• Journal of Veterinary Science
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• 제21권3호
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• pp.50.1-50.16
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• 2020

Background: Porcine parvovirus (PPV) is a single-stranded DNA virus that causes porcine reproductive failure. It is of critical importance to study PPV pathogenesis for the prevention and control of the disease. NS1, a PPV non-structural protein, is participated in viral DNA replication, transcriptional regulation, and cytotoxicity. Our previous research showed that PPV can activate nuclear factor kappa B (NF-κB) signaling pathway and then up-regulate the expression of interleukin (IL)-6. Objectives: Herein, the purpose of this study is to determine whether the non-structural protein NS1 of PPV also has the same function. Methods: Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay, western blot, immunofluorescence assay and small interfering RNA (siRNA) were used. Results: Our findings demonstrated that PPV NS1 protein can up-regulate the expression levels of IL-6 and tumor necrosis factor-alpha in a dose-dependent manner. Moreover, PPV NS1 protein was found to induce the phosphorylation of IκBα, then leading to the phosphorylation and nuclear translocation of NF-κB. In addition, the NS1 protein activated the upstream pathways of NF-κB. Meanwhile, TLR2-siRNA assay showed TLR2 plays an important role in the activation of NF-κB signaling pathway induced by PPV-NS1. Conclusions: These findings indicated that PPV NS1 protein induced the up-regulated of IL-6 expression through activating the TLR2 and NF-κB signaling pathways. In conclusion, these findings provide a new avenue to study the innate immune mechanism of PPV infection.

• Journal of Animal Science and Technology
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• 제66권1호
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• pp.204-218
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• 2024

Elsholtzia fruticosa (EF) is present in tropical regions throughout South Asian countries as well as the Himalayas. Although it has been used as a traditional medicine to treat digestive, respiratory, and inflammatory issues, its effect on preadipocyte differentiation is unknown. In this study, we examined the effects of a methanol extract prepared from EF on the differentiation of 3T3-L1 preadipocytes. Cell differentiation was assessed by microscopic observation and oil-red O staining. The expression of adipogenic and lipogenic genes, including PPARγ and C/EBPα, was measured by western blot analysis and quantitative real-time polymerase chain reaction (qRT-PCR), to provide insight into adipogenesis and lipogenesis mechanisms. The results indicated that EF promotes the differentiation of 3T3-L1 preadipocytes, with elevated lipid accumulation occurring in a concentration-dependent manner without apparent cytotoxicity. EF enhances the expression of adipogenic and lipogenic genes, including PPARγ, FABP4, adiponectin, and FAS, at the mRNA and protein levels. The effect of EF was more pronounced during the early and middle stages of 3T3-L1 cell differentiation. Treatment with EF decreased C/EBP homologous protein (CHOP) mRNA and protein levels, while increasing C/EBPα and PPARγ expression. Treatment with EF resulted in the upregulation of cyclin E and CDK2 gene expression within 24 h, followed by a decrease at 48 h, demonstrating the early-stage impact of EF. A concomitant increase in cyclin-D1 levels was observed compared with untreated cells, indicating that EF modulates lipogenic and adipogenic genes through intricate mechanisms involving CHOP and cell cycle pathways. In summary, EF induces the differentiation of 3T3-L1 preadipocytes by increasing the expression of adipogenic and lipogenic genes, possibly through CHOP and cell cycle-dependent mechanisms.

• Asian-Australasian Journal of Animal Sciences
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• 제32권8호
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• pp.1095-1103
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• 2019

Objective: Among stress responses, the unfolded protein response (UPR) is a well-known mechanism related to endoplasmic reticulum (ER) stress. ER stress is induced by a variety of external and environmental factors such as starvation, ischemia, hypoxia, oxidative stress, and heat stress. Inositol requiring enzyme $1{\alpha}$ ($IRE1{\alpha}$)-X-box protein 1 (XBP1) is the most conserved pathway involved in the UPR and is the main component that mediates $IRE1{\alpha}$ signalling to downstream ER-associated degradation (ERAD)- or UPR-related genes. XBP1 is a transcription factor synthesised via a novel mechanism called 'frame switch splicing', and this process has not yet been studied in the horse XBP1 gene. Therefore, the aim of this study was to confirm the frame switch splicing of horse XBP1 and characterise its dynamics using Thoroughbred muscle cells exposed to heat stress. Methods: Primary horse muscle cells were used to investigate heat stress-induced frame switch splicing of horse XBP1. Frame switch splicing was confirmed by sequencing analysis. XBP1 amino acid sequences and promoter sequences of various species were aligned to confirm the sequence homology and to find conserved cis-acting elements, respectively. The expression of the potential XBP1 downstream genes were analysed by quantitative real-time polymerase chain reaction. Results: We confirmed that splicing of horse XBP1 mRNA was affected by the duration of thermal stress. Twenty-six nucleotides in the mRNA of XBP1 were deleted after heat stress. The protein sequence and the cis-regulatory elements on the promoter of horse XBP1 are highly conserved among the mammals. Induction of putative downstream genes of horse XBP1 was dependent on the duration of heat stress. We confirmed that both the mechanisms of XBP1 frame switch splicing and various binding elements found in downstream gene promoters are highly evolutionarily conserved. Conclusion: The frame switch splicing of horse XBP1 and its dynamics were highly conserved among species. These results facilitate studies of ER-stress in horse.

