• Animal Bioscience
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• 제34권4호
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• pp.546-557
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• 2021

Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.

• 한국자원식물학회지
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• 제35권2호
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• pp.372-379
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• 2022

본 연구는 추출용매에 따른 레몬밤 추출물의 중성지방 조절에 대한 논문으로 3T3-L1 지방전구세포를 이용해 지방세포로 분화유도한 뒤 lipid accumulation, triglyceride contents, PKA, HSL, perilipin, ATGL 및 CGI-58 단백질 발현을 확인하였다. 실험결과 네가지 조건의 레몬밤 추출물 중 물 추출 조건인 MOW100 추출물만 lipid accumulation과 triglyceride contents의 유의적 억제 효능을 나타내었으며, MOE70, MOE50, MOE50 추출물은 lipid accumulation과 triglyceride contents의 감소 효과가 없거나 유의적 차이를 나타내지 않는 것이 확인되었다. MOW100 추출물의 세포내 지방 축적 억제는 지방세포 내 lipolysis를 조절하는 효소 조절을 통해 나타난 결과로 판단된다. 따라서 레몬밤 물 추출물은 cAMP-PKA를 활성화시키고, lipolysis를 조절하는 효소의 상승작용을 통해 세포내 중성지방을 조절함으로써 혈중 중성지방을 개선하는 후보소재로의 사용이 가능할 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제26권6호
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• pp.541-556
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• 2022

Myocardial fibrosis is a key link in the occurrence and development of diabetic cardiomyopathy. Its etiology is complex, and the effect of drugs is not good. Cardiomyocyte apoptosis is an important cause of myocardial fibrosis. The purpose of this study was to investigate the effect of gaseous signal molecule sulfur dioxide (SO₂) on diabetic myocardial fibrosis and its internal regulatory mechanism. Masson and TUNEL staining, Western-blot, transmission electron microscopy, RT-qPCR, immunofluorescence staining, and flow cytometry were used in the study, and the interstitial collagen deposition, autophagy, apoptosis, and changes in phosphatidylinositol 3-kinase (PI3K)/AKT pathways were evaluated from in vivo and in vitro experiments. The results showed that diabetic myocardial fibrosis was accompanied by cardiomyocyte apoptosis and down-regulation of endogenous SO₂-producing enzyme aspartate aminotransferase (AAT)_1/2. However, exogenous SO₂ donors could up-regulate AAT_1/2, reduce apoptosis of cardiomyocytes induced by diabetic rats or high glucose, inhibit phosphorylation of PI3K/AKT protein, up-regulate autophagy, and reduce interstitial collagen deposition. In conclusion, the results of this study suggest that the gaseous signal molecule SO₂ can inhibit the PI3K/AKT pathway to promote cytoprotective autophagy and inhibit cardiomyocyte apoptosis to improve myocardial fibrosis in diabetic rats. The results of this study are expected to provide new targets and intervention strategies for the prevention and treatment of diabetic cardiomyopathy.

• 한국응용곤충학회지
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• 제60권2호
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• pp.193-199
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• 2021

벌목 개미과의 일본열마디개미는 국내 토착종이다. 지금까지 야외에서 이 종의 월동기작을 이해하기 위한 내한성 연구는 진행되지 않았다. 저온에서 다양한 온도별 노출시간에 따른 발육태별 저온 저항성을 조사하였다. 성충인 일개미가 다른 발육태와 비교하여 5℃와 10℃의 저온에서 높은 생존율을 보였으며, 급속내한성 유기 조건인 15℃에서 12시간 노출 후 내한성을 획득하였다. 급속내한성 유기는 10℃에서 최대 44%까지 생존율이 향상되었으며, 체내과냉각점과 체내빙점은 각각 -10.0℃에서 -14.2℃, -11.3℃에서 -15.3℃까지 낮아졌다. 저온처리는 저온 또는 스트레스 관련 유전자인 글리세롤 인산화효소와 열충격 단백질의 발현을 증가시켰다. 이상의 결과는 일본열마디개미의 내한성이 급속내한성 유기에 의해 야기된다는 것을 의미한다.

