• 한국가금학회지
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• 제49권2호
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• pp.125-137
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• 2022

This study aimed to determine the effects of dietary microalgae (Tetradesmus sp. (TO)) on intestinal immunity and microbiota of pre-starter broilers. One hundred and twenty 1-day-old birds (Ross 308) were allocated to two dietary treatment groups with six blocks in a randomized complete block design. The two experimental diets consisted of a corn-soybean meal-based basal diet and a diet with 0.5% TO powder instead of cornstarch in the basal diet. After feeding the experimental diets for ten days, all birds' body weight and feed intake were measured, and representative eight birds were selected from each treatment group. Small intestinal lamina propria cells were isolated using flow cytometry to examine the frequency of immune cells. Cecal feces were harvested for 16s rRNA gut microbiota analysis and fecal IgA levels. Here, we found that 0.5% TO supplementation increased CD3⁺CD4⁺ T cells in the small intestine, but decreased CD3⁺CD8⁺ T cells in the small intestine. Gut microbial analysis showed that TO supplementation significantly increased the alpha diversity of the gut microbiome. Taxonomic analysis showed that TO treatment increased the abundance of Firmicutes and decreased that of Bacteroidetes at the phylum level. The distribution of Enterobacteriaceae containing many harmful bacteria at the family level, was lower in the TO group. In the LEfSe analysis, the TO group had a significantly enriched abundance of Agathobaculum at the genus level. Overall, results show that Tetradesmus sp. supplementation influences intestinal T-cell immunity and induces the expansion of beneficial gut microbes in pre-starter broiler chickens.

• Journal of Ginseng Research
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• 제46권5호
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• pp.628-635
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• 2022

Background: Ulcerative colitis (UC) is the large intestine disease that results in chronic inflammation and ulcers in the colon. Rg3-enriched Korean Red Ginseng extract (Rg3-RGE) is known for its pharmacological activities. Persicaria tinctoria (PT) is also used in the treatment of various inflammatory diseases. The aim of this study is to investigate the attenuating effects of Rg3-RGE with PT on oxazolone (OXA)-induced UC in mice. Methods: A total of six groups of mice including control group, OXA (as model group, 1.5%) group, sulfasalazine (75 mg/kg) group, Rg3-RGE (20 mg/kg) group, PT (300 mg/kg) group, and Rg3-RGE (10 mg/kg) with PT (150 mg/kg) group. Data on the colon length, body weight, disease activity index (DAI), histological changes, nitric oxide (NO) assay, Real-time PCR of inflammatory factors, ELISA of inflammatory factors, Western blot, and flow cytometry analysis were obtained. Results: Overall, the combination treatment of Rg3-RGE and PT significantly improved the colon length and body weight and decreased the DAI in mice compared with the treatment with OXA. Additionally, the histological injury was also reduced by the combination treatment. Moreover, the NO production level and inflammatory mediators and cytokines were significantly downregulated in the Rg3-RGE with the PT group compared with the model group. Also, NLR family pyrin domain containing 3 (NLRP3) inflammasome and nuclear factor kappa B (NF-𝛋B) were suppressed in the combination treatment group compared with the OXA group. Furthermore, the number of immune cell subtypes of CD4⁺ T-helper cells, CD19⁺ B-cells, and CD4⁺ and CD25⁺ regulatory T-cells (Tregs) was improved in the Rg3-RGE with the PT group compared with the OXA group. Conclusion: Overall, the mixture of Rg3-RGE and PT is an effective therapeutic treatment for UC.

• 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.

• 공업화학
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• 제33권1호
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• pp.83-89
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• 2022