• 미생물학회지
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• 제50권3호
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• pp.185-190
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• 2014

원유시료에서 분리한 대장균의 quinolone 항생제 내성비율과 그 내성 결정인자를 분석하였다. 원유시료에서 대장균을 분리하고 quinolone 항생제인 nalidixic acid와 ciprofloxacin에 대한 MIC값을 결정하였으며 내성균을 대상으로 염색체상에 있는 quinolone 내성 결정부위(quinolone resistant determining region, QRDR)인 gyrA, gyrB, parC, pareE의 염기서열 분석, 플라스미드상에 존재하는 내성유전자(plasmid mediated quinolone resistant, PMQR) qnrA, qnrB, qnrS, aac(6')-lb-cr, qepA의 분석 및 약물 유출펌프 유전자인 acrB의 발현을 비교 분석하였다. 그 결과 총 487개의 대장균군 세균중 9개의 균이 nalidixic acid에 내성임을 확인하였으며($MIC{\geq}64{\mu}g/ml$) 이중 6개 균주가 ciprofloxacin에도 내성임을 확인하였다(MIC $4-16{\mu}g/ml$)). 9개의 내성 균주 모두 QRDR의 gyrA 영역 codon 83에 변이(S83L)를 갖고 있었으며 그 중 2균주는 codon 83과 87 (S83L and D87N)에 이중 돌연변이를 갖고 있었다. 한편 9균주 중 3개의 균주에서 parC 영역 codon 80 (S80I)에 변이를 갖고 있었다. 플라스미드 상에 존재하는 내성유전자인 qnrA, qnrB, qnrS, aac(6')-lb-cr 및 qepA 유전자는 존재하지 않았으며 AcrAB-TolC efflux pump 유전자인 acrB 유전자가 대조균인 E. coli ATCC 25922와 비교하여 ciprofloxacin 내성 균주 6균주 중 4균주에서 유의적으로 과발현(2.15-5.74배) 되고 있음을 확인하였다.

• Genes and Genomics
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• 제40권11호
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• pp.1181-1197
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• 2018

Tropical plant rubber tree (Hevea brasiliensis) is the sole source of commercial natural rubber and low-temperature stress is the most important limiting factor for its cultivation. To characterize the gene expression profiles of H. brasiliensis under the cold stress and discover the key cold stress-induced genes. Three cDNA libraries, CT (control), LT2 (cold treatment at $4^{\circ}C$ for 2 h) and LT24 (cold treatment at $4^{\circ}C$ for 24 h) were constructed for RNA sequencing (RNA-Seq) and gene expression profiling. Quantitative real time PCR (qRT-PCR) was conducted to validate the RNA-Seq and gene differentially expression results. A total of 1457 and 2328 differentially expressed genes (DEGs) in LT2 and LT24 compared with CT were respectively detected. Most significantly enriched KEGG pathways included flavonoid biosynthesis, phenylpropanoid biosynthesis, plant hormone signal transduction, cutin, suberine and wax biosynthesis, Pentose and glucuronate interconversions, phenylalanine metabolism and starch and sucrose metabolism. A total of 239 transcription factors (TFs) were differentially expressed following 2 h or/and 24 h of cold treatment. Cold-response transcription factor families included ARR-B, B3, BES1, bHLH, C2H, CO-like, Dof, ERF, FAR1, G2-like, GRAS, GRF, HD-ZIP, HSF, LBD, MIKC-MADS, M-type MADS, MYB, MYB-related, NAC, RAV, SRS, TALE, TCP, Trihelix, WOX, WRKY, YABBY and ZF-HD. The genome-wide transcriptional response of rubber tree to the cold treatments were determined and a large number of DEGs were characterized including 239 transcription factors, providing important clues for further elucidation of the mechanisms of cold stress responses in rubber tree.

