• 한국가금학회지
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• 제46권3호
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• pp.185-191
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• 2019

Nucleoporin 210(Nup210)는 근육 및 신경세포의 분화, 자기 면역 질환, 말초 T세포 항상성 등 여러 생리작용에 관여한다. 닭의 Nup210 유전자는 닭 신장조직에서 칼슘 의존성 차별 발현 유전자로 발굴되었으며, 닭의 대사 이상 질환과 Nup210의 관련 연구를 위해 Nup210 유전자의 분자유전학적 특성을 구명하고, 톨-유사수용체 3(Toll-like receptor 3(TLR3)) 자극에 의한 전사 조절을 연구하였다. 닭의 여러 조직과 배아 섬유아세포주인 DF-1 세포에서 Nup210 유전자의 전사 수준을 조사한 결과, 폐와 비장 조직에서 가장 높게 발현되었으며, Nup210의 발현은 TLR3 신호자극에 의해 증가함을 확인하였다. 또한 닭 Nup210 유전자가 코딩하는 단백질의 구조는 조류, 어류, 포유류를 포함한 여러 종과 매우 보존적이나 진화적으로 다른 포유류보다는 오리와 가장 가깝다고 추정되었다. 본 연구의 결과를 통해 닭 Nup210이 TLR3 신호시스템에 관여함을 확인하였고, 추가연구를 통해 바이러스 침입에 대한 닭 면역 메커니즘을 구명할 필요가 있다고 사료된다.

• 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.

• 현장농수산연구지
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• 제22권1호
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• pp.5-13
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• 2020

생체 조직 내에서 표지인자의 발견은 해당 세포의 특성과 기능을 이해하는 데 매우 중요하다. 기존에 밝혀진 돼지 정원세포의 표지인자로는 PGP9.5, PLZF, NANOG, SSEA1 등의 단백질이 알려져 있다. 본 연구에서는 최근 새로이 발굴된 돼지 정원세포 표지인자인 IGFBPs 의 기능을 분석하기 위해 IGFBPs 의 발현과 이를 조절하는 단백질인 PAPP-A 단백질의 발현을 5일령 돼지 정소에서 확인하였다. IGFBP 2, 3, 4, 6 의 발현이 돼지 정원세포 특이적으로 높게 나타났으며 PAPP-A의 발현은 세르톨리세포 특이적으로 나타났다. PAPP-A 의 발현을 PGP9.5, GATA4 등의 표지 인자와 함께 확인해 본 결과 PGP9.5를 발현하는 정원세포에서는 발현하지 않았으며, 세정관 내의 세르톨리세포 특이적으로 발현하였다. 이러한 사실로 미루어 볼 때 세르톨리세포에서 발현하는 PAPP-A 단백질은 정원세포에서 발현하는 IGFBPs 의 조절을 통하여 알려진 바와 같이 IGF axis 를 통해 정소내 세포들의 발달 및 분화를 조절할 것으로 판단된다.

• 대한본초학회지
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• 제29권4호
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• pp.53-59
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• 2014

Objectives : The purpose of this study was verification of the anti-inflammation and anti-oxidant effect of Cheongungdajosan-gamibang extract (CG) in mouse macrophage, RAW 264.7 cells. Methods : We have basically using LPS-stimulated RAW 264.7 cells. The cell toxicity was determined by MTT assay. To evaluate the anti-inflammatory effect of Cheongungdajosan-gamibang, amount of nitric oxide(NO) was measured using the NO detection kit and the IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). Also, free radical scavenging assay has tested for DPPH and ABTS radical activity as well as the contents of total polyphenol. Results : In this study, 96.6% or higher cell viability was observed in all tested groups from 1, 10, $100{\mu}/m{\ell}$ in RAW 264.7 cells. The RAW 264.7 cells were induced by lipopolysaccharide (LPS) and CG 1, 10, $100{\mu}/m{\ell}$. The CG decreased nitric oxide (NO) production activity dose dependently, especially at $100{\mu}/m{\ell}$ of 55%. The production of IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ were decreased by 51%, 78% and 35% in CG treated $100{\mu}g/m{\ell}$. CG showed dose-dependent suppression activity of reactive oxygen species (ROS) production, especially at $100{\mu}g/m{\ell}$ of 37%. DPPH radical scavenging activity and ABTS cation decolorization were activated over 86% and 88% in CG at $1,000{\mu}g/m{\ell}$ concentration. Conclusions : According to the results, we thought that CG showed anti-inflammatory and antioxidant activities on the RAW 264.7 cells in mouse macrophage. Therefore, this research is expected to provide the fundamental data about the natural material analysis of relating to the anti-inflammation and antioxidant.

