• Title/Summary/Keyword: Glutathione Production

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Hepato-Protective Activities of Jasminum officinale L. var. grandiflorum Aqueous Extract via Activation of AMPK in HepG2 Cells (AMPK 활성화를 통한 소형화(素馨花) (Jasminum officinale L. var. grandiflorum) 열수 추출물의 HepG2 간세포 보호 활성)

  • Sang Mi Park;Dae Hwa Jung;Byung Gu Min;Kyung Hwan Jegal;Sung Hui Byun;Jae Kwang Kim;Sang Chan Kim
    • Herbal Formula Science
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    • v.31 no.4
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    • pp.231-243
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    • 2023
  • Objectives : Jasminum officinale L. var. grandiflorum is used as a traditional or folk remedy in China to treat arthritis, hepatitis, duodenitis, conjunctivitis, gastritis, and diarrhea. In this study, we aimed to study the hepatocyte protective activity and molecular mechanism of Jasminum officinale L. var. grandiflorum aqueous extract (JGW) using HepG2 hepatocyte cell lines. Methods : HepG2 cells were pretreated with diverse concentrations of JGW, and then the cells were exposed to tert-butyl hydroperoxide (tBHP) for inducing oxidative stress. Hydrogen peroxide (H2O2) production, glutathione (GSH) concentration, mitochondrial membrane potential (MMP) and cell viability were measured to investigate hepato-protective effects of JGW. Phosphorylation of AMP-activated protein kinases (AMPK), acetyl coenzyme A carboxylase (ACC) and effects of compound C on cell viability were examined to observe the role of AMPK on JGW-mediated cytoprotection. Results : Pretreatment with JGW (10-300 ㎍/mL) significantly suppressed cytotoxicity induced by tBHP in a concentration dependent manner and reduced the expression of cleaved PARP and cleaved caspase-3 proteins related to apoptosis in HepG2 cells. In addition, pretreatment with JGW significantly prevented the increase in H2O2 production, GSH depletion, and lower MMP induced by tBHP. Treatment with JGW (30 minutes of incubation and concentrations of 100 and 300 ㎍/mL) increased the phosphorylation of AMPK and ACC and treatment with compound C, a chemical inhibitor of AMPK, inhibited the cytoprotective effect of JGW. Conclusions : Our results demonstrated that JGW may protect hepatocytes from oxidative stress via activation of AMPK.

Morroniside Protects C2C12 Myoblasts from Oxidative Damage Caused by ROS-Mediated Mitochondrial Damage and Induction of Endoplasmic Reticulum Stress

  • Hyun Hwangbo;Cheol Park;EunJin Bang;Hyuk Soon Kim;Sung-Jin Bae;Eunjeong Kim;Youngmi Jung;Sun-Hee Leem;Young Rok Seo;Su Hyun Hong;Gi-Young Kim;Jin Won Hyun;Yung Hyun Choi
    • Biomolecules & Therapeutics
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    • v.32 no.3
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    • pp.349-360
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    • 2024
  • Oxidative stress contributes to the onset of chronic diseases in various organs, including muscles. Morroniside, a type of iridoid glycoside contained in Cornus officinalis, is reported to have advantages as a natural compound that prevents various diseases. However, the question of whether this phytochemical exerts any inhibitory effect against oxidative stress in muscle cells has not been well reported. Therefore, the current study aimed to evaluate whether morroniside can protect against oxidative damage induced by hydrogen peroxide (H2O2) in murine C2C12 myoblasts. Our results demonstrate that morroniside pretreatment was able to inhibit cytotoxicity while suppressing H2O2-induced DNA damage and apoptosis. Morroniside also significantly improved the antioxidant capacity in H2O2-challenged C2C12 cells by blocking the production of cellular reactive oxygen species and mitochondrial superoxide and increasing glutathione production. In addition, H2O2-induced mitochondrial damage and endoplasmic reticulum (ER) stress were effectively attenuated by morroniside pretreatment, inhibiting cytoplasmic leakage of cytochrome c and expression of ER stress-related proteins. Furthermore, morroniside neutralized H2O2-mediated calcium (Ca2+) overload in mitochondria and mitigated the expression of calpains, cytosolic Ca2+-dependent proteases. Collectively, these findings demonstrate that morroniside protected against mitochondrial impairment and Ca2+-mediated ER stress by minimizing oxidative stress, thereby inhibiting H2O2-induced cytotoxicity in C2C12 myoblasts.