• 대한한의학방제학회지
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• 제30권2호
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• pp.85-93
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• 2022

Objectives : This study investigated anti-inflammatory effects of Nardotidis seu Sulculii Concha water extract (NSCE) against restraint-induced stress. Methods : In vivo, NSCE was orally administered to male white mice at concentrations of 250 mg/kg and 500 mg/kg for 3 days, and then restraint-induced stress was induced for 6 hours. The level of liver damage was measured by serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). The stress-related hormones such as cortisol and corticosterone were measured by ELISA assay. Also, western blot analysis was performed to detect expression of mitogen-activated protein kinase (MAPK) and cyclooxygenase-2 (COX-2) proteins. Pathological changes were observed by hematoxylin and eosin (H&E) staining of the liver tissue, and Immunohistochemical (IHC) staining was performed to examine liver inflammation through macrophage infiltration. Results : The AST, ALT, LDH and the stress related hormones such as cortisol and corticosterone were significantly decreased in the NSCE treated group compared with stress group. In histological analysis, H&E staining of liver tissues did not detect the hepatic injury or damage in all groups. As a result of IHC staining, it was confirmed that infiltration of macrophages was increased in the stress-induced group, but decreased in the group treated with NSCE. The COX-2 and MAPK proteins expression was significantly increased by restraint-induced stress, but these proteins were decreased in the NSCE treated group. Conclusions : These results suggest that NSCE has the anti-inflammatory activity in restraint-induced stress model, and it is believed that NSCE can be used for the prevention of liver inflammation.

• 대한본초학회지
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• 제33권4호
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• pp.101-108
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• 2018

Objectives : This study aimed to effects antioxidant activity of citrus peel extract (CPE) and effect on its glucose metabolism in L6 rat skeletal muscle cells. Methods : Antioxidative activities were evaluated by using 10 kinds of natural materials, and total polyphenol and flavonoid contents were examined. The L6 muscle cells toxicity of CPE was examined by MTT assay. Expression of glucose-related genes in L6 muscle cells by CPE treatment was analyzed by real-time PCR and western blotting. Results : The $IC_{50}$ values of DPPH and ABTS free radical scavenging activity of CPE were ($15.47{\pm}0.26{\mu}g/m{\ell}$ and $12.07{\pm}1.23{\mu}g/m{\ell}$, respectively), effectively clearing DPPH and ABTS. CPE showed total polyphenol and flavonoid contents ($20.30{\pm}0.38$ and $64.20{\pm}0.52$, respectively). The selected CPE were used in experiments using an effective concentration that is not toxic in L6 muscle cells. We investigated insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3-kinase regulatory (PI3KR), Akt, and glucose transporter 4 (GLUT4). mRNA analysis by realtime PCR showed no significant difference, but CPE-treated cells showed a tendency to increase in concentration-dependent manner. However, analysis of protein expression of Akt and GLUT4 by western blotting showed that CPE treatment significantly increased concentration dependent (p<0.001). Conclusions : As a result, citrus peel extract with high antioxidant activity regulates glucose metabolism in L6 muscle cells. Therefore, CPE can be a potential treatment for the treatment of diabetes.

• Journal of Applied Biological Chemistry
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• 제65권3호
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• pp.203-213
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• 2022

미선나무(Abeliophyllum distichum) 1속 1종의 한반도 고유식물이다. 꽃잎색의 변이에 따라 미선(백색), 분홍미선, 상아미선 그리고 연노랑색의 옥황1호 품종 등으로 구분된다. 본 연구에서는 4종류의 꽃 추출물(FAD)로부터 생리활성물질의 함량과 항염증 활성 차이를 비교·분석하였다. FAD의 생리활성 물질은 LC/MS 분석을 통해 rutin, hirsutrin, 그리고 acteoside를 분석하였다. DPPH와 ABTS 라디칼 소거활성을 통한 항산화 활성을 분석하였으며, 항산화 활성은 각 추출물의 생리활성 물질의 함량에 따라 유의성 있는 활성을 나타내었다. FAD는 LPS로 유도된 RAW 264.7 세포에서 염증 유발 매개체(산화질소, iNOS, COX-2)의 발현을 감소시켰다. 또한 NF-κB와 MAPK 신호전달 경로 조절을 통한 항염증 효과를 확인하였다.