• 한국양봉학회지
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• 제32권3호
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• pp.155-162
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• 2017

로얄제리 생산성 우수계통 선발을 위해 국내 육성서양종꿀벌 원종 계통인 A와 C에 대한 로얄제리 생산성 및 접수율 평가를 수행하였다. 그 결과 꿀벌 계통C의 로얄제리 생산량은 $33.7{\pm}7.41g$로A 계통에 비해 높은 것으로 드러났으며, 접수율 또한 $87.8{\pm}7.5%$로 A 계통에 비해 높은 것으로 확인되었다. 이들 계통으로부터 생산된 로얄제리 trans-10-hydroxy-2-decenoic acid (10-HDA) 함량을 분석한 결과 꿀벌 계통 C는 4.28%(${\pm}0.3$)으로, A 계통에 비해 높은 것으로 나타났다. 일령에 따른 major royal jelly proteins(MRJPs) 전사체 발현량을 비교한 결과 10일령의 일벌의 경우 C 계통이 A 계통에 비해 MRJP 발현량이 큰 폭으로 증가했음을 확인 할 수 있었으며, 15일령의 일벌의 MRJP 발현량 또한 C계통이 A 계통에 비해 높은 것을 확인할 수 있었다. 이러한 결과를 통해 본 연구에서는 국내 육성 서양종꿀벌 기본종 계통 중 로얄제리 생산성이 가장 우수한 계통은 C 계통인 것으로 평가 되었으며, 향후 로얄제리 생산성 우수 계통 선발을 위한 기초 자료로 제공 될 수 있을 것이라 기대하고 있다.

• Animal Bioscience
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• 제35권10호
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• pp.1489-1498
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• 2022

Objective: The objective of this study was to identify polymorphism in olfactomedin like 3 (OLFML3) gene, and association analysis with meat quality, carcass characteristics, retail meat cut, and fatty acid composition in sheep, and expression quantification of OLFML3 gene in phenotypically divergent sheep. Methods: A total of 328 rams at the age of 10 to 12 months with an average body weight of 26.13 kg were used. A novel polymorphism was identified using high-throughput sequencing in sheep and genotyping of OLFML3 polymorphism was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Among 328 rams, 100 rams representing various sheep genotypes were used for association study and proc general linear model was used to analyse association between genotypes and phenotypic traits. Quantitative real-time polymerase chain reaction (qRT-PCR) was used for the expression analysis of OLFML3 mRNA in phenotypically divergent sheep population. Results: The findings revealed a novel polymorphism in the OLFML3 gene (g.90317673 C>T). The OLFML3 gene revealed three genotypes: CC, CT, and TT. The single nucleotide polymorphism (SNP) was found to be significantly (p<0.05) associated with meat quality traits such as tenderness and cooking loss; carcass characteristics such as carcass length; retail meat cut such as pelvic fat in leg, intramuscular fat in loin and tenderloin, muscle in flank and shank; fatty acids composition such as tridecanoic acid (C13:0), palmitoleic acid (C16:1), heptadecanoic acid (C17:0), ginkgolic acid (C17:1), linolenic acid (C18:3n3), arachidic acid (C20:0), eicosenoic acid (C20:1), arachidonic acid (C20:4n6), heneicosylic acid (C21:0), and nervonic acid (C24:1). The TT genotype was associated with higher level of meat quality, carcass characteristics, retail meat cut, and some fatty acids composition. However, the mRNA expression analysis was not different among genotypes. Conclusion: The OLFML3 gene could be a potential putative candidate for selecting higher quality sheep meat, carcass characteristics, retail meat cuts, and fatty acid composition in sheep.

• Journal of Nutrition and Health
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• 제56권5호
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• pp.455-468
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• 2023

본 연구에서는 잠재적인 기능성 소재로의 개발 가능성을 검증하기 위하여 미선나무 잎 추출물의 항산화능과 항염증 효능을 확인하였다. 시료의 총 폴리페놀함량, DPPH 및 ABTS 라디칼 소거능, FRAP value를 측정하여 미선나무 잎 추출물의 항산화 활성을 확인하였으며 또한 RAW264.7 세포에 미선나무 잎 추출물 처리 후 HO-1의 발현이 증가하는 것을 통해 미선나무 잎 추출물의 항산화 기전을 확인하였다. 한편, LPS로 유도된 염증 상태의 RAW264.7 세포에서 미선나무 잎 추출물은 NF-κB 활성 억제를 통한 NO 생성 억제, iNOS, COX-2 발현 억제 및 염증성 사이토카인의 발현과 생성을 억제하는 것을 확인하였다. 이러한 본 연구 결과는 향후 미선나무 잎 추출물의 기능성 소재로의 개발을 위한 기초자료 마련에 그 의의가 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8893-8900
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• 2014