• 대한본초학회지
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• 제28권5호
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• pp.7-11
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• 2013

Objectives : Seungmagalgeun-tang (SMGGT) is traditional medicine widely used for inflammatory disease and flu. But SMGGT exhibits potent anti-inflammatory activity with an unknown mechanism. To elucidate the molecular mechanisms of SMGGT water extract on pharmacological and biochemical actions in inflammation, we examined the effect of SMGGT on pro-inflammatory mediators in Phorbol-12-myristate-13-acetate (PMA)+A23187-stimulated mast cells. Methods : In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to measure the activation of MAPKs. Cells were treated with SMGGT 1 h prior to the addition of 50 nM of PMA and $1{\mu}M$ of A23187. Cell viability was measured by MTS assay. The investigation focused on whether SMGGT inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8) and mitogen-activated protein kinases (MAPKs) in PMA+A23187-stimulated mast cells. Results : SMGGT has no cytotoxicity at examined concentration (100, 250, and $500{\mu}g/ml$). Also, gene expression of IL-6 and IL-8 in HMC-1 cells stimulated by PMA+A23187 was down regulated by SMGGT. Furthermore, SMGGT suppressed the PMA+A23187-induced phosphorylation of extracellular signal-regulated kinase (ERK) and c-jun N-terminal Kinase(JNK). But, SMGGT could not regulate phosphorylation of p38 MAPK. Conclusions : These results suggest that SMGGT has inhibitory effects on PMA+A23187-induced IL-6 and IL-8 production. These inhibitory effects occur through blockades on the phosphorylation of ERK and JNK.

• 동의생리병리학회지
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• 제36권5호
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• pp.181-186
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• 2022

Atractylodes macrocephala (AM) and Amomum villosum (AV) are the most common herbs in Korean Medicine to treat digestive diseases. In this study, we investigated the cholesterol lowering effects of mixtures of AM and AV extracts on high cholesterol diet (HCD) induced dyslipidemia mouse model. We classified animals into six different groups; Group 1: Normal diet, Group 2: HCD, Group 3: AV extracts : AM extracts (1:1) (200 mg/kg) + HCD, Group 4: AV extracts : AM extracts (1:2) (200 mg/kg) + HCD, Group 5: AV extracts : AM extracts (1:3) (200 mg/kg) + HCD, Group 6: Simvastatin 40 mg/kg + HCD. After 4 weeks of oral administration of respective drugs, we checked body, liver and epididymal fatweights along with liver and serum triacylglyceride (TG) concentration, total and low density lipoprotein (LDL) cholesterol in serum. Moreover, 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase (HMGCR), LDL receptor (LDLR), and sterol regulatory element-binding protein 2 (SREBP2) were detected by RT PCR or western blot analysis. The overall results showed that mixtures of AM and AV extracts inhibited HCD-induced increases of total cholesterol and LDL cholesterol in serum. Those effects seem to be caused by AM and AV extracts through inhibition of HMGCR expression. And thus blood cholesterol is induced into the liver by increasing LDLR expression, which is regulated by SREBP2 transcrption factor. The cholesterol lowering effects and mechanism of mixtures of AM and AV extracts was similar to the statin. We have identified the potential mixtures of AM and AV extracts as a new treatment for dyslipidemia.

• 환경생물
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• 제41권1호
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• pp.1-13
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• 2023

잠두위조바이러스2 (BBWV2)는 경제적으로 중요한 원예, 관상작물에 심각한 피해를 주는 바이러스 중의 하나다. BBWV2의 넓은 기주 범위, 매개충 방제의 어려움 등 효과적인 치료제가 없기 때문에, 병을 예방하거나 저항성 품종의 개발 등을 위해서는 BBWV2의 새로운 분리주들의 특성 조사와 연구가 필요하다. 본 연구에서는 2019년 금산지역의 잎들깨 재배농가에서 분리된 BBWV2 분리주(BBWV2-GS-PF)의 유전학적·생물학적 특성을 구명하고 기존에 보고된 분리주들과의 특성을 비교하였다. 순수 분리된 BBWV2-GS-PF는 기존 분리주와 달리 들깨에서 심한 모자이크 증상과 고추에서 원형 반점 증상을 보였다. BBWV2-GS-PF의 유전자 계통분석 결과, 기존에 알려진 2개의 그룹과 약 76~80%의 비교적 낮은 유전자 상동성을 보이며 계통이 분화된 것을 확인할 수 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2021년도 춘계학술대회
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• pp.46-46
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• 2021

