• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.2
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• pp.99-106
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• 2011

In order to investigate rice stem proteome in response to heat stress, rice plants were subjected to heat treatment at 42$^{\circ}C$ and total soluble proteins were extracted from stem tissues, and were fractionated with 15% PEG (poly ethylene glycol) and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). After staining of 2-DE gels, 46 of differentially expressed proteins were extracted, digested by trypsin, and subjected to matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Proteins were identified through database search by using peptide mass fingerprints. Among them, 10 proteins were successfully identified. Seven proteins were up- and 3 proteins were down-regulated, respectively. These proteins are involved in energy and metabolism, redox homeostasis, and mitochondrial small heat shock proteins. The identification of some novel proteins in the heat stress response provides new insights that can lead to a better understanding of the molecular basis of heat-sensitivity in plants, and also useful to molecular breeding of thermotolerant forage crops.

• Tuberculosis and Respiratory Diseases
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• v.59 no.1
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• pp.30-38
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• 2005

Background : Arsenic trioxide ($As_2O_3$) has been used to treat acute promyelocytic leukemia, and it induces apoptosis in a variety of solid tumor cell lines including non-small cell lung cancer cells. However, nonsteroidal antiinflammatory drugs (NSAID) can enhance tumor response to chemotherapeutic drugs or radiation. It was previously demonstrated that a combination treatment with $As_2O_3$ and sulindac induces the apoptosis of NCI-H157 human lung carcinoma cells by activating the caspase cascade. This study aimed to determine if a combination treatment augmented its apoptotic potential through other pathways except for the activation of the caspase cascade. Material and Methods : The NCI-H157 cells were treated with $As_2O_3$, sulindac and antioxidants such as glutathione (GSH) and N-acetylcysteine (NAC). The cell viability was measured by a MTT assay, and the level of intracellular hydrogen peroxide ($H_2O_2$) generation was monitored fluorimetrically using a scopoletin-horse radish peroxidase (HRP) assay. Western blotting and mitochondrial membrane potential transition analysis were performed in order to define the mechanical basis of apoptosis. Results : The viability of the cells was decreased by a combination treatment of $As_2O_3$ and sulindac, and the cells were protected using antioxidants in a dose-dependent manner. The increased $H_2O_2$ generation by the combination treatment was inhibited by antioxidants. The combination treatment induced changes in the mitochondrial transmembrane potential as well as the expression of the Bcl-2 family proteins, and increased cytochrome c release into the cytosol. However, the antioxidants inhibited the effects of the combination treatment. Conclusion : Combination treatment with $As_2O_3$ and sulindac induces apoptosis in NCI-H157 human lung carcinoma cells via ROS generation with a mitochondrial dysfunction.

• The Korean Journal of Zoology
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• v.31 no.1
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• pp.62-70
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• 1988

The distribution of respiratory chain complexes in beef heart and human muscle mitochondria has been explored by immunoeledron microscopy with antibodies made against beef heart mltochondriai proteins in conjundion with protein A cofloidai gold (l2nm particles). The antibodies used were made against NADH-conezyme Q reductase(complex I), ubiquinol-cytochrome-c-oxldoreductase (complex III) and cytochrome-c-oxidase(complex IV). Labeling of bed heart tissue with any of these antihodies gave gold particles randomly distributed along the mitochondrial inner membrane. The labeling of muscle tIssue mitochondria from a patient with a mitochondrial myopathy localized by biochemical analysis to complex III was quantitated and compared with the labeling of human control muscle tissue mitochondria. Four kinds of morphological changes in the mitochondrial fine strudure in the myopathy patient tissue have been found; paracrystalline inclusions consistIng of densely packed multi- lamellar structures, globular crystalline inclusions with high electron density, multilamellar strudure inclusion body(compadly and irregularly arranged concentric whirl shaped cristae)and golbular cyrstalilne inclusions located in the center of the whirl shaped cristae. Compex I and cytochrome-c-oxldase antihodies reacted to the same level in the mitochondria containing the crystalline inclusions and control mitochondria. Antibodies to complex III reacted very poorly to the mitochondria containing the crystalline Inclusions but strongly to control mitchondria. The globular crystalline inclusions in the mitochondria are not reacted antibodies to respiratory chain complexes.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.1
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• pp.78-85
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• 2019

Membrane proteins are involved in many common diseases, including heart disease and cancer. In various disease states, such as cancer, abnormal signaling pathways that are related to the membrane proteins cause the cells to divide out of control and the expression of membrane proteins can be altered. Membrane proteins have the hydrophobic environment of a lipid bilayer, which makes an analysis of the membrane proteins notoriously difficult. Therefore, this study evaluated the efficacy of two different methods for optimal membrane protein extraction. High-speed centrifuge and reagent-based method with a -/+ filter aided sample preparation (FASP) were compared. As a result, the high-speed centrifuge method is quite effective in analyzing the mitochondrial inner membranes, while the reagent-based method is useful for endoplasmic reticulum membrane analysis. In addition, the function of the membrane proteins extracted from the two methods were analyzed using GeneGo software. GO processes showed that the endoplasmic reticulum-related responses had higher significance in the reagent-based method. An analysis of the process networks showed that one cluster in the high-speed centrifuge method and four clusters in the reagent-based method were visualized. In conclusion, the two methods are useful for the analysis of different subcellular membrane proteins, and are expected to assist in selecting the membrane protein extraction method by considering the target subcellular membrane proteins for study.

