• Animal Bioscience
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• 제36권2호
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• pp.200-208
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• 2023

Objective: Muscle acetylcholine receptors have five alpha subunits (α, β, δ, ε, or γ), and cholinergic receptor nicotinic gamma subunit (CHRNG) is the γ subunit. It may also play an essential role in biological processes, including cell differentiation, growth, and survival, while the role of CHRNG has not been studied in the literature. Therefore, the purpose of this study is to clarify the effect of CHRNG on the proliferation and differentiation of bovine preadipocytes. Methods: We constructed a CHRNG overexpression adenovirus vector and successfully overexpressed it on bovine preadipocytes. The effects of CHRNG on bovine preadipocyte proliferation were detected by Edu assay, cell counting Kit-8 (CCK-8), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), Western blot and other techniques. We also performed oil red O, RT-qPCR, Western blot to explore its effect on the differentiation of preadipocytes. Results: The results of Edu proliferation experiments showed that the number of EDU-positive cells in the overexpression group was significantly less. CCK-8 experiments found that the optical density values of the cells in the overexpression group were lower than those of the control group, the mRNA levels of proliferating cell nuclear antigen (PCNA), cyclin A2 (CCNA2), cyclin B1 (CCNB1), cyclin D2 (CCND2) decreased significantly after CHRNG gene overexpression, the mRNA levels of cyclin dependent kinase inhibitor 1A (CDKN1A) increased significantly, and the protein levels of PCNA, CCNB1, CCND2 decreased significantly. Overexpression of CHRNG inhibited the differentiation of bovine preadipocytes. The results of oil red O and triglyceride determination showed that the size and speed of lipid droplets accumulation in the overexpression group were significantly lower. The mRNA and protein levels of peroxisome proliferator activated receptor gamma (PPAR class="checkNonKBPoint">γ), CCAAT enhancer binding protein alpha (CEBPα), fatty acid binding protein 4 (FABP4), fatty acid synthase (FASN) decreased significantly. Conclusion: Overexpression of CHRNG in bovine preadipocytes inhibits the proliferation and differentiation of bovine preadipocytes.

• Animal cells and systems
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• 제13권2호
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• pp.235-245
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• 2009

Embryonic stem (ES) cells have a capability to generate all types of cells. However, the mechanism by which ES cells differentiate into specific cell is still unclear. Using microarray technology, the differentiation process in mouse embryonic stem cells was characterized by temporal gene expression changes of mouse ES cells during differentiation in a monolayer culture. A large number of genes were differentially regulated from 1 day to 14 days, and less number of genes were differentially expressed from 14 days to 28 days. The number of up-regulated genes was linearly increased throughout the 28 days of in vitro differentiation, while the number of down-regulated genes reached the plateau from 14 days to 28 days. Most differentially expressed genes were functionally classified into transcriptional regulation, development, extra cellular matrix (ECM),cytoskeleton organization, cytokines, receptors, RNA processing, DNA replication, chromatin assembly, proliferation and apoptosis related genes. While genes encoding ECM proteins were up-regulated, most of the genes related to proliferation, chromatin assembly, DNA replication, RNA processing, and cytoskeleton organization were down-regulated at 14 days. Genes known to be associated with embryo development or transcriptional regulation were differentially expressed mostly after 14 days of differentiation. These results indicate that the altered expression of ECM genes constitute an early event during the spontaneous differentiation, followed by the inhibition of proliferation and lineage specification. Our study might identify useful time-points for applying selective treatments for directed differentiation of mouse ES cells.

