• Asian-Australasian Journal of Animal Sciences
• /
• 제20권5호
• /
• pp.627-632
• /
• 2007

FoxO1, FoxO3a and FoxO4 belong to the FoxO gene family, which play important roles in the PI3K/PKB pathway. In this study, we cloned the porcine FoxO1, FoxO3a and FoxO4 sequences and assigned them to SSC11p11-15, SSC1p13 and SSC xq13 using somatic cell hybrid panel (SCHP) and radiation hybrid panel (IMpRH). RT-PCR results showed that these three genes are expressed in multiple tissues. Sequencing of PCR products from different breeds identified a synonymous T/C polymorphism in exon 2 of FoxO3a. This FoxO3a single nucleotide polymorphism (SNP) can be detected by AvaII restriction enzyme. The allele frequencies of this SNP were investigated in Dahuabai, Meishan, Tongcheng, Yushan, Large White, and Duroc pigs. Association of the genotypes with growth and carcass traits showed that different genotypes of FoxO3a were associated with carcass length and backfat thickness between 6th and 7th ribs (BTR) and drip loss (p<0.05).

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권4호
• /
• pp.499-509
• /
• 2008

Foxo1 plays an important role in the integration of hormone-activated signaling pathways with the complex transcriptional cascade that promotes preadipocyte differentiation of clonal cell lines from rodents. We isolated the full-length cDNA of porcine FoxO1 gene using RACE, confirmed by visual Northern blotting. The deduced amino acids indicated 94% and 90% identities with the corresponding human and mice aa. Analysis of the aa sequence, showed that it included a Forkhead domain (aa 167-247), a transmembrane structure domain (aa 90-113), a LXXLL motif (aa 469-473), and 51 Ser, 8 Thr, and 4 Tyr phosphorylation sites, indicating a potential important role for FoxO1 transcriptional activity in vivo. Using the IMpRH panel, we mapped FoxO1 gene to chromosome 11p13. Our data provide basic molecular information useful for the further investigation on the function of FoxO1 gene. Time-course analysis of FoxO1 expressions indicated that levels of mRNA and protein gradually increased from day 0 to 3, and it reached almost maximal level at day 3, then decreased from day 5 to 7 in porcine primary preadipocyte differentiation. After induction by IGF-1, GPDH activity and accumulation of lipid increased, however, expressions of FoxO1 mRNA and protein were inhibited in a dose dependent manner. These results suggest that FoxO1 takes part in porcine preadipocyte differentiation and expressions of FoxO1 were regulated by IGF-1.

• Journal of Microbiology and Biotechnology
• /
• 제28권4호
• /
• pp.520-526
• /
• 2018

Conessine, a steroidal alkaloid, is a potent histamine H3 antagonist with antimalarial activity. We recently reported that conessine treatment interferes with $H_2O_2$-induced cell death by regulating autophagy. However, the cellular signaling pathways involved in conessine treatment are not fully understood. Here, we report that conessine reduces muscle atrophy by interfering with the expression of atrophy-related ubiquitin ligases MuRF-1 and atrogin-1. Promoter reporter assay revealed that conessine treatment inhibits FoxO3a-dependent transcription, $NF-{\kappa}B$-dependent transcription, and p53-dependent transcription. We also showed by quantitative RT-PCR and western blot assays that conessine treatment reduced dexamethasone-induced expression of MuRF1 and atrogin-1. Finally, we demonstrated that conessine treatment reduced dexamethasone-induced muscle atrophy using differentiated C2C12 cells. These results collectively suggest that conessine is potentially useful in the treatment of muscle atrophy.

• 생명과학회지
• /
• 제26권4호
• /
• pp.439-445
• /
• 2016

콩잎은 골다공증 및 유방암 발생을 예방한다고 널리 보고되고 있다. 이를 바탕으로 콩잎낙엽 에탄올 추출물(SBFL)을 제조하여 암 전이와 관련 있는 세포침윤에 대한 효과를 조사하기 위하여 섬유아육종세포(HT1080)에서 SBFL이 항산화와 matrix metalloproteinases (MMPs)에 미치는 영향을 분석하였다. 본 연구에서 활성산소의 소거 효과에 대한 SBFL효과는 DPPH radical, 환원력 및 지질과산화실험으로 평가되었다, 본 연구에서 SBFL은 양성 대조군으로 사용된 vitamin C 및 vitamin E와 비교 시 우수한 항산화 효과를 보여주었다. 다음으로 SBFL의 세포 독성을 측정하기 위하여 MTT assay를 수행한 결과 16 µg/ml 이상의 농도에서 세포독성을 보여주었다. SBFL은 gelatin zymography 실험에서 phorbol 12-myristate 13-acetae (PMA) 혹은 phenazine methosulfate (PMS)로 자극된 암 전이에서 중요한 MMP-9의 활성을 감소시켰다. 특히 SBFL은 단백질 발현 실험에서 SOD-1, p-FoxO-1의 발현을 증가시켰다. 더욱이 vascular endothelial growth Factor (VEGF)로 자극된 세포 침윤이 SBFL처리에 의하여 억제되는 것으로 나타났다. 이러한 연구결과를 바탕으로 SBFL은 뛰어난 항산화 효과는 산화적 스트레스를 감소시키고 MMP-9의 활성과 세포침윤을 억제시켜 암 전이의 예방을 위한 유효성분으로 이용될 수 있다는 것을 암시하고 있다.