• Journal of Nutrition and Health
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• v.56 no.6
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• pp.602-614
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• 2023

Purpose: The balance between synthesis and degradation of proteins plays a critical role in the maintenance of skeletal muscle mass. Mitochondrial dysfunction has been closely associated with skeletal muscle atrophy caused by aging, cancer, and chemotherapy. Polygalacin D is a saponin derivative isolated from Platycodon grandiflorum (Jacq.) A. DC. This study aimed to investigate the effects of polygalacin D on myoblast differentiation and muscle atrophy in association with mitochondrial function in in vitro and in zebrafish models in vivo. Methods: C2C12 myoblasts were cultured in differentiation media containing different concentrations of polygalacin D, followed by the immunostaining of the myotubes with myosin heavy chain (MHC). The mRNA expression of markers related to myogenesis, muscle atrophy, and mitochondrial function was determined by real-time quantitative reverse transcription polymerase chain reaction. Wild type AB^* zebrafish (Danio rerio) embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without polygalacin D, and immunostained to detect slow and fast types of muscle fibers. The Tg(Xla.Eef1a1:mitoEGFP) zebrafish expressing mitochondria-targeted green fluorescent protein was used to monitor mitochondrial morphology. Results: The exposure of C2C12 myotubes to 0.1 ng/mL of polygalacin D increased the formation of MHC-positive multinucleated myotubes (≥ 8 nuclei) compared with the control. Polygalacin D significantly increased the expression of MHC isoforms (Myh1, Myh2, Myh4, and Myh7) involved in myoblast differentiation while it decreased the expression of atrophic markers including muscle RING-finger protein-1 (MuRF1), mothers against decapentaplegic homolog (Smad)2, and Smad3. In addition, polygalacin D promoted peroxisome proliferator-activated receptor-gamma coactivator (Pgc1α) expression and reduced the level of mitochondrial fission regulators such as dynamin-1-like protein (Drp1) and mitochondrial fission 1 (Fis1). In a zebrafish model of FOLFIRI-induced muscle atrophy, polygalacin D improved not only mitochondrial dysfunction but also slow and fast muscle fiber atrophy. Conclusion: These results demonstrated that polygalacin D promotes myogenesis and alleviates chemotherapy-induced muscle atrophy by improving mitochondrial function. Thus, polygalacin D could be useful as nutrition support to prevent and ameliorate muscle wasting and weakness.

• Journal of the Korean Applied Science and Technology
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• v.37 no.4
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• pp.744-754
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• 2020

We investigated the whether endurance exercise and MitoQ intake mediated neuroprotection are associated with mitochondrial function in 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine(MPTP) -induced mice model of Parkinson's disease. C57BL/6 male mice were randomly assigned to five groups: Normal Conrol(NC, n=10), MPTP Control(MC, n=10), MPTP +MitoQ(MQ, n=10), MPTP + Exercise(ME, n=10) and MPTP + MitoQ + Exercise(MQE, n=10). Exercise intervention groups performed the treadmill exercise for 5days/week for 5 weeks with gradual increase of intensity. MitoQ intake groups consumed the MitoQ at a concentration of 250μmol by dissolving with water during experiment period. Our data demonstrated that ME and MQE group restored MPTP-induced motor dysfunction. In addition, treatment groups(MQ, ME and MQE) increased tyrosine hydroxylase levels, and suppressed the accumulation of α-synuclein levels. Futhermore, treatment groups modulated the mitochondrial function such as upregulated mitochondrial biogenesis, increased antioxidant enzyme, enhanced a anti-apoptotic protein(e.g., BCL2), and reduced a pro-apoptotic protein(e.g., BAX). Taken together, these results suggested that endurance exercise and MitoQ intake-mediated increase in mitochondrial function contributes to improvement of aggravated dopaminergic neuronal, resulting in attenuation of motor function of Parkinson's disease.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.4
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• pp.682-688
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• 1997

This study was performed to investigate whether ascorbic acid can modulate the induction of CYP2E1 and prevent the lipid peroxidation which may cause diabetic chronic complication. Diabetes was induced by intraperitoneal injection of streptozotocin to 5-week-old male Sprague-Dawley rats(150~170g). Normal and diabetic group was randomly divided into three groups each; Control(CON, no supplementation), SUP1 (50mg/d ascorbate supplementation) and SUP2(250mg/d ascorbate supplementation). Ascobic acid was prepared daily by dissolving in drinking water and supplied for 4 weeks. There was no difference in hepatic microsomal and mitochondrial P450 contents between normal and diabetes. Hepatic microsomal N-nitrosodimethylamine(NDMA) demethylase activity, which repre-sents contents of CYP2E1, was elevated in diabetes, but not significantly. The NDMA demethylase activity of diabetic SUP2 group was significantly lower activity than that of the diabetic CON group. However, no difference in hepatic mitochondrial NDMA demethylase activity was observed between the diabetes and the normal group. The result suggests that the induction of CYP2E1 in diabetes can be alleviated by ascorbic acid supplementation at the dose of 50mg1d. In addition, ascorbic acid supplementation showed dose-dependent reduction of hepatic microsomal TBARS contents in diabetic rats.