Dietary Effects of Fermented Soybean Curd Residue (Biji) on Body Weight, Serum Lipid Profiles, and Antioxidation-Related Enzymes Activity of Mice Fed a High Fat Diet (고지방식이 마우스의 체중과 혈청지질 및 항산화계 효소활성에 미치는 발효 비지의 식이효과)

  • Lee, Sang-Il;Lee, Ye-Kyung;Kim, Soon-Dong;Lee, In-Ae;Choi, Jongkeun;Suh, Joo-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1043-1053
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    • 2013
  • We investigated the ability of soybean curd residue (SCR) and its fermented products to inhibit obesity and improve the blood lipid profiles of obese mice fed a high-fat diet. Samples were prepared by fermenting SCR with Aspergillus oryzae var effuses KACC 44990 (ASCR), a microbe used for the fermentation of traditional Korean Meju, and with Monascus pilosus IFO 4480 (MSCR), a microbe used for the production of red rice. In addition, AMSCR, a mixture composed of equal amounts of ASCR and MSCR, was also prepared. Male mice were divided into six groups and fed with either a normal diet, a high-fat diet, or a high-fat diet supplemented with SCR, ASCR, MSCR, or AMSCR. After 8 weeks, body weight gain, serum and hepatic lipid profiles, and the activities of enzymes that generate or scavenge reactive oxygen species (ROS) were evaluated. Compared with the high-fat diet group, all the test groups showed a significant reduction in body, organ, and epididymal fat weight gain. These effects were observed with supplements in the order AMSCR>ASCR>MSCR>SCR. Similarly, supplements of test samples reduced high levels of serum and hepatic triglycerides (TG), total cholesterol, and low-density lipoprotein (LDL) cholesterol caused by hight-fat diet, while high-density lipoprotein (HDL) cholesterol was increased. Interestingly, the ability of ASCR to lower serum TG was stronger than that of MSCR, while MSCR showed a stronger hypocholesterolemic effect than ASCR. Meanwhile, AMSCR returned comprehensively serum lipid levels to normal. In addition, hepatic damage was prevented with effects in the order AMSCR>ASCR>MSCR>SCR. Hepatic ROS generating system including xanthine oxidase (XO) and ROS scavenging system including superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione S-transferase (GST) were recovered to normal level by all test diets. In conclusion, this study suggests that SCR and its fermented products can inhibit obesity and improve lipid profiles.

Anticarcinogenic Responses of MCF-7 Breast Cancer Cells to Conjugated Linoleic Acid (CLA) (식이성 Conjugated Linoleic Acid (CLA)가 유선암 세포(MCF-7)에서의 항암효과에 미치는 영향)

