• Asian-Australasian Journal of Animal Sciences
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• 제33권3호
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• pp.424-435
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• 2020

Objective: The study was conducted to investigate variations in the immunophysiological responses to exercise-induced stress in Jeju and Thoroughbred horses. Methods: Blood samples were collected from the jugular veins of adult Jeju (n = 5) and Thoroughbred (n = 5) horses before and after 30 min of exercise. The hematological, biochemical, and immunological profiles of the blood samples were analyzed. Blood smears were stained and observed under a microscope. The concentration of cell-free (cf) DNA in the plasma was determined using real time polymerase chain reaction (PCR). Peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells were separated using Polymorphprep, and the expression of various stress-related and chemokine receptor genes was measured using reverse transcriptase (RT) and real-time PCR. Results: After exercise, Jeju and Thoroughbred horses displayed stress responses with significantly increased rectal temperatures, cortisol levels, and muscle catabolism-associated metabolites. Red blood cell indices were significantly higher in Thoroughbred horses than in Jeju horses after exercise. In addition, exercise-induced stress triggered the formation of neutrophil extracellular traps (NETs) and reduced platelet counts in Jeju horses but not in Thoroughbred horses. Heat shock protein 72 and heat shock protein family A (Hsp70) member 6 expression is rapidly modulated in response to exercise-induced stress in the PBMCs of Jeju horses. The expression of CXC chemokine receptor 4 in PBMCs was higher in Thoroughbred horses than in Jeju horses after exercise. Conclusion: In summary, the different immunophysiological responses of Jeju and Thoroughbred horses explain the differences in the physiological and anatomical properties of the two breeds. The physiology of Thoroughbred horses makes them suitable for racing as they are less sensitive to exercise-induced stress compared to that of Jeju horses. This study provides a basis for investigating the link between exercise-induced stresses and the physiological alteration of horses. Hence, our findings show that some of assessed parameters could be used to determine the endurance performance of horses.

• BMB Reports
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• 제38권2호
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• pp.151-160
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• 2005

By a cDNA array representing 2308 signal transduction related genes, we studied the expression profiles of HeLa cells stably transfected by Hepatitis C virus nonstructural protein 4B (HCV-NS4B). The alterations of the expression of four genes were confirmed by real-time quantitative RT-PCR; and the aldo-keto reductase family 1, member C1 (AKR1C1) enzyme activity was detected in HCV-NS4B transiently transfected HeLa cells and Huh-7, a human hepatoma cell line. Of the 2,308 genes we examined, 34 were up-regulated and 56 were down-regulated. These 90 genes involved oncogenes, tumor suppressors, cell receptors, complements, adhesions, transcription and translation, cytoskeletion and cellular stress. The expression profiling suggested that multiple regulatory pathways were affected by HCV-NS4B directly or indirectly. And since these genes are related to carcinogenesis, host defense system and cell homeostatic mechanism, we can conclude that HCV-NS4B could play some important roles in the pathogenesis mechanism of HCV.

• 대한본초학회지
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• 제32권3호
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• pp.55-61
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• 2017

Objectives : The aim of this research was to determine the diverse effects of Lithospermi Radix Water Extract (LR) on human keratinocyte HaCaT cells, and to examine whether those effects could be applied to the human skin. Methods : We examined effect of LR on the cell viability of using the MTS assay in human keratinocyte HaCaT cells. The antioxidation effect of LR was analyzed relative to the well-known antioxidant resveratrol, using an ABTS assay. Quantitative RT-PCR analysis revealed that, in HaCaT cells, LR influenced the mRNA expression of tight-junction genes associated with skin moisturization. Furthermore, a wound-healing assay demonstrated altered cell migration in LR-treated HaCaT cells. Result : The cytotoxicity was confirmed to be higher in LR at a concentration of $800{\mu}g/m{\ell}$ using the MTS assay in HaCaT cells. In comparison to $100{\mu}M$ resveratrol, $1,600{\mu}g/m{\ell}$ LR showed either a similar or superior antioxidation effect. LR treatment in HaCaT cells reduced the mRNA expression levels of claudin 3, claudin 4, claudin 6, claudin 8, and ZO-2 to less than 0.80-fold, whereas JAM-A and Tricellulin mRNA expression level increased more than 1.33-fold. In addition, HaCaT cells migration was decreased to 83.9% by LR treatment. Conclusions : LR of antioxidation activity will have an anti-aging effect on the skin by reducing oxidative stress. Further studies are required to address the implications for human skin, given LR's effects of altering mRNA expression of tight junction-related gene and decreasing cell migration of HaCaT cells.