• Bulletin of Food Technology
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• v.23 no.2
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• pp.214-219
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• 2010

Saccharomyces cerevisiae는 전통적으로 알코올 음료와 bioethanol 생산에 이용되지만, 발효가 진행되는 동안 효모의 에탄올 생성은 에탄올의 축적에 의한 충격으로 세포활성에 손상을 초래한다. 본 연구는 S. cerevisiae의 에탄올 스트레스 반응과 에탄올 내성의 분자적 기초에 관해 수행되었으며, 에탄올 스트레스가 진행되는 동안 효모의 에탄올 생성 향상을 위한 유전 공학 전략의 수립에 활용될 수 있다. 이전의 연구들은 유전자 발현에 대한 에탄올 스트레스의 충격이 환경적 영향을 받기 때문에 다양한 균주와 조건들에 관해 이루어졌다. 그러나 에탄올 공격에 의해 영향을 받은 gene ontology 범주에서의 일부 공통점은 S. cerevisiae의 에탄올 스트레스 반응이 해당과정 및 미토콘드리아 기능과 관련된 유전자 발현의 증가와 에너지가 요구되는 성장과정과 관련된 유전자의 발현 감소에 따라 에너지 생산에 제약 받음을 의미한다. Genomewide screens를 이용한 연구는 vacuole function의 유지가 에탄올 내성에 대해 중요함을 암시한다. 아마도 단백질 turnover와 이온 항상성 유지에 이 세포기관의 역할이 중요하기 때문인 것으로 사료된다. 특히 에탄올 스트레스가 일어날 때 핵 내 Asr1과 Rat8의 축적은 비록 이 가설이 논란이 많은 주제로 남아있지만 S. cerevisiae가 에탄올 스트레스에 대한 특별한 반응을 가지고 있음을 의미한다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.9
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• pp.1257-1264
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• 2016

Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.08a
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• pp.36-37
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• 2001

큰가리비는 자웅이체이다. 난환형성과정은 난모세포의 발달정도에 다라 다르게 나타나고 있다. 난자형성과정은 난원세포기, 전난황형성난모세포기, 초기난황형성난모세포기, 후기난황형성난모세포기, 성숙난모세포기의 연속적인 5단계의 과정으로 나눌 수 있었다. 전난황형성기 난모세포질 내에서는 핵주변 구역에 골지장치와 수많은 공포들 및 미토콘드리아들이 출현하고 있는데 이들은 차후, 지방적 형성에 관여한다. 난황형성전기난모세포(previtellogenic oocyte)에서는 지방적 및 지질과립들이 핵막 근처에서 출현하여 피질층쪽으로 분산되는 반면, 같은 발달 단계의 난모세포질의 피질구역에서는 피질과립들(단백질성 난황과립)이 처음으로 생성되어 난황막 근처의 피질층에서 핵주변 구역쪽으로 분산.분포된다. 난황형성후기 난모세포에서는 세포질 내의 골지장치, 공포, 미토콘드리아, 그리고 조면소포체들이 자율합성에 의해 난황과립 형성에 관여하고 있다. 반면 외인성 물질들인 지질형태의 과립들, 다량의 글리코겐 입자들이 생식상피 내에서 출현하고 있는데. 이들 물질이 생식상피에서 난황막 구조물인 미세융모를 통해 난황형성 후기 난모세포의 날질 내로 통과해 들어가는 현상이 관찰되었다. 이와같은 현상은 난황형성이 일어날 때에 heterosynthesis가 일어나고 있음을 시사한다. 완숙난모세포의 난경은 약 50~60$\mu\textrm{m}$이다. 정자형성과정은 정원세포기, 제1차정모세포기, 제2차정모세포기, 정세포기, 정자기의 연속적인 5단계로 나눌수 있었다. 정셍포기에서 정자로 변태되는 과정 중에 침체의 분화과정이 있는데 이에는 1. Golgi phase, 2. Cap phase 3. acrosome phase, 4. maturation phase의 단계를 거쳐 첨체가 완성된다. 정자는 원시적 형태를 이루고 있으며 4개의 미토콘드리아가 부핵을 형성하고 있다. 완숙정자 두부의 길이는 대략 $3 \mu$m 이며, 미부의 길이는 약 $30 \mu$m정도이다. 정자 미부편모의 axoneme은 중앙의 2개의 미세소관(microtubule)과 주변에 위치한 9개의 2중 미세소구관(microtublue)으로 이루어져 있다.