assays were then performed to assess fibroblast metabolism and viability. The matrix metalloproteinase (MMP), procollagen type I C terminal propeptide (PIP), and laminin immunoassays were performed to examine extracellular matrix production. Changes in cellular morphology were examined by phase-contrast and transmission electron microscopy. Both metabolic activity and viability of primary and recurrent pterygium HTFs were inhibited by bevacizumab in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. Both types of HTFs had significant decreases in MMP-1, PIP, and laminin levels. Distinctly, the inhibitory effect of bevacizumab on MMP-1 level related with collagenase in primary pterygium HTFs was significantly higher than that of recurrent pterygium. Significant changes in cellular density and morphology both occurred at bevacizumab concentrations greater than 7.5 mg/mL. Only primary pterygium HTFs had a reduction in cellular density at a bevacizumab concentration of 5.0 mg/mL. Bevacizumab inhibits primary and recurrent pterygium HTFs in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. As the primary HTFs produces larger amounts of MMP-1 compared to recurrent HTFs, significant reduction in MMP-1 level in primary pterygium HTFs after exposure to bevacizumab is likely to be related to the faster cellular density changes in primary pterygium HTFs.

• 한국식품영양과학회지
• /
• 제35권9호
• /
• pp.1121-1126
• /
• 2006

In the present study, the potential hepatoprotective effects of poly herbal formulation, Hepa-1000, against oxidative damages induced by t-BHP were evaluated in HepG2 cells in order to relate in vitro antioxidant activity with cytoprotective effects. The t-BHP induced considerable cell damage in HepG2 cells was shown by significant glutamic oxaloacetic transaminase (GOT) and lactate dehydrogenase (LDH) leakage, and increased lipid peroxidation. Hepa-1000-treated cells showed an increased resistance to oxidative challenge, as revealed by higher survival capacity than the one of control cells against t-BHP induced oxidative stress and hepatotoxicity. In addition, the Hepa-1000 had hepatoprotective effects lowering the activity of GOT and LDH, simultaneously. That is, it could inhibit the cell membrane damages resulting in the increased activities of GOT and LDH in the cell culture media. Furthermore, the Hepa-1000 could reduce t-BHP enhanced lipid peroxidation, which was evaluated by measuring the production of malonedialdehyde. Based on the data described above, it could be suggested that the Hepa-1000 has significant hepatoprotective effects and plays a protective role against lipid peroxidation by free radicals.

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

• 한국자원식물학회지
• /
• 제25권2호
• /
• pp.184-192
• /
• 2012

본 연구는 췌장베타세포인 HIT-T15 세포를 이용하여 매실주정추출물(PME)의 alloxan에 의한 산화스트레스로부터의 세포보호, 인슐린 분비능 및 항산화 효소 활성을 평가하였다. PME는 alloxan에 의해 유발된 산화스트레스로부터 세포를 보호하여 세포생존율을 증가시켰다. PME는 세포막 손상지표인 LDH 방출을 억제하였고 $NAD^+$/NADH ratio를 유의적으로 증가시켜 세포사멸이 억제되어짐을 확인하였다. 또한 alloxan 단독처리군에 비해 250 ${\mu}g$/ml PME 처리군에서 인슐린 분비량이 유의성 있게 증가하였다. Alloxan과 PME를 동시에 처리하여 HIT-T15세포의 항산화효소 활성을 측정했을 때 산화스트레스에 의해 감소되었던 항산화효소 활성이 PME에 의해 보호되는 효과를 확인하였다. 이상의 연구결과로부터 PME는 세포괴사 및 DNA fragmentation을 억제하고 세포내 항산화효소 활성을 증가시켜 alloxan에 의해 유발된 산화스트레스로부터 췌장베타세포를 보호하고 이에 따른 인슐린 분비능 조절 효과가 있는 것으로 생각된다.

• 한국미생물·생명공학회지
• /
• 제39권1호
• /
• pp.77-85
• /
• 2011

본 연구는 은나노 입자의 항균활성을 알아보기 위하여, 그람 양성세균인 황색포도상구균과 그람 음성세균인 대장균에 대한 은나노 입자(Ag-NPs)를 처리 후, 세균세포 생장곡선측정, 활성산소생성능 측정, 세포질 단백질 누출량 측정, 젖산탈수소효소 활성측정 및 고분해능 임계방사 주사전자현미경 관찰이 수행되었다. 세균세포의 생장곡선 측정은 다양한 농도, 배양시간, 배양온도 및 pH에서 수행되었다. 결과적으로 황색 포도상구균과 대장균은 배양온도와 pH에 영향을 받지않고 은나노 입자에 의해 효과적으로 생장억제가 이루어지는 것을 관찰할 수 있었다. 또한 활성산소의 생성에 의하여 세포막의 파괴로 세포질내 물질의 유출을 세포질 유래 단백질 측정으로 확인할 수 있었으며, 젖산탈수소효소 활성측정을 통하여 은나노 입자에 대한 세포호흡억제활성 또한 확인할 수 있었다. 임계방사 주사전자현미경 관찰결과 은나노 입자에 의한 세균 세포표면의 형태학적 변화 또한 관찰되었다. 이러한 결과를 통하여 은나노 입자를 효과적인 항균활성소재로 활용 가능함이 입증되었다.

• 한국식품과학회지
• /
• 제52권5호
• /
• pp.546-554
• /
• 2020

본 연구에서는 국내에서 다빈도로 사용되고 있는 천연색소, 천연추출물 및 기타용도 천연첨가물 10종에 대한 독성 영향을 세포 수준에서 확인하였다. 세포 처리 농도 및 시간은 품목제조보고를 바탕으로 확인한 천연첨가물 다빈도 첨가 품목 및 그 최대사용량과 국민영양통계 일일섭취량을 기반으로 소장액 부피 및 소장 내 체류시간을 종합적으로 고려하여 설정하였다. 세포 독성 시험 결과, 10종 천연첨가물에 의한 세포성장 저해 및 사멸 유발 영향이 확인되지 않았으며 세포 내 활성산소종이 유발되지 않는 한편, 활성산소종 소거능을 보유하고 있는 것으로 나타났다. 또한, 젖산탈수소효소 분석을 통해 세포막 손상을 유발하지 않는 것으로 확인되었다. 체내 장관계와 유사한 환경을 모사한 2차원 및 3차원 장관계 모사모델을 통해 세포성장 및 사멸 효과 확인 결과, 본 연구에서 사용한 10종의 천연첨가물에 의해 세포성장 저해 및 사멸이 유발되지 않았다. 종합적으로, 본 연구에서 확인한 다빈도로 사용되는 천연첨가물 10종의 세포독성이 현재 사용수준에서는 낮은 것으로 판단되어 안전한 것으로 보인다. 이와 같은 연구결과는 안전성에 대한 정보가 상대적으로 미흡했던 천연첨가물에 대한 구체적인 독성자료로 활용될 수 있으며, 향후 보다 심도 깊은 in vivo 독성 연구를 위한 기초 토대를 마련할 것으로 기대된다.