• Animal Bioscience
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• 제35권3호
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• pp.367-376
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• 2022

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry as well as the economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for HPAIV resistance. Therefore, in this study, we investigated gene expression related to cytokine-cytokine receptor interactions by comparing resistant and susceptible Ri chicken lines for avian influenza virus infection. Methods: Ri chickens of resistant (Mx/A; BF2/B21) and susceptible (Mx/G; BF2/B13) lines were selected by genotyping the Mx dynamin like GTPase (Mx) and major histocompatibility complex class I antigen BF2 genes. These chickens were then infected with influenza A virus subtype H5N1, and their lung tissues were collected for RNA sequencing. Results: In total, 972 differentially expressed genes (DEGs) were observed between resistant and susceptible Ri chickens, according to the gene ontology and Kyoto encyclopedia of genes and genomes pathways. In particular, DEGs associated with cytokine-cytokine receptor interactions were most abundant. The expression levels of cytokines (interleukin-1β [IL-1β], IL-6, IL-8, and IL-18), chemokines (C-C Motif chemokine ligand 4 [CCL4] and CCL17), interferons (IFN-γ), and IFN-stimulated genes (Mx1, CCL19, 2'-5'-oligoadenylate synthase-like, and protein kinase R) were higher in H5N1-resistant chickens than in H5N1-susceptible chickens. Conclusion: Resistant chickens show stronger immune responses and antiviral activity (cytokines, chemokines, and IFN-stimulated genes) than those of susceptible chickens against HPAIV infection.

• 한방안이비인후피부과학회지
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• 제35권4호
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• pp.75-94
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• 2022

Objectives : To identify the active ingredient of Poncirus Trifoliata Immaturus and to explore the mechanism expected to potentially act on dermatitis accompanied by pruritus. Methods : We conducted the network pharmacological analysis. We selected effective ingredients among the active compounds of Poinciri Fructus Immaturus. We found the target protein of the selected active ingredient, disease(dermatitis accompanied by pruritus) and fexofenadine. Then we established the network between the proteins which Poinciri Fructus Immaturus and fexofenadine intersected with disease respectively, and the coregene was also extracted. After that, the active pathways in the human body involving the groups and coregenes were searched. Results : Total of 7 active ingredients were selected, and 202 target proteins were collected. There were 756 proteins related to inflammatory skin disease accompanied by pruritus, and 75 proteins were related to fexofenadine. 42 proteins crossed by Poinciri Fructus Immaturus with a disease, and 31 proteins crossed by fexofenadine with a disease. 12 proteins were found as a coregene from the proteins that cross Poinciri Fructus Immaturus and disease. Coregenes are involved in 'Nitric-oxide synthase regulator activity', 'Epidermal growth factor receptor signaling pathway'. 2 groups that extracted are invloved in 'Fc receptor signaling pathway', 'Central carbon metabolism in cancer', 'Phosphatidylinositol 3-kinase complex, class IB', and 'omega-hydroxylase P450 pathway'. Conclusion : It is expected that Poinciri Fructus Immaturus will be able to show direct or indirect anti-pruritus and anti-inflammatory effects on skin inflammation accompanied by pruritus in the future. And it is also expected to have a synergy effect with fexofenadine on skin disease.

• Animal Bioscience
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• 제36권3호
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• pp.404-416
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• 2023

Objective: Daweizi (DWZ) is a famous indigenous pig breed in China and characterized by tender meat and high fat percentage. However, the expression profiles and functions of transcripts in DWZ pigs is still in infancy. The object of this study was to depict the transcript profiles in DWZ pigs and screen the potential pathway influence adipogenesis and fat deposition, Methods: Histological analysis of backfat tissue was firstly performed between DWZ and lean-type Yorkshire pigs, and then RNA sequencing technology was utilized to explore miRNAs, lncRNAs and mRNAs profiles in backfat tissue. 18 differentially expressed (DE) transcripts were randomly selected for quantitative real-time polymerase chain reaction (QPCR) to validate the reliability of the sequencing results. Finally, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were conducted to investigate the potential pathways influence adipocyte differentiation, adipogenesis and lipid metabolism, and a schematic model was further proposed. Results: A total of 1,625 differentially expressed transcripts were identified in DWZ pigs, including 27 upregulated and 45 downregulated miRNAs, 64 upregulated and 119 down-regulated lncRNA, 814 upregulated and 556 downregulated mRNAs. QPCR analysis exhibited strong consistency with the sequencing data. GO and KEGG analysis elucidated that the differentially expressed transcripts were mainly associated with cell growth and death, signal transduction, peroxisome proliferator-activated receptors (PPAR), AMP-activated protein kinase (AMPK), PI3K-Akt, adipocytokine and foxo signaling pathways, all of which are strongly involved in cell development, lipid metabolism and adipogenesis. Further analysis indicated that the BGIR9823_87926/miR-194a-5p/AQP7 network may be effective in the process of adipocyte differentiation or adipogenesis. Conclusion: Our study provides comprehensive insights into the regulatory network of backfat deposition and lipid metabolism in pigs from the point of view of miRNAs, lncRNAs and mRNAs.