현재 진행 중인 코로나19의 세계적 유행을 기점으로 유전자 전달을 통한 면역 형성에 대한 연구가 활발히 진행되고 있다. 특히 바이러스를 통한 유전자 전달이 부작용이 다수 발견됨에 따라 비바이러스성 유전자 전달체에 대한 요구가 크게 증가하였다. 본 연구에서는 생체적합물질인 히알루론 아민으로 코팅한 폴리에틸렌이민-플라스미드 DNA 복합체를 통한 효율적인 유전자 전달 시스템을 제안했다. 다양한 조성에서 생성된 폴리에틸렌이민-플라스미드 DNA 복합체(polyplex)와 히알루론 아민으로 코팅한 폴리에틸렌이민-플라스미드 DNA 복합체(polyplex-HA)의 크기 및 플라스미드 DNA 발현 정도를 비교해 각 물질의 최적 비율을 찾아냈고 복합체의 크기 및 제타 전위, 에너지 필터링 투과 전자현미경(EF-TEM) 이미지를 통해 입자의 특성을 평가했다. 세포 내 전달 및 발현 효율을 형광현미경과 유세포분석기를 통해 상용화 되어있는 유전자 전달체인 lipofectamine과 비교 분석했다. 본 연구에서 제안된 polyplex-HA는 pDNA 뿐만 아니라 다양한 유전물질을 전달할 수 있으며, 전달체에 대한 면역반응이 적어 다회성 투여에 유리하여 미래의 백신 플랫폼의 기반이 될 수 있을 것으로 기대할 수 있다.

• 생명과학회지
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• 제32권1호
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• pp.36-43
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• 2022

점막면역체계는 병원체에 대항하여 정밀하게 설계된 생물체의 방어체계로 그 중 위장관은 최전방에서 가장 중요한 기능을 하는 기관이다. 숙주 감염을 일으키는 병원성 미생물 가운데 식중독을 유발하는 살모넬라는 구강경로로 체내로 들어와 파이어판을 통해서 침입한다. 다양한 혈청형의 살모넬라 균주는 여러 톨유사수용체의 리간드를 통해서 숙주의 선천면역을 자극하지만, 숙주 특이성과 병원성에 따라 장내 점막에서 다양한 후천면역반응을 유도하기도 한다. 우리는 살모넬라 균주 중 비교적 연구가 덜 되어있는 S. enteritidis에 의해 감염되었을 때 장 내에서 일어나는 면역반응을 연구하였다. 우선 마우스에 처리할 적정 농도의 살모넬라 농도를 정하기 위해 살모넬라의 농도를 다르게 하여 실험을 진행하였고 고농도의 살모넬라 처리군에서 더 낮은 생존율과 높은 몸무게 감소율을 관찰하였다. 반수치사량의 반에 해당하는 농도의 살모넬라를 마우스에 감염시켰을 때 장 내 수지상세포와 T 세포에 유의적인 차이가 없는 것을 확인하였다. 하지만 동일한 조건의 마우스에서 회장조직을 염색했을 때 뮤신 분비의 증가와 술잔세포의 감소를 확인할 수 있었다. 또한 밀착연접단백질 유전자의 상대적인 발현량을 분석하였고 살모넬라 처리군에서 Claudin의 유의적인 감소를 확인할 수 있었다. 이어 살모넬라 감염 후 DSS를 처리한 실험에서 살모넬라 처리군에서 생존율이 낮아짐을 확인하여 S. enteritidis가 염증반응을 악화시킨다는 것을 알 수 있었다.

• 대한본초학회지
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• 제36권3호
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• pp.47-53
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• 2021

Objectives : Currently, the alopecia is one of the most emotionally stressful syndromes in human life. Human hair dermal papilla cells (HDPCs) play an essential role in controlling hair growth and in regulating hair cycle. We performed MTT assay, cell cycle, and western blot to determine the effects of essential oil from Coicis Semen (ECS) on hair growth in HDPCs. Methods : We monitored cell proliferations by MTT assay in HDPCs. After setting up the safe and effective concentration range to be treated ECS, cell cycle analysis was performed using flow cytometry. Also, the protein expression of hair growth-related factors such as insulin like growth factor-1 (IGF-1), Wnt, extracellular signal-regulated kinase (ERK), serine/threonine-specific protein kinase (Akt) in HDPCs was determined by western blot. Results : As results, cell proliferation was increased in ECS group compared to dimethyl sulfoxide (DMSO) group and minoxidil (MNXD) group. Cell number of ECS group was more decrease in sub G1 phase than cell number of DMSO group. Also, cell number of ECS group increased compared to cell number of DMSO group in G1 phase. Protein expression of ECS group was higher than protein expression of DMSO group on related hair growth factors (IGF-1, Wnt, ERK, Akt). Conclusion : As mentioned above, ECS increased cell proliferation and the protein expression of IGF-1, Wnt, ERK, and Akt. These results suggest that ECS could be used as a potential material for the treatment of alopecia by increasing the proliferation of HDPCs.