• 생명과학회지
• /
• 제30권2호
• /
• pp.211-220
• /
• 2020

CAR의 활성은 리간드 결합 뿐만 아니라, 세포외신호전달 경로를 통한 관련 조절인자들의 인산화, 전사 조절인자들과의 상호작용, 그리고 coactivators 및 corepressors의 동원, 분해 및 발현 등에 의해 조절되며, 이러한 CAR의 활성 조절은 또한 외인성 화학물질과 에너지 대사, 세포의 증식 및 사멸을 포함한 다양한 생리적 항상성 조절에 영향을 미친다. CAR는 ERK1/2의 신호전달경로에 의해 인산화되어 Hsp-90/CCRP와 복합체를 형성하여 세포질 내에 잔류하는 반면, PB는 ERK1/2를 억제하여 downstream 신호전달 조절인자들의 탈인산화를 유발하고, 활성화된 RACK-1/PP2A를 동원하여 CAR를 탈인산화 함으로써 핵 이동 및 전사 활성을 유도한다. CAR의 활성은 FoxO1 및 PGC-1α와의 cross-talk을 통하여 각각 전사 활성 억제와 ubiquitination을 통한 단백질 분해를 유도하여 당합성과정에 관여하는 PEPCK 및 G6Pase 유전자의 발현을 억제한다. CAR에 의한 지방의 합성과 산화 조절은 각각 PPARγ 및 PPARα와의 cross-talk에 의한 PGC-1α의 분해와 CPT-1의 발현 억제 또는 PGC-1α와의 결합을 통해 지방 합성 유전자의 발현 억제와 조직 특이적 산화 억제 또는 촉진으로 이루어진다. CAR는 FoxO1의 억제를 통한 p21의 발현 억제와 cyclin D1의 발현을 유도하여 세포 증식을 촉진하는 반면, GADD45B의 발현을 통한 MKK7과 JNK1의 활성을 억제하여 세포 사멸을 억제한다. 결론적으로, CAR는 세포외신호전달 경로와 세포내 조절인자들과의 다양한 상호작용을 통하여 외인성 화학물질의 대사뿐만 아니라 에너지 대사 및 세포의 성장과 사멸의 조절을 통한 항상성 유지에 관여한다.

• 한국축산식품학회지
• /
• 제25권1호
• /
• pp.71-77
• /
• 2005

본 연구는 재래종 세절 돼지고기의 저장 중 품질 특성을 구명하기 위하여 개량종과 비교하여 실시하였다. 시험용 돼지고기로 240일간 사육된 평균체중 75 kg의 재래돼지와 210일간 사육된 평균체중 105 kg의 개량돼지를 이용하였다. 도축 후 세절돈육을 4℃에서 9일간 저장하며 실시하였다. 일반성분의 조성을 보면 재래종 돼지고기가 개량종에 비해 조지방과 조단백질 함량이 높고, 수분과 조회분 함량은 낮았다. 재래종 돼지고기의 pH가 개량종에 비해 저장 5일부터는 낮았고, 보수력은 저장기간 내내 유의적으로 높았다(p<0.05). 재래종 돼지고기는 명도가 낮고 적색도가 높아 개량종에 비해 검붉은 색깔을 지니고 있다. 저장기간이 늘어날수록 CIE L/sup */, a/sup */, b/sup */와 h/sup O/값들은 감소하는 경향을 보였다. TBARS, POV, FOX값 모두 저장기간이 증가할수록 유의적으로 증가하였으며 특히 재래종이 저장기간이 길어질수록 현저한 증가현상을 나타내었다. 재래종 돼지고기의 지방산 조성비율을 보면 총 포화지방산과 stearic acid 함량이 개량종에 비해 유의적으로 높았다(p<0.05).

• Biomolecules & Therapeutics
• /
• 제29권3호
• /
• pp.311-320
• /
• 2021

Accumulation of reactive oxygen species (ROS) is associated with the development of various diseases. However, the molecular mechanisms underlying oxidative stress that lead to such diseases like autosomal dominant polycystic kidney disease (ADPKD) remain unclear. Here, we observed that oxidative stress markers were increased in Pkd1f/f:HoxB7-Cre mice. Forkhead transcription factors of the O class (FOXOs) are known key regulators of the oxidative stress response, which have been observed with the expression of FoxO3a in an ADPKD mouse model in the present study. An integrated analysis of two datasets for differentially expressed miRNA, such as miRNA sequencing analysis of Pkd1 conditional knockout mice and microarray analysis of samples from ADPKD patients, showed that miR-132-3p was a key regulator of FOXO3a in ADPKD. miR-132-3p was significantly upregulated in ADPKD which directly targeted FOXO3 in both mouse and human cell lines. Interestingly, the mitochondrial gene Gatm was downregulated in ADPKD which led to a decreased inhibition of Foxo3. Overexpression of miR-132-3p coupled with knockdown of Foxo3 and Gatm increased ROS and accelerated cyst formation in 3D culture. This study reveals a novel mechanism involving miR-132-3p, Foxo3, and Gatm that is associated with the oxidative stress that occurs during cystogenesis in ADPKD.