• Journal of Animal Science and Technology
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• 제62권6호
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• pp.854-863
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• 2020

This study was aimed to investigate the inhibitory effects of Porphyra dentata (P. dentata) extract on the adipogenesis of 3T3-L1 cells and evaluate its anti-obesity effect. The proliferation of 3T3-L1 cells and differentiation of adipocytes under treatment of P. dentata extract was examined by measuring the cell viability using alamarBlue assay and lipid droplets by Oil Red O staining. Results showed that P. dentata extract has no cytotoxicity effect and lipid droplets formation decreased in a concentration-dependent manner in 3T3-L1 cells. It has been confirmed that transcription factors affecting lipid accumulation and anti-adipogenic effects during cell differentiation are linked to P. dentata extract. We observed that P. dentata shows lowering the mRNA expression of peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα) that adipogenesis-associated key transcription factors and inhibiting adipogenesis in the early stages of differentiation. Treating the cells with P. dentata did not only suppressed PPARγ2 and C/EBPα but also significantly decreased the mRNA expression of adiponectin, Leptin, fatty acid synthase, adipocyte protein 2, and Acetyl-coA carboxylase 1. Overall, the P. dentata extract demonstrated inhibitory property in adipogenesis, which has a potential effect in anti-obesity in 3T3-L1 cells.

• Natural Product Sciences
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• 제14권2호
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• pp.113-117
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• 2008

Saururus chinensis is a commonly used folk herb for the treatment of edema and liver diseases in Korea. To study the biological activity of Saururus chinensis in bone metabolism, we evaluated the effect of its extracts on osteoclast differentiation in vitro using primary mouse bone marrow-derived macrophages. Methanol extract (ME) from dried roots of Saururus chinensis was partitioned into methylene chloride (MF), ethyl acetate (EF), n-butanol (BF) and water fractions (WF). Tartrate-resistance acid phosphatase (TRAP) activity assay and western blot analysis were performed to determine the effect on osteoclast differentiation and mitogen-activated protein (MAP) kinases activation. ME, MF and EF dramatically inhibited receptor activator of ${NF-kB}$ ligand (RANKL)-induced formation of multinucleated osteoclasts and activation of MAP kinases. This study firstly demonstrated that ME, MF and EF of Saururus chinensis have the potential to inhibit the osteoclast differentiation, which results from the inhibition of MAP kinases activations in part.

• Journal of Animal Science and Technology
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• 제49권5호
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• pp.593-598
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• 2007

본 연구는 스테로이드 성호르몬 즉 에스트로겐, 테스토스테론 및 노르테스토스테론이 돼지 지방전구세포의 분화와 증식에 미치는 영향을 구명하기 위해서 수행하였다. 지방전구세포는 갓난 암퇘지의 등지방조직을 떼어내어 coll- agenase를 처리해서 분리해서 배양했다. 세포 배양중에 10－8M과 10－7M의 스테로이드 성호르몬을 처리했다. 세포배양 전기에 스테로이드 성호르몬을 처리했을 때 지방전구세포의 분화나 증식에 아무런 영향을 미치지 않았고, 세포배양 후기에 처리했을 때는 테스토스테론과 노르테스토스테론이 세포분화를 촉진시켰다.

• Journal of Animal Science and Technology
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• 제49권1호
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• pp.25-32
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• 2007

본 연구는 여러 CLA 이성체, 즉 cis-9, cis- 11(c9c11), cis-9, trans-11(c9t11), trans-9, trans-11 (t9t11) 및 trans-10, cis-12(t10c12)가 배양 중인 돼지 지방세포와 근육세포의 분화와 증식에 미치는 영향을 구명하고자 실시하였다. 세포는 신생자돈으로부터 분리했다. t10c12 이성체는 지방세포의 분화를 억제했는데(92%) 근육세포의 분화는 억제시키지 않았다. t9t11 이성체는 지방세포의 분화를 억제했는데(14%), 근육세포의 분화는 촉진시켰다(26%). 다른 CLA 이성체는 지방세포나 근육세포의 분화에 영향을 미치지 않았다. 그리고 CLA가 지방세포와 근육세포의 증식에 미치는 영향은 분화에 미치는 영향에 비해서 작았다. 위의 결과는 여러 CLA 이성체는 돼지 지방세포와 근육세포의 분화에 다른 영향을 미침을 나타낸다.