• 동의생리병리학회지
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• 제25권4호
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• pp.663-668
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• 2011

Galgeun-tang (GGT) has been a great source for treating cold diseases in the folk medicine recipe. Carbon tetrachloride ($CCl_4$) is one type of hepatotoxin that can eventually cause liver injury. During the experiment, we first studied the protective effects of GGT against $CC_4$-induced hepatotoxicity. GGT was pretreated for 3 h, and 1% $CCl_4$ was added to mouse primary liver cells. After 4 h, ROS generation and expression of antioxidant enzymes (catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx)) were analyzed by FACS and real time PCR. Also, the activities of ALT and LDH were measured using cultured medium. The hepatic levels of TNF-alpha and iNOS, which are related to inflammation and stress response gene, HSP72 and HO-1 were analyzed by PCR or real time PCR. Liver tissues were analyzed by HE stain. From the observation, we discovered that GGT treatment protects $CCl_4$-induced hepatotoxicity, and that GGT pretreatment decreases ROS generation, TNF-alpha and iNOS expression. However, gene expression of CAT, SOD, GPx, HSP72 and HO-1 were increased by GGT. These results lead to the conclusion that GGT has protective effects against $CCl_4$-induced hepatotoxicity.

• 한국해양생명과학회지
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• 제3권2호
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• pp.59-66
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• 2018

갯벌의 건강 수준에 대한 평가는 갯벌에 서식하는 생물의 건강에 의해 평가될 수 있다. 생체지표유전자의 발현 분석을 통하여 갯벌에 서식하는 생물의 건강 수준을 평가할 수 있다. 본 연구의 목적은 heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), glutathione S-transferases (GST) 및 thioredoxin (TRX)과 같은 생체지표유전자를 이용하여 서해안 갯벌의 건강을 평가하는 것이다. 이들 유전자는 스트레스, 면역 및 항산화 관련한 유전자들로서 이들 유전자의 발현을 통해 생물의 건강 정도를 관찰하는 데 사용할 수 있다. 본 연구에서는 서해안의 8개 정점에서 바지락(Ruditapes philippinarum)을 수집했다. 유전자의 발현은 RT-qPCR 방법으로 분석하였다. 연구 결과 Hsp70, Hsp90, GST 및 TRX 유전자들의 발현이 8개 정점에서 차별적으로 발현되는 것으로 나타났다. 특히, Hsp90 및 GST의 발현 또는 Hsp70 및 TRX의 발현은 유사하였다. 이것은 각 유전자에 특이적으로 반응하는 물질이 존재하는 것으로 생각된다. 따라서 이화학적 분석결과에 근거하여 분석에 적합한 유전자를 선택할 수 있다고 생각한다. 이 결과는 Hsp70, Hsp90, GST 및 TRX 유전자는 갯벌의 건강을 평가하기 위한 생체지표유전자로서의 역할을 수행함을 시사한다.