Dysregulated expression of microRNAs (miRNAs) has been shown to be closely associated with tumor development, progression, and carcinogenesis. However, their clinical implications for gastric cancer remain elusive. To investigate the hypothesis that genome-wide alternations of miRNAs differentiate gastric cancer tissues from those matched adjacent non-tumor tissues (ANTTs), miRNA arrays were employed to examine miRNA expression profiles for the 5-pair discovery stage, and the quantitative real-time polymerase chain reaction (qRTPCR) was applied to validate candidate miRNAs for 48-pair validation stage. Furthermore, the relationship between altered miRNA and clinicopathological features and prognosis of gastric cancer was explored. Among a total of 1,146 miRNAs analyzed, 16 miRNAs were found to be significantly different expressed in tissues from gastric cancer compared to ANTTs (p<0.05). qRT-PCR further confirmed the variation in expression of miR-193b and miR-196a in the validation stage. Down-expression of miR-193b was significantly correlated with Lauren type, differentiation, UICC stage, invasion, and metastasis of gastric cancer (p<0.05), while over-expression of miR-196a was significantly associated with poor differentiation (p=0.022). Moreover, binary logistic regression analysis demonstrated that the UICC stage was a significant risk factor for down-expression of miR-193b (adjusted OR=8.69; 95%CI=1.06-56.91; p=0.043). Additionally, Kaplan-Meier survival curves indicated that patients with a high fold-change of down-regulated miR-193b had a significantly shorter survival time (n=19; median survival=29 months) compared to patients with a low fold-change of down-regulated miR-193b (n=29; median survival=54 months) (p=0.001). Overall survival time of patients with a low fold-change of up-regulated miR-196a (n=27; median survival=52 months) was significantly longer than that of patients with a high fold-change of up-regulated miR-196a (n=21; median survival=46 months) (p=0.003). Hence, miR-193b and miR-196a may be applied as novel and promising prognostic markers in gastric cancer.

• 한국해양생명과학회지
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• 제6권2호
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• pp.73-79
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• 2021

비스페놀A(BPA)는 대표적인 내분비계 교란물질로 광범위한 사용으로 인해 환경 내에서 지속적으로 검출됨에 따라 인간을 비롯한 다양한 생물에서 성장, 발생, 생식 등에 유해한 영향을 미치는 것으로 알려져 있다. 따라서 BPA를 대체하기 위한 구조 유사체들이 개발되어 널리 사용되고 있으나 이러한 대체제들이 내분비계 교란 작용을 갖는지에 대한 연구가 필요하다. 본 연구에서는 BPA와 그 구조 유사체인 BPS와 BPF에 노출시킨 기수산 물벼룩 Diaphanosoma celebensis에서 탈피과정에 관여하는 ecdysteroid 합성(nvd, cyp314a1), receptors (EcRA, EcRB, USP, ERR), 그리고 하위 경로에 있는 유전자(HR3, E75, Vtg, VtgR)의 시간 별 발현 변화를 조사하였다. nvd와 cyp314a1 유전자의 발현은 BPA 보다 BPF에서 6시간 일찍 발현이 증가하는 양상을 보인 반면, BPS의 경우에는 이들 유전자의 발현이 24시간 내내 감소하는 양상을 보였다. BPA와 BPF 노출 시 EcR 유전자들의 발현 양상도 이와 유사한 경향을 보였다. ERR 유전자의 발현은 BPF와 BPS에서 BPA 보다 6시간 일찍 발현이 증가하는 양상을 보였고, HR3, E75, VtgR의 유전자 발현도 노출군에서 시간 차이는 있지만 유의하게 증가하는 양상을 보였다. 반면 Vtg는 24시간 이내에서는 크게 증가하지는 않았다. 이러한 결과는 BPA 뿐 아니라 BPF와 BPS도 탈피에 관여하는 호르몬의 합성 및 조절 경로의 유전자의 발현을 조절할 수 있으며, 서로 다른 기전으로 기수산 물벼룩의 내분비계를 교란시킬 수 있는 능력을 갖는다고 볼 수 있다. 본 연구는 비스페놀 구조 유사체가 기수산 물벼룩의 탈피과정에 관여하는 분자 경로 어떻게 영향을 미치는지를 이해하는데 도움이 될 것이다.