• Animal Bioscience
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• 제34권7호
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• pp.1210-1220
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• 2021

Objective: Unlike mammals, goose fatty liver shows a strong tolerance to fatty acids without obvious injury. Stearyl-coenzyme A desaturase 1 (SCD1) serves crucial role in desaturation of saturated fatty acids (SAFs), but its role in the SAFs tolerance of goose hepatocytes has not been reported. This study was conducted to explore the role of SCD1 in regulating palmitic acid (PA) tolerance of goose primary hepatocytes. Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide was examined to reflect the effect of PA on hepatocytes viability, and quantitative polymerase chain reaction was used to detect the mRNA levels of several genes related to endoplasmic reticulum (ER) stress, inflammation, and apoptosis, and the role of SCD1 in PA tolerance of goose hepatocytes was explored using RNA interfere. Results: Our results indicated that goose hepatocytes exhibited a higher tolerant capacity to PA than human hepatic cell line (LO2 cells). In goose primary hepatocytes, the mRNA levels of fatty acid desaturation-related genes (SCD1 and fatty acid desaturase 2) and fatty acid elongate enzyme-related gene (elongase of very long chain fatty acids 6) were significantly upregulated with 0.6 mM PA treatment. However, in LO2 cells, expression of ER stress-related genes (x box-binding protein, binding immunoglobulin protein, and activating transcription factor 6), inflammatory response-related genes (interleukin-6 [IL-6], interleukin-1β [IL-1β], and interferon-γ) and apoptosis-related genes (bcl-2-associated X protein, b-cell lymphoma 2, Caspase-3, and Caspase-9) was significantly enhanced with 0.6 mM PA treatment. Additionally, small interfering RNA (siRNA) mediated downregulation of SCD1 significantly reduced the PA tolerance of goose primary hepatocytes under the treatment of 0.6 mM PA; meanwhile, the mRNA levels of inflammatory-related genes (IL-6 and IL-1β) and several key genes involved in the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), forkhead box O1 (FoxO1), mammalian target of rapamycin and AMPK pathways (AKT1, AKT2, FoxO1, and sirtuin 1), as well as the protein expression of cytochrome C and the apoptosis rate were upregulated. Conclusion: In conclusion, our data suggested that SCD1 was involved in enhancing the PA tolerance of goose primary hepatocytes by regulating inflammation- and apoptosis-related genes expression.

• Clinical and Experimental Pediatrics
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• 제52권12호
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• pp.1348-1357
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• 2009

목 적:소아 전염성 단핵구증 환자의 임상적 특성과 EBV 감염과 관련된 사이토카인과 세포자멸사 연관 유전자의 발현에 대해 알아보고자 본 연구를 시행하였다. 방 법:2006년 3월부터 2007년 2월까지 전북대학교병원에 입원한 전형적인 임상 증상을 보이면서 혈청학적 검사로 전염성 단핵구증으로 확진된 15명을 대상으로 임상적 특성을 조사하고, 진단 당시, 발병 1주 후, 3주 후에 각각 말초혈액에서의 T세포 세군, 사이토카인, EBV DNA, 세포자멸사 연관 유전자 발현을 측정하여 건강한 대조군 10명과 비교하였다. 결 과:환자의 평균 연령은 $5.7{\pm}3.4$세(3-9세)였으며, 남녀비는 1.5:1이었다. 발열(100%)과 인두통(73%), 무기력과 권태감(53%)이 흔한 임상 증상이었고, 인두염 및 편도염(100%), 경부 림프절 비대(87%)와 간비종대(53%)가 흔하게 나타났다. 환자의 말초혈액에서 CD3+ T세포와 CD8+ suppressor T세포, CD56+ NK세포가 대조군에 비하여 증가하였다(P<0.01). 환자의 혈청 IL-2, 6, $INF-{\gamma}$가 증상 초기에 대조군에 비하여 증가하였다(P<0.01). 환자의 말초혈액 단핵구내 EBV DNA 양은 평균 $10^{2.38}copies/{\mu}g$이었으며 진단 당시 최고치를 보이다가 회복기에는 정상화되는 양상이었다. 환자의 말초혈액 단핵구의 세포자멸사 연관 유전자 발현은 Bcl-2는 증상 초기에 증가하다가 감소하였고 Bax는 회복기에 약간 증가하였으며, Fas는 거의 변화가 없었으나 FasL은 발병 1주 후에 현저히 증가하였다가 회복기에 감소하였다. 결 론:본 연구를 통해 급성 전염성 단핵구증 소아 환자에서 다양한 임상 증상과 말초혈액에서 T세포 세군의 변화 및 세포자멸사 연관 유전자 발현의 변화를 확인하였다. 앞으로 분자 생물학적 관점에서의 병인 및 치료 방법에 대한 더 많은 연구가 필요할 것으로 사료된다.