  • 문희정;이순재;박수정;장유진;이명숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.3
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    • pp.418-427
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    • 2003
  • Conjugated linoleic acid (CLA) is the mixture of positional and geometric isomers of linoleic acid (LA, C18:2 $\omega$6), which is found abundantly in dairy products and meats. This study was peformed to investigate the anticarcinogenic effect of CLA in MCF-7 breast cancer cells. MCF-7 cell were treated with LA and CLA at the various concentrations of 15, 30, 60, 120 UM each. After incubation for 48 and 72 hours, cell proliferation, fatty acids incorporation into cell, peroxidation and activities of antioxidant enzymes were measured. Postaglandin E$_2$ (PGE$_2$) and thromboxane $A_2$ (TXA$_2$) were measured for the eicosanoids metabolism. There was no cell growth differences in both of LA and CLA treated MCF-7 cells at 48 hr incubation. Compared to LA, cell growth was decreased by CLA treatment according to increasing concentration at longer incubation times, respectively (p<0.05). Both of LA and CLA was incorporated into the cellular lipids 22~54% higher than in control but LA incorporation was not so linear as CLA according to concentration. Arachidonic acid (C20:4, $\omega$6) was synthesized after treatment of LA but did not in CLA, respectively. The lipid peroxide concentration in LA 120 $\mu$M group increased as 1.7 times as that in CLA 120 $\mu$M treated. The activities of antioxidant enzymes such as glutathione peroxidase and glutathione reductase were increased by the supplementation with CLA 120 $\mu$M at 72 hr incubation (p<0.001) compared to LA, otherwise activity of superoxide dismutase was not different in both. PGE$_2$ and TXA$_2$ levels were lower in condition of CLA treatments according to lower levels of arachidonic acids than those in LA treated group, respectively. Overall, the dietary CLA might change the MCF-7 cell growth by the changes of cell composition, production of lipid peroxide, activities of antioxidant enzymes and eicosanoid synthesis compared to dietary LA.

Expression, Purification and Antiserum Production of the Avian Influenza H9N2 Virus HA and NA Proteins (Avian Influenza H9N2 Virus의 HA와 NA 단백질 발현, 정제 및 항혈청 생산)

  • Lee, Hyun-Ji;Song, Byung-Hak;Kim, Jeong-Min;Yun, Sang-Im;Kim, Jin-Kyoung;Kang, Young-Sik;Koo, Yong-Bum;Jeon, Ik-Soo;Byun, Sung-June;Lee, Youn-Jeong;Kwon, Jun-Hun;Park, Jong-Hyeon;Joo, Yi-Seok;Lee, Young-Min
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.178-185
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    • 2008
  • Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.

Fermentation Characteristics of Cheese Slurry prepared from Caseinates (카세인을 이용한 치즈곤죽의 발효특성)

  • Jang, Hae-Dong;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.17 no.5
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    • pp.389-398
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    • 1985
  • To shorten the processing of cheese slurry, four different slurries, ie, Control, Cheddar 1 and 2, and Italian-type that were made of Na-caseinates, cream, trace elements, lactic culture, and enzymes were fermented at $30^{\circ}C$ for 7days with daily stirring. PH, titratable acidity, soluble nitrogen, viable cell count, active SH groups, total volatile fatty acid, free fatty acid, electrophoretic patterns of degraded caseins, and viscosity were analyzed to investigate physicochemical properties of fermented slurries. Acid production was accelerated in the cheese slurries with protease than that without the enzyme and PH of the former was decreased after three days of fermentation to 4.90. The Change of titratable acidity agreed to PH patterns. Soluble nitrogen of the Control slurry was increased slowly for four days and then rapidly to 40% of total nitrogen while those containing protease to 70%. The protease of lactic cultures used (Streptococcus lactis and Streptococcus cremoris) broke down as-casein more rapidly than $\beta$-casein and most proteins were degraded to peptides and amino acids after three days of fermentation. Total volatile fatty acids were increased by added lipase and free fatty acids composition analyzed by GLC in cheddar slurry with 0.00001% lipase was similar to that of commercial cheddar cheese, while that in Italian-type slurry was a half of that in commercial Italian cheese. Active SH groups were increased in the cheese slurries with glutathione from fourth day of fermentation. The viscosity of slurries decreased very rapidly by addition of protease.