• Biomolecules & Therapeutics
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• 제29권1호
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• pp.90-97
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• 2021

Ultraviolet B (UVB) radiation causes DNA base modifications. One of these changes leads to the generation of 8-oxoguanine (8-oxoG) due to oxidative stress. In human skin, this modification may induce sunburn, inflammation, and aging and may ultimately result in cancer. We investigated whether phloroglucinol (1,3,5-trihydroxybenzene), by enhancing the expression and activity of 8-oxoG DNA glycosylase 1 (Ogg1), had an effect on the capacity of UVB-exposed human HaCaT keratinocytes to repair oxidative DNA damage. Here, the effects of phloroglucinol were investigated using a luciferase activity assay, reverse transcription-polymerase chain reactions, western blot analysis, and a chromatin immunoprecipitation assay. Phloroglucinol restored Ogg1 activity and decreased the formation of 8-oxoG in UVB-exposed cells. Moreover, phloroglucinol increased Ogg1 transcription and protein expression, counteracting the UVB-induced reduction in Ogg1 levels. Phloroglucinol also enhanced the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) as well as Nrf2 binding to an antioxidant response element located in the Ogg1 gene promoter. UVB exposure inhibited the phosphorylation of protein kinase B (PKB or Akt) and extracellular signal-regulated kinase (Erk), two major enzymes involved in cell protection against oxidative stress, regulating the activity of Nrf2. Akt and Erk phosphorylation was restored by phloroglucinol in the UVB-exposed keratinocytes. These results indicated that phloroglucinol attenuated UVB-induced 8-oxoG formation in keratinocytes via an Akt/Erk-dependent, Nrf2/Ogg1-mediated signaling pathway.

• Journal of Microbiology and Biotechnology
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• 제21권8호
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• pp.830-837
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• 2011

A genomic DNA fragment encoding a putative maltooligosyltrehalose trehalohydrolase (NfMTH) for trehalose biosynthesis was cloned by the degenerate primer- PCR from cyanobacterium Nostoc flagelliforme. The ORF of NfMTH is 1,848 bp in length and encodes 615 amino acid residues, constituting a 70 kDa protein. The deduced amino acid sequence of NfMTH contains 4 regions highly conserved for MTHs. By expression of NfMTH in E. coli, the function of this protein was demonstrated, where the recombinant protein catalyzed the hydrolysis of maltooligosyl trehalose to trehalose. The expressions of MTH and maltooligosyltrehalose synthase in the filaments of N. flagelliforme were upregulated significantly under dehydration stress, NaCl stress, and high temperature-drought stress. The accumulations of both trehalose and sucrose in the filaments of N. flagelliforme were also improved significantly under the above stresses. Furthermore, trehalose accumulated in smaller quantities than sucrose did when under NaCl stress, but accumulated in higher quantities than sucrose did when under temperature-drought stress, indicating that both trehalose and sucrose were involved in N. flagelliforme adapted to stresses and different strategies conducted in response to various stress conditions.

• 미생물학회지
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• 제46권2호
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• pp.113-121
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• 2010

인산 결핍기에 직면한 Bacillus subtilis는 PhoP-PhoR twocomponent system (TCS)를 통해 이러한 상황을 인식하고 생존을 유지하기 위해 Pho regulon으로 불리는 일련의 유전자들의 발현을 조절한다. 이때 histidine kinase인 PhoR은 자동 인산화되어, 인산을 response regulator인 PhoP에 전달한다. 인산화된 PhoP (PhoP~P)는 Pho regulon 유전자의 프로모터(promoter) 부위에 존재하는 반복되는 6 bp의 잘 보존된 PhoP 결합서열에 결합하여 해당 유전자의 발현을 활성화시키거나 억제한다. 이러한 Pho regulon 신호전달 시스템은 최소한 세 개의 TCS (PhoP-PhoR, ResD-ResE TCS, SpoOA phosphorelay), 광범위한 탄소대사 조절자(CcpA), 전위기 조절자(AbrB, ScoC) 등을 포함하는 신호전달 시스템과 밀접하게 상호 연결되어 있을 뿐만 아니라, 생육에 필수적인 YycF-YycG TCS와 상호조절을 통한 밀접한 관련을 가지고 있다. Pho regulon에 의한 인산결핍 스트레스 반응을 이해하는데 많은 진척이 있었으나, 많은 의문들은 여전히 남아있다. 이러한 의문들을 푸는 일은 B.subtilis의 응용연구에 중요한 정보를 제공할 것이다.