• 생명과학회지
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• 제34권7호
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• pp.500-508
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• 2024

최근 현대인들의 다중이용시설 사용 빈도가 증가하면서 실내 공기 질, 특히 실내공기 오염원인 공기매개 전염성 세균 및 바이러스 불안에 대한 관심이 높아져 다양한 형태의 공기청정기와 공기살균기의 보급이 확산되고 있다. 따라서 공기 내 미생물을 살균할 수 있는 기술에 대한 연구 필요성이 대두되고 있다. 본 연구에서는 염촉매 전기분해 공기살균기를 개발하여 공기살균기의 항균활성과 인간 세포주(HaCaT, BEAS-2B, THP-1)에 대한 세포독성을 통한 안전성을 조사하였다. 공기살균기에서 분무되는 증기를 각각 1, 3시간 동안 분무한 결과, Staphylococcus aureus는 각각 24.01, 57.34%, Bacillus subtilis는 10.89, 57.76%, Escherichia coli는 30.79, 36.59%, Pseudomonas aeruginosa는 34.80, 49.51%, Salmonella typhimurium는 39.40, 73.98%, 진균인 Candida albicans는 36.59, 53.05%의 높은 항균활성을 보여 염촉매 전기분해 공기살균기의 살균효능이 모든 종류의 시험균주에서 시간 의존적인 항균활성을 보였다. 공기살균기의 분무액을 0.1, 0.3, 1.0%의 농도로 처리하고 세포 생존율을 측정한 결과, 1.0%의 농도에서도 BEAS-2B 세포는 93.88%, HaCaT 세포는 97.01%, THP-1 세포는 86.56%로 관찰되어 인체 정상 세포주에서는 유의적인 세포독성을 나타내지 않았다. 또한 분무액을 1.0%의 농도로 처리하고 현미경으로 관찰한 세포주의 형태학적 변화, PCR을 이용한 세포사멸관련 유전자(Bcl-2, Bax)의 발현 분석, FACS를 이용한 세포 내 ROS의 생성 변화 등에서 분무액을 처리하지 않은 대조군과 비교하였을 때 유의적인 변화가 없었다. 따라서 시험균주에 대한 항균활성, 3가지의 인간 세포주에 대한 세포독성, 세포의 형태적 변화, 유독한 반응성 산소(ROS) 생성, 세포사멸관련 유전자의 발현 등에서 유의미한 변화가 확인되지 않아 공기살균기의 살균 효능과 사용상 안전성을 확인할 수 있었다. 최종적으로 다중이용시설에서의 공기살균기의 효능을 조사하기 위하여 밀폐된 실내에서 공기살균기를 20시간 동안 가동한 후, 공기 중의 낙하균을 포집하여 배양한 결과, 총세균은 89.4%, 대장균과 진균은 100.0% 제거되는 것을 확인하였다. 결론적으로 개발된 염촉매 전기분해 공기살균기는 인체에 대한 독성이 없으면서 실내 오염원인 미생물들을 효과적으로 제거할 수 있음을 확인하였다.