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Antioxidant Effects of Cysteine-containing Peptides of Different Lengths in Human HaCaT Keratinocytes Exposed to Hydrogen Peroxide (과산화수소에 노출된 인간 각질형성세포에서 길이가 다른 시스테인 함유 펩타이드의 항산화 효과)

  • Jae Won Ha;Joon Yong Choi;Yong Chool Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.193-201
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    • 2023
  • Hydrogen peroxide (H2O2) is a type of active oxygen species (ROS) that causes oxidative stress in cells and affects cell growth, proliferation, senescence, and death. The purpose of this study is to find active peptides that attenuate cytotoxicity of H2O2. A positional scanning synthetic tetrapeptide combinatorial library was screened to predict the sequence of potentially active peptides. As a result of comparing the effect of peptide pools on H2O2-induced death of human keratinocytes (HaCaT cells), various active peptide sequences were predicted. Especially, peptides containing cysteine (C) residue were predicted to be active. In follow-up experiments, the cytotoxicity and activity of cysteine-containing peptides of different lengths, such as C-NH2, CC-NH2, CCC-NH2, and CCCC-NH2 were examined. C-NH2 and CC-NH2 showed no significant cytotoxicity up to 1.0 mM, but CCC-NH2, and CCCC-NH2 showed relatively strong cytotoxicity. C-NH2 and CC-NH2 alleviated H2O2-induced cytotoxicity. CC-NH2 was more cytoprotective compared to C-NH2, C, N-acetyl cysteine (NAC), and glutathione (GSH). When intracellular ROS was measured by flow cytometry, H2O2 increased ROS production, and CC-NH2 suppressed ROS production more effectively than C-NH2, and it was as effective as C, NAC, and GSH. This study suggests that CC-NH2 of the cysteine-containing peptides of different lengths has an antioxidant property that safely and effectively alleviates H2O2-induced cytotoxicity and ROS production.

Effects of Dietary L-carnitine Supplementation on Growth Performance, Organ Weight, Biochemical Parameters and Ascites Susceptibility in Broilers Reared Under Low-temperature Environment

  • Wang, Y.W.;Ning, D.;Peng, Y.Z.;Guo, Y.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.2
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    • pp.233-240
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    • 2013
  • The objective of this study was to investigate the effects of L-carnitine on growth performance, organ weight, biochemical parameters of blood, heart and liver, and ascites susceptibility of broilers at different ages reared under a low-temperature environment. A total of 420 1-d-old male Ross 308 broilers were randomly assigned to two dietary treatments with fifteen replicates of fourteen broilers each. Treatment diets consisted of L-carnitine supplementation at levels of 0 and 100 mg/kg. At 11-d of age, low temperature stress was used to increase ascites susceptibility. Blood, heart and liver samples were collected at different ages for analysis of boichemical parameters. The results showed that, there was no significant difference in growth performance with L-carnitine supplementation, but the mortality due to ascites was significantly decreased. Dietary L-carnitine supplementation significantly reduced heart index (HI) and ascites heart index (AHI) on d 21, lung index (LUI) on d 35 and liver index (LI) on d 42. The broilers fed diets containing L-carnitine had significantly lower red blood cell counts (RBC), hemoglobin (HGB) concentration and hematocrit (HCT) on d 42. Dietary L-carnitine supplementation significantly reduced malondialdehyde (MDA) content of heart tissue on d 21 and 35, and significantly increased total superoxide dismutase (T-SOD) and Glutathione peroxidase (GSH-Px) activity of the heart on d 21 and 42. L-carnitine supplementation significantly reduced serum triglyceride (TG) content on d 28 and 35 and serum glucose (GLU) on d 35 and 42, and significantly increased serum total protein (TP) and globulin (GLO) content on d 42. L-carnitine supplementation significantly enhanced liver succinodehydrogenase (SDH), malic dehydrogenase (MDH) and $Na^+$-$K^+$-ATPase activity on d 28, and tended to reduce the lactic acid (LD) level of liver on d 35 (p = 0.06). L-carnitine supplementation significantly reduced serum uric acid (UA) content on d 28, 35 and 42. Based on the current results, it can be concluded that dietary L-carnitine supplementation reduced organ index, red blood cell counts and hematocrit, enhanced antioxidative capacity of the heart, enhanced liver enzymes activity involved in tricarboxylic acid cycle, and reduced serum glucose and triglyceride. Therefore, it is suggested that L-carnitine can potentially reduce susceptibility and mortality due to ascites.