• Journal of Plant Biotechnology
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• 제43권2호
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• pp.204-212
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• 2016

환경스트레스 중의 하나인 저온에 대한 생육기의 포도나무의 반응을 분석하고자 -$2^{\circ}C$에서 4일 동안 저온처리 한두 품종('Campbell Early'와 'Muscat Baily A')의 포도나무잎을 이용하여 전사체를 분석하였고 특이발현유전자(differentially expressed genes, DEGs)를 검색하였다. 영하의 저온에 반응한 'Campbell Early'의 DEG를 기능별로 분석한 결과 생물대사에서 17,424개, 세포구성에서 28,954개, 분자기능에서는 6,972개의 유전자와 관련이 있었다. 발현이 유도되는 유전자로는 dehydrin xero 1, K-box region and MADS-box transcription factor family protein과 MYB domain protein 36이 있으며, 억제되는 유전자로는 light-harvesting chlorophyll B-binding protein 3, FASCICLIN-like arabinoogalactan 9와 pectin methylesterase 61 등이 있었다. 'Muscat Baily A'의 DEG는 생물대사에서 1,157개, 세포구성에서 1,350개, 분자기능에서는 431개의 유전자와 관련이 있었다. 발현이 유도되는 유전자로는 NB-ARC domain-containing disease resistance protein, fatty acid hydrozylase syperfamily와 isopentenyltransferase 3이 있으며, 억제되는 유전자로는 binding, IAP-like protein 1과 pentatricopeptide repeat superfamily protein 등이 있었다. Real-time PCR을 이용하여 영하의 저온에서 특이적으로 발현하는 유전자들을 검정하였으며, InterPro Scan을 통해 단백질 도메인을 분석한 결과 두 품종 모두에서 ubiquitin-protein ligase가 가장 많았다. 영하의 저온에 노출된 신초의 전사체 정보를 바탕으로 포도나무에서 저온 내성을 발현하는 기작을 연하는 데에 분자수준의 정보를 제공하고, 내한성 포도를 육종하는데 이용될 수 있을 것이다.

• Journal of Plant Biotechnology
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• 제35권2호
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• pp.133-140
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• 2008

DNA와 histone 단백질의 변형은 식물 발달에 상당히 중요한 역할을 하는 것으로 알려져 있다. 식물 조직 배양 및 식물 발달 단계에서 methylation의 영향을 알아보고자 벼 종자로부터 캘러스 형성 및 식물체 재분화 단계에서 demethylation 물질인 5-azacytidine을 처리하여 유전자 발현 양상을 분석하였다. 식물체로의 재분화 능력이 있는 벼 배상체 캘러스는 5-azaC가 첨가된 H6A 배지에서는 형성되지 않았으며 갈색을 띠는 캘러스가 형성되었다. 또한 정상적인 캘러스를 5-azaC가 첨가된 MSRA 재분화 배지에서 배양했을 때도 대조구와는 달리 식물체 재분화는 이루어지지 않았다. 이러한 결과는 5-azaC가 정상적인 배상체 캘러스 및 shoot 분화에 부정적인 영향을 미친다는 것을 나타냈으며 따라서 DNA methylation이 식물 조직배양에서의 정상적인 세포 dedifferentiation과 differentiation에 필수 요인이라는 것을 알 수 있었다. 벼 캘러스 형성 및 재분화 과정 동안의 methylation 영향을 알아보고자 각 단계별로 5-azaC를 처리 후 $GeneFishig^{TM}$ DEG와 DNA chip을 사용하여 유전자 발현 양상을 분석하였다. Epigenetic regulation, 전자전달, 핵산대사, 스트레스 반응에 관여하는 일부 유전자들의 발현이 증가하거나 감소하는 것을 알 수 있었다. 발현 차이가 있는 일부 유전자를 클로닝하여 확인하였고 RT-PCR 및 northern 분석으로 각 단계에서의 발현 차이를 할인하였다.