• Tuberculosis and Respiratory Diseases
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• 제82권2호
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• pp.133-142
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• 2019

Background: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. Methods: Mouse AE II cell line MLE-12 were exposed to $1-10{\mu}g/mL$ BLM and $0.01-100{\mu}M$ baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. Results: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor ${\alpha}$, and transforming growth factor ${\beta}1$. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. Conclusion: BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.6871-6876
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• 2015

Background: The meningeal hemangiopericytoma (MHPC) is a vascular tumor arising from pericytes. Most intracranial MHPCs resemble meningiomas (MNGs) in their clinical presentation and histological features and may therefore be misdiagnosed, despite important differences in prognosis. Materials and Methods: We report 8 cases of MHPC and 5 cases of MNG collected from 2007 to 2011 from the Neuro-Surgery and Histopathology departments. All 13 samples were re reviewed by two independent pathologists and investigated by immunohistochemistry (IHC) using mesenchymal, epithelial and neuro-glial markers. Additionally, we screened all tumors for a large panel of chromosomal alterations using multiplex ligation probe amplification (MLPA). Presence of the NAB2-STAT6 fusion gene was inferred by immunohistochemical staining for STAT6. Results: Compared with MNG, MHPCs showed strong VIM (100% of cases), CD99 (62%), bcl-2 (87%), and p16 (75%) staining but only focal positivity with EMA (33%) and NSE (37%). The p21 antibody was positive in 62% of MHPC and less than 1% in all MNGs. MLPA data did not distinguish HPC from MNG, with PTEN loss and ERBB2 gain found in both. By contrast, STAT6 nuclear staining was observed in 3 MHPC cases and was absent from MNG. Conclusions: MNG and MHPC comprise a spectrum of tumors that cannot be easily differentiated based on histopathology. The presence of STAT6 nuclear positivity may however be a useful diagnostic marker.

• IMMUNE NETWORK
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• 제5권1호
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• pp.23-29
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• 2005

Background: Interleukin-7 receptor (IL-7R) ${\alpha}$-deficient mice have small numbers of B cells and ${\alpha}{\beta}$T cells in periphery, they totally lack ${\gamma}{\delta}$T cells. In addition, the V-J recombination and transcription of TCR ${\gamma}$ genes is also severely impaired in IL-7R ${\alpha}$-deficient mice. Stat5, a signaling molecule of the IL-7R, induces germline transcription in the TCR ${\gamma}$ locus, and promotes V-J recombination and ${\gamma}{\delta}$T cell development. However, the roles for IL-7R signaling pathway in thymic or extrathymic ${\gamma}{\delta}$T cell development are largely unknown. Methods: To clarify the role of the IL-7 receptor in proliferation and survival of ${\gamma}{\delta}$T cells, we introduced the TCR ${\gamma}{\delta}$ transgene, $V_{{\gamma}2}/V{\delta}_5$, into IL-7R ${\alpha}$-deficient mice, and investigated the development of ${\gamma}{\delta}$T cells. Results: We found that $V_{{\gamma}2}/V{\delta}_5$ transgene restored ${\gamma}{\delta}$T cells in the epithelium of the small intestine (IEL) but not in the thymus and the spleen. Further addition of a bcl-2 transgene resulted in partial recovery of ${\gamma}{\delta}$T cells in the thymus and the spleen of these mice. Conclusion: Taken together, this study revealed that the IL-7R ${\alpha}$ is indispensable for proliferation and survival mainly in thymic ${\gamma}{\delta}$T cell development.

• The Korean Journal of Physiology and Pharmacology
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• 제21권6호
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• pp.625-632
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• 2017