• Journal of Animal Science and Technology
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• 제46권6호
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• pp.967-974
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• 2004

본 연구는 여 러 conjugated linoleic acid(CLA) 이성체가 돼지 지방전구세포의 분화에 미치는 영향을 구영하기 위해 수행하였다. 돼지 지방전구세포는 갓난 돼지의 등 지방에서 분리해서 성숙지방세포로 분화될 때 까지 배양했다. 여러 CLA 이성체를 배양중의 세포에 처리했다. 세포분화는 세포배양이 끝난 후 세포의 glycerol-3-phosphate의 활성도를 측정함으로써 구명했다. 20$\mu$M과 50$\mu$M의 transto-cis12 CLA 이성체는 돼지 지방천구세포의 분화를 억제했고，한펀 cis9-cisl1 이성체는 세포분화를 촉진했다. cis9-trans 11 과 trans9-trans11 이생체는 세포분화에 아무런 영향을 마치지 않았다. CLA의 세포분화에 미치는 작용은 배양후기 (day8${\sim}$14) 보다 배양전기(day 0${\sim}$ 8)에 더 두드러지게 나타냈다. 위의 결과는 여러 CLA 이성체는 돼지 지방천구세포 분화에 각각 다른 작용을 가짐을 나타낸다.

• Molecules and Cells
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• 제20권1호
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• pp.57-68
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• 2005

Murine erythroleukemia (MEL) cells are widely used to study erythroid differentiation thanks to their ability to terminally differentiate in vitro in response to chemical induction. At the molecular level, not much is known of their terminal differentiation apart from activation of adult-type globin gene expression. We examined changes in gene expression during the terminal differentiation of these cells using microarray-based technology. We identified 180 genes whose expression changed significantly during differentiation. The microarray data were analyzed by hierarchical and k-means clustering and confirmed by semi-quantitative RT-PCR. We identified several genes including H1f0, Bnip3, Mgl2, ST7L, and Cbll1 that could be useful markers for erythropoiesis. These genetic markers should be a valuable resource both as potential regulators in functional studies of erythroid differentiation, and as straightforward cell type markers.

• 한국한의학연구원논문집
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• 제13권1호통권19호
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• pp.139-145
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• 2007

Emodin is one of the main active components of a traditional Korean medicine isolated from the root and rhizomes of Rheum palmatum L. In this study, of 222 natural compounds to evaluate the anabolic activities, emodin activated bone morphogenetic protein (BMP)-2 promoter in the differentiation process of mouse osteoblastic MC3T3-E1 cells. Emodin was shown to significantly stimulate the activity and expression of alkaline phosphatase, an earlyphase marker of osteoblastic differentiation, on the differentiation day 7, and induce the osteopontin mRNA expression from the differentiation day 14. In addition, low concentration (up to 5 M) of emodin dramatically promoted the induction of mineralization in MC3T3-E1 subclone 4 cells. The stimulatory effect of emodin on the osteoblast differentiation/mineralization could be associated with its potential to stimulate the BMP-2 gene expression. Although further studies are needed to determine the precise mechanism, this study suggests that the use of herbal medicine containing natural compounds with anabolic activity such as emodin could have a beneficial effect on bone health.

• 원예과학기술지
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• 제33권1호
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• pp.31-38
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• 2015

This study was carried out to investigate the relationship between endogenous polyamines (PAs) and floral bud differentiation in chrysanthemum (Chrysanthemum morifolium). In this study, PA content (free, bound, and conjugated) in apical buds, leaves, and roots changed appreciably during floral bud differentiation. PAs accumulated during series of processes such as floral induction, differentiation of floret primordia, and crown formation in apical buds; changes in PAs in apical buds may have a relationship with those in leaves and roots. The levels of free PAs and conjugated PAs [putrescine (Put) and spermine (Spm)] in apical buds rapidly increased during the initiation stage of floral bud differentiation, while free and conjugated spermidine (Spd) reached their highest levels at the stage of floret primordium differentiation. In the free, conjugated, and bound PA fractions, the changes in Spm content were negligible compared to those of Put and Spd throughout the experiment. These findings indicate that PAs participate in regulating the process of flower bud differentiation in chrysanthemum.