A Novel Polyclonal Antiserum against Toxoplasma gondii Sodium Hydrogen Exchanger 1

  • Xiao, Bin;Kuang, Zhenzhan;Zhan, Yanli;Chen, Daxiang;Gao, Yang;Li, Ming;Luo, Shuhong;Hao, Wenbo
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.21-29
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    • 2016
  • The sodium hydrogen exchanger 1 (NHE1), which functions in maintaining the ratio of $Na^+$ and $H^+$ ions, is widely distributed in cell plasma membranes. It plays a prominent role in pH balancing, cell proliferation, differentiation, adhesion, and migration. However, its exact subcellular location and biological functions in Toxoplasma gondii are largely unclear. In this study, we cloned the C-terminal sequence of T. gondii NHE1 (TgNHE1) incorporating the C-terminal peptide of NHE1 (C-NHE1) into the pGEX4T-1 expression plasmid. The peptide sequence was predicted to have good antigenicity based on the information obtained from an immune epitope database. After induction of heterologous gene expression with isopropyl-b-D-thiogalactoside, the recombinant C-NHE1 protein successfully expressed in a soluble form was purified by glutathione sepharose beads as an immunogen for production of a rabbit polyclonal antiserum. The specificity of this antiserum was confirmed by western blotting and immunofluorescence. The antiserum could reduce T. gondii invasion into host cells, indicated by the decreased TgNHE1 expression in T. gondii parasites that were pre-incubated with antiserum in the process of cell entry. Furthermore, the antiserum reduced the virulence of T. gondii parasites to host cells in vitro, possibly by blocking the release of $Ca^{2+}$. In this regard, this antiserum has potential to be a valuable tool for further studies of TgNHE1.

Effect of 7,8-Dihydroxyflavone on In Vitro Maturation of Oocytes in Pigs

  • Oh, Seung-Kyu;Malaweera, Don Buddika Oshadi;Ramachandra, Sisitha;Shin, Sang-Tae;Cho, Jong-Ki
    • Journal of Embryo Transfer
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    • v.29 no.1
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    • pp.67-71
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    • 2014
  • In porcine embryo culture, one of reactive oxygen species (ROS) is harmful factors that are made during in vitro culture. To decrease the detrimental effect of ROS on embryo development, superoxide dismutase, catalase and glutathione peroxidase could be used in the embryo culture. Out of these antioxidants, 7,8-dihydroxyflavone (7,8-DHF) was reported its antioxidant effects to prevent the glutamine-triggered apoptosis. Therefore, this study was performed to investigate the most appropriate concentration of 7,8-DHF in porcine embryonic development. For that, 5 different concentration (0, 0.1, 0.5, 1, $2{\mu}m$) of 7,8-DHF was supplemented in the porcine IVM media and then maturation and blastocyst formation rates were compared among 5 groups. In maturation rates of porcine oocytes, significant higher maturation rates was shown in the $1.0{\mu}m$ group compared with another 4 groups ($83.3{\pm}2.1$ vs. $80.7{\pm}1.4$, $79.8{\pm}1.4$, $78.3{\pm}1.2$, $79.4{\pm}1.6$), respectively (P<0.05). In the embryo culture, $1.0{\mu}m$ group also showed the significant higher cleavage rates ($76.8{\pm}3.1$ vs. $62.1{\pm}5.0$, $65.7{\pm}4.0$, $68.6{\pm}3.7$, $64.6{\pm}4.0%$) and blastocyst formation rates - ($39.6{\pm}4.0%$ vs. $28.6{\pm}3.3$, $31.1{\pm}3.9$, $29.3{\pm}2.5$, $39.6{\pm}4.0$, $26.4{\pm}3.2%$), respectively (P<0.05). There was no significant difference among 5 groups in the cell number of blastocyst (P<0.05). In conclusion, supplement of $1.0{\mu}m$ of 7,8-DHF was effective to increase the porcine embryonic development competence as antioxidant to ROS.