Familial Parkinson's disease (PD) has been linked to point mutations and duplication of the ${\alpha}$-synuclein (${\alpha}$-syn) gene. Mutant ${\alpha}$-syn expression increases the vulnerability of neurons to exogenous insults. In this study, we developed a new PD model in the transgenic mice expressing mutant hemizygous (hemi) or homozygous (homo) A53T ${\alpha}$-synuclein (${\alpha}$-syn Tg) and their wildtype (WT) littermates by treatment with sub-toxic (10 mg/kg, i.p., daily for 5 days) or toxic (30 mg/kg, i.p., daily for 5 days) dose of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Tyrosine hydroxylase and Bcl-2 levels were reduced in the ${\alpha}$-syn Tg but not WT mice by sub-toxic MPTP injection. In the adhesive removal test, time to remove paper was significantly increased only in the homo ${\alpha}$-syn Tg mice. In the challenging beam test, the hemi and homo ${\alpha}$-syn Tg mice spent significantly longer time to traverse as compared to that of WT group. In order to find out responsible proteins related with vulnerability of mutant ${\alpha}$-syn expressed neurons, DJ-1 and ubiquitin enzyme expressions were examined. In the SN, DJ-1 and ubiquitin conjugating enzyme, UBE2N, levels were significantly decreased in the ${\alpha}$-syn Tg mice. Moreover, A53T ${\alpha}$-syn overexpression decreased DJ-1 expression in SH-SY5Y cells. These findings suggest that the vulnerability to oxidative injury such as MPTP of A53T ${\alpha}$-syn mice can be explained by downregulation of DJ-1.

• Journal of Microbiology and Biotechnology
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• 제28권6호
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• pp.1007-1021
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• 2018

Cancer represents one of the most significant threats to human health on a global scale. Hence, the development of effective cancer prevention strategies, as well as the discovery of novel therapeutic agents against cancer, is urgently required. In light of this challenge, this research aimed to evaluate the effects of several potent bioactive peptides and proteins contained in crocodile white blood cell extract (cWBC) against LU-1, LNCaP, PC-3, MCF-7, and CaCo-2 cancer cell lines. The results demonstrate that 25, 50, 100, and $200{\mu}g/ml$ cWBC exhibits a strong cytotoxic effect against all investigated cell lines ($IC_{50}$ $70.34-101.0{\mu}g/ml$), while showing no signs of cytotoxicity towards noncancerous Vero and HaCaT cells. Specifically, cWBC treatment caused a significant reduction in the cancerous cells' colony forming ability. A remarkable suppression of cancerous cell migration was observed after treatment with cWBC, indicating potent antimetastatic properties. The mechanism involved in the cancer cell cytotoxicity of cWBC may be related to apoptosis induction, as evidenced by typical apoptotic morphology features. Moreover, certain cWBC concentrations induced significant overproduction of ROS and significantly inhibited the $S-G_2/M$ transition in the cancer cell. The molecular mechanisms of cWBC in apoptosis induction were to decrease Bcl-2 and XIAP expression levels and increase the expression levels of caspase-3, caspase-8, and p53. These led to a decrease in the expression level of the cell cycle-associated gene cyclin-B1 and the arrest of cell population growth. Consequently, these findings demonstrate the prospect of the use of cWBC for cancer therapy.

• Animal Bioscience
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• 제35권9호
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• pp.1340-1350
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• 2022

Objective: Luxi Black Head sheep (LBH) is the first crossbreed specialized for meat production and was developed by crossbreeding Black Head Dorper sheep (DP) and Small Tailed Han sheep (STH) in the farming areas of northern China. Research on the genomic variations and selection signatures of LBH caused by continuous artificial selection is of great significance for identifying the genetic mechanisms of important traits of sheep and for the continuous breeding of LBH. Methods: We explored the genetic relationships of LBH, DP, and several Mongolian sheep breeds by constructing phylogenetic tree, principal component analysis and linkage disequilibrium analysis. In addition, we analysed 29 whole genomes of sheep. The genome-wide selection signatures have been scanned with four methods: heterozygosity (H_P), fixation index (F_ST), cross-population extended haplotype homozygosity (XP-EHH) and the nucleotide diversity (𝜃_π) ratio. Results: The genetic relationships analysis showed that LBH appeared to be an independent cluster closer to DP. The candidate signatures of positive selection in sheep genome revealed candidate genes for developmental process (HoxA gene cluster, BCL2L11, TSHR), immunity (CXCL6, CXCL1, SKAP2, PTK6, MST1R), growth (PDGFD, FGF18, SRF, SOCS2), and reproduction (BCAS3, TRIM24, ASTL, FNDC3A). Moreover, two signalling pathways closely related to reproduction, the thyroid hormone signalling pathway and the oxytocin signalling pathway, were detected. Conclusion: The selective sweep analysis of LBH genome revealed candidate genes and signalling pathways associated with developmental process, immunity, growth, and reproduction. Our findings provide a valuable resource for sheep breeding and insight into the mechanisms of artificial selection.