The first purpose of this study was to evaluate the scavenging capacity (SC) of DPPH and ABTS free radicals for ethanol extract (STR-E) and its active fractions from Sophora tonkinensis root (STR). Four different fractions from STR-E were prepared by using different types of solvents such as chloroform (STR-E-C), ethyl acetate (STR-E-EA), n-butanol (STR-E-B), and water (STR-E-W). STR-E-C showed the highest value of total phenolic content, while STR-E showed the highest value of total flavonoid and terpenoid content. In STR-E and its four fractions, STR-E-EA showed the strongest SC with the lowest SC50 values of the DPPH radicals and ABTS radicals. The second purpose of this study was to evaluate anti-inflammatory activity in the lipopolysaccharide (LPS)-induced RAW 264.7 macrophages treated with STR-E, STR-E-C, and STR-E-EA, respectively. No cytotoxic effect to RAW 264.7 cells was observed at 20 ~ 25 ㎍/ml of STR-E, 10 ㎍/ml of STR-E-C, and 5 ㎍/ml of the STR-E-EA, presenting cell viability values close to that of the untreated control (100%). STR-E, STR-E-C, and STR-E-EA significantly suppressed the LPS-induced nitric oxide (NO) in a dose-dependent manner. Results of reverse-transcription (RT)-qPCR analysis showed that the peak mRNA levels of IL-1β, TNF-α, iNOS, IL-6, and IL-10 were observed in the LPS-stimulated macrophages at 4 h, 2 h, 12 h, 12 h, and 12 h, respectively. The peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 were significantly reduced in the LPS-stimulated macrophages co-treated with 20 ㎍/ml and 25 ㎍/ml of STR-E, respectively. In the case of IL-10, its peak mRNA level slightly increased without statistical significance. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 10 ㎍/ml and 20 ㎍/ml of STR-E-C, respectively. In contrast, the peak mRNA level of IL-10 significantly increased at 8 h. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 5 ㎍/ml and 10 ㎍/ml of STR-E-EA, respectively. In contrast, the peak mRNA level of IL-10 increased at 4 h. Taken together, our data indicated that STR-E, STR-E-C, and STR-E-EA activate macrophages to secrete both pro-inflammatory and anti-inflammatory cytokines.

• Journal of Ginseng Research
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• 제48권3호
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• pp.298-309
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• 2024

Background: 20(S)-ginsenoside Rh2(GRh2), an effective natural histone deacetylase inhibitor, can inhibit acute myeloid leukemia (AML) cell proliferation. Lactate regulated histone lactylation, which has different temporal dynamics from acetylation. However, whether the high level of lactylation modification that we first detected in acute promyelocytic leukemia (APL) is associated with all-trans retinoic acid (ATRA) resistance has not been reported. Furthermore, Whether GRh2 can regulate lactylation modification in ATRA-resistant APL remains unknown. Methods: Lactylation and METTL3 expression levels in ATRA-sensitive and ATRA-resistant APL cells were detected by Western blot analysis, qRT-PCR and CO-IP. Flow cytometry (FCM) and APL xenograft mouse models were used to determine the effect of METTL3 and GRh2 on ATRA-resistance. Results: Histone lactylation and METTL3 expression levels were considerably upregulated in ATRA-resistant APL cells. METTL3 was regulated by histone lactylation and direct lactylation modification. Overexpression of METTL3 promoted ATRA-resistance. GRh2 ameliorated ATRA-resistance by downregulated lactylation level and directly inhibiting METTL3. Conclusions: This study suggests that lactylation-modified METTL3 could provide a promising strategy for ameliorating ATRA-resistance in APL, and GRh2 could act as a potential lactylation-modified METTL3 inhibitor to ameliorate ATRA-resistance in APL.

• Biomolecules & Therapeutics
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• 제32권1호
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• pp.154-161
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• 2024

Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder that causes progressive paralysis. L-Citrulline is a nonessential neutral amino acid produced by L-arginine via nitric oxide synthase (NOS). According to previous studies, the pathogenesis of ALS entails glutamate toxicity, oxidative stress, protein misfolding, and neurofilament disruption. In addition, L-citrulline prevents neuronal cell death in brain ischemia; therefore, we investigated the change in the transport of L-citrulline under various pathological conditions in a cell line model of ALS. We examined the uptake of [¹⁴C]L-citrulline in wild-type (hSOD1wt/WT) and mutant NSC-34/ SOD1^G93A (MT) cell lines. The cell viability was determined via MTT assay. A transport study was performed to determine the uptake of [¹⁴C]L-citrulline. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to determine the expression levels of rat large neutral amino acid transported 1 (rLAT1) in ALS cell lines. Nitric oxide (NO) assay was performed using Griess reagent. L-Citrulline had a restorative effect on glutamate induced cell death, and increased [¹⁴C]L-citrulline uptake and mRNA levels of the large neutral amino acid transporter (LAT1) in the glutamate-treated ALS disease model (MT). NO levels increased significantly when MT cells were pretreated with glutamate for 24 h and restored by co-treatment with L-citrulline. Co-treatment of MT cells with L-arginine, an NO donor, increased NO levels. NSC-34 cells exposed to high glucose conditions showed a significant increase in [¹⁴C]L-citrulline uptake and LAT1 mRNA expression levels, which were restored to normal levels upon co-treatment with unlabeled L-citrulline. In contrast, exposure of the MT cell line to tumor necrosis factor alpha, lipopolysaccharides, and hypertonic condition decreased the uptake significantly which was restored to the normal level by co-treating with unlabeled L-citrulline. L-Citrulline can restore NO levels and cellular uptake in ALS-affected cells with glutamate cytotoxicity, pro-inflammatory cytokines, or other pathological states, suggesting that L-citrulline supplementation in ALS may play a key role in providing neuroprotection.