• Molecules and Cells
• /
• 제45권3호
• /
• pp.112-121
• /
• 2022

Calorie restriction (CR) and the activation of autophagy extend healthspan by delaying the onset of age-associated diseases in most living organisms. Because protein kinase CK2 (CK2) downregulation induces cellular senescence and nematode aging, we investigated CK2's role in CR and autophagy. This study indicated that CR upregulated CK2's expression, thereby causing SIRT1 and AMP-activated protein kinase (AMPK) activation. CK2α overexpression, including antisense inhibitors of miR-186, miR-216b, miR-337-3p, and miR-760, stimulated autophagy initiation and nucleation markers (increase in ATG5, ATG7, LC3BII, beclin-1, and Ulk1, and decrease in SQSTM1/p62). The SIRT1 deacetylase, AKT, mammalian target of rapamycin (mTOR), AMPK, and forkhead homeobox type O (FoxO) 3a were involved in CK2-mediated autophagy. The treatment with the AKT inhibitor triciribine, the AMPK activator AICAR, or the SIRT1 activator resveratrol rescued a reduction in the expression of lgg-1 (the Caenorhabditis elegans ortholog of LC3B), bec1 (the C. elegans ortholog of beclin-1), and unc-51 (the C. elegans ortholog of Ulk1), mediated by kin-10 (the C. elegans ortholog of CK2β) knockdown in nematodes. Thus, this study indicated that CK2 acted as a positive regulator in CR and autophagy, thereby suggesting that these four miRs' antisense inhibitors can be used as CR mimetics or autophagy inducers.

• IMMUNE NETWORK
• /
• 제20권6호
• /
• pp.50.1-50.11
• /
• 2020

Osteoporosis is prevalent in elderly women and it may cause dental implant failure. In particular, estrogen deficiency in postmenopausal women leads to higher rates of osteoporosis prevalence. Immune cell-mediated effects involving the development of osteoporosis have been studied previously; however, the role of IL-10-producing regulatory B (B10) cells in osteoporosis is largely unclear. Here, we examined the role of B10 cells in osteoporosis. C57BL/6 mice were subjected to ovariectomy (OVX). Fifteen weeks after OVX surgery, the first molar of the right maxillary was extracted, and twenty-four weeks after OVX surgery, serous progression of osteoporosis was observed in the alveolar bone. Moreover, the proportion of CD19⁺CD5⁺CD1d^high regulatory B cells, B10, and CD4⁺CD25⁺FoxP3⁺ regulatory T cells from the spleen of OVX mice decreased during the progression of osteoporosis, compared to controls. In contrast to regulatory cells, IL-17-producing Th (Th17) cell levels were increased in OVX mice. Adoptive transfer of B10 cells to OVX mice led to a decrease in Th17 cell abundance and inhibited the development of osteoporosis in the alveolar bone from OVX mice. Thus, our results suggest that B10 cells may help suppress osteoporosis development.

• Journal of Ginseng Research
• /
• 제47권6호
• /
• pp.726-734
• /
• 2023

Background: Skeletal muscles play a key role in physical activity and energy metabolism. The loss of skeletal muscle mass can cause problems related to metabolism and physical activity. Studies are being conducted to prevent such diseases by increasing the mass and regeneration capacity of muscles. Ginsenoside Rg5 has been reported to exhibit a broad range of pharmacological activities. However, studies on the effects of Rg5 on muscle differentiation and growth are scarce. Methods: To investigate the effects of Rg5 on myogenesis, C2C12 myoblasts were induced to differentiate with Rg5, followed by immunoblotting, immunostaining, and qRT-PCR for myogenic markers and promyogenic signaling (p38MAPK). Immunoprecipitation confirmed that Rg5 increased the interaction between MyoD and E2A via p38MAPK. To investigate the effects of Rg5 on prevention of muscle mass loss, C2C12 myotubes were treated with dexamethasone to induce muscle atrophy. Immunoblotting, immunostaining, and qRT-PCR were performed for myogenic markers, Akt/mTOR signaling for protein synthesis, and atrophy-related genes (Atrogin-1 and MuRF1). Results: Rg5 promoted C2C12 myoblast differentiation through phosphorylation of p38MAPK and MyoD/E2A heterodimerization. Furthermore, Rg5 stimulated C2C12 myotube hypertrophy via phosphorylation of Akt/mTOR. Phosphorylation of Akt induces FoxO3a phosphorylation, which reduces the expression of Atrogin-1 and MuRF1. Conclusion: This study provides an understanding of how Rg5 promotes myogenesis and hypertrophy and prevents dexamethasone-induced muscle atrophy. The study is the first, to the best of our knowledge, to show that Rg5 promotes muscle regeneration and to suggest that Rg5 can be used for therapeutic intervention of muscle weakness and atrophy, including cancer cachexia.