• 생명과학회지
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• 제20권7호
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• pp.1054-1065
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• 2010

생약복합물인 SH21B는 황금(Scutellaria baicalensis Georgi), 행인(Prunus armeniaca Maxim), 마황(Ephedra sinica Stapf), 석창포(Acorus gramineus Soland), 포황(Typha orientalis Presl), 원지(Polygala tenuifolia Willd), 하엽(Nelumbo nucifera Gaertner)의 혼합(비율 3:3:3:3:3:2:2)으로 이루어졌다. SH21B는 예로부터 한의학에서 비만의 치료에 사용되어 왔으나 자세한 분자적 메커니즘과 효능에 대한 연구는 이루어지지 않았다. 본 연구진은 선행연구를 통해 SH21B가 지방세포의 분화에서 adipogenesis (지방세포형성)와 관련된 유전자를 조절하여 중성지방의 축적을 억제함을 밝혔다. 본 연구에서는, microarray 기술을 이용하여 adipogenesis의 in vitro 모델인, 3T3-L1 세포에서 SH21B에 의한 지방세포형성 억제의 분자적 기작을 보다 상세하게 연구하고자 하였다. 전지방세포, 분화된 세포 그리고 SH21B에 의해 분화가 억제된 세포의 각각의 유전자 발현을 분석하기 위해 각 시료들에서 total RNA를 분리하여 cDNA를 합성한 후 microarray에 적용시켰다. 그 결과, 각각의 시료들의 비교에서 2배 이상의 유의한 발현 변화를 가지는 2,568개의 유전자를 확보하였다. 이 유전자들에 대해 Hierarchical clustering과 K-means clustering 분석을 진행하였고 서로 다른 양상을 가지는 9개의 군집(cluster)들을 분류하였다. 그 중, SH21B의 첨가에 의해 뚜렷하게 감소(cluster 4, cluster 6 및 cluster 9)하거나 반대로 뚜렷하게 증가(cluster 7와 cluster 8)하는 양상을 보이는 군집들을 따로 선별하여 그 군집들에 포함되어 있는 유전자들을 분석하였다. 선택 된 5개의 군집에는 지방세포형성과 세포증식에 관련된 유전자가 다수 포함되어 있었다. Cluster 4, cluster 6 그리고 cluster 9에는 peroxisome proliferator activated receptor gamma $\gamma$ ($PPAR{\gamma}$), CCAAT/enhancer binding protein $\alpha$ (C/$EBP{\alpha}$), sterol regulatory element binding transcription factor 1 (SREBF1), adiponectin (ADIPOQ), fatty acid synthase (FASN), lipoprotein lipase (LPL) 등의 지방세포형성 유도 및 관련 인자와 B-cell leukemia/lymphoma6 (BCL6), retinoblastoma 1 (RB1), cyclin-dependent kinase inhibitor 2C (CDKN2c), ras homolog gene family, member B (RHOB) 등의 많은 세포증식 억제 유전자가 포함되었다. 이와는 반대로, cluster 7과 cluster 8에는 $\beta$-catenin, cyclin D1 (CCND1), WNT1 inducible signaling pathway protein 2 (WISP2) 등과 같은 지방 세포형성 억제 조절자와 MARCKS-like1 (MARCKSL1), colony stimulating factor 1 (CSF1), discoidin domain receptor family, member 2 (DDR2), leukemia inhibitory factor receptor (LIFR) 등의 세포증식을 유도하는 조절자가 다수 포함되었다. 결론적으로, 이러한 결과들은 SH21B가 지방세포형성과 관련된 조절자 및 세포증식과 관련 된 조절자들의 유전자 발현을 조절하여 지방세포형성을 억제함을 제시한다.