• Animal Bioscience
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• 제35권5호
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• pp.763-777
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• 2022

Objective: Excessive lipid accumulation in adipocytes results in prevalence of obesity and metabolic syndrome. Curcumin (CUR), a naturally phenolic active ingredient, has been shown to have lipid-lowering effects. However, its underlying mechanisms have remained largely unknown. Therefore, the study aims to determine the effect of CUR on cellular lipid accumulation in porcine subcutaneous preadipocytes (PSPA) and to clarify novel mechanisms. Methods: The PSPA were cultured and treated with or without CUR. Both cell counting Kit-8 and lactate dehydrogenase release assays were used to examine cytotoxicity. Intracellular lipid contents were measured by oil-red-o staining extraction and triglyceride quantification. Apoptosis was determined by flow cytometry and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling assay. Adipogenic and apoptosis genes were analyzed by quantitative polymerase chain reaction and Western blot. Results: The CUR dose-dependently reduced the proliferation and lipid accumulation of PSPA. Noncytotoxic doses of CUR (10 to 20 μM) significantly inhibited extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation and expression of adipogenic genes peroxisome proliferation-activity receptor-γ (PPAR-γ), CCAAT/enhancer binding protein-α, sterol regulatory element-binding protein-1c, adipocyte protein-2, glucose transporter-4 as well as key lipogenic enzymes fatty acid synthase and acetyl-CoA carboxylase, while ERK1/2 activation significantly reversed CUR-reduced lipid accumulation by increasing PPAR-γ. Furthermore, compared with differentiation induced media treated cells, higher dose of CUR (30 μM) significantly decreased the expression of AKT and B-cell lymphoma-2 (BCL-2), while increased the expression of BCL-2-associated X (BAX) and the BAX/BCL-2 expression ratio, suggesting triggered apoptosis by inactivating AKT and increasing BAX/BCL-2 ratio and Caspase-3 expression. Moreover, AKT activation significantly rescued CUR inhibiting lipid accumulation via repressing apoptosis. Conclusion: These results demonstrate that CUR is capable of suppressing differentiation by inhibiting ERK1/2-PPAR-γ signaling pathway and triggering apoptosis via decreasing AKT and subsequently increasing BAX/BCL-2 ratio and Caspase-3, suggesting that CUR provides an important method for the reduction of porcine body fat, as well as the prevention and treatment of human obesity.

• Journal of Ginseng Research
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• 제47권1호
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• pp.97-105
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• 2023

Background: Hyperactivated airway mucosa cells overproduce mucin and cause severe breathing complications. Here, we aimed to identify the effects of saponins derived from Panax ginseng on inflammation and mucin overproduction. Methods: NCI-H292 cells were pre-incubated with 16 saponins derived from P. ginseng, and mucin overproduction was induced by treatment with phorbol 12-myristate 13-acetate (PMA). Mucin protein MUC5AC was quantified by enzyme-linked immunosorbent assay, and mRNA levels were analyzed using quantitative polymerase chain reaction (qPCR). Moreover, we performed a transcriptome analysis of PMA-treated NCI-H292 cells in the absence or presence of Rg5, and differential gene expression was confirmed using qPCR. Phosphorylation levels of signaling molecules, and the abundance of lipid droplets, were measured by western blotting, flow cytometry, and confocal microscopy. Results: Ginsenoside Rg5 effectively reduced MUC5AC secretion and decreased MUC5AC mRNA levels. A systematic functional network analysis revealed that Rg5 upregulated cholesterol and glycerolipid metabolism, resulting in the production of lipid droplets to clear reactive oxygen species (ROS), and modulated the mitogen-activated protein kinase and nuclear factor (NF)-kB signaling pathways to regulate inflammatory responses. Rg5 induced the accumulation of lipid droplets and decreased cellular ROS levels, and N-acetyl-ⳑ-cysteine, a ROS inhibitor, reduced MUC5AC secretion via Rg5. Furthermore, Rg5 hampered the phosphorylation of extracellular signal-regulated kinase and p38 proteins, affecting the NF-kB signaling pathway and pro-inflammatory responses. Conclusion: Rg5 alleviated inflammatory responses by reducing mucin secretion and promoting lipid droplet-mediated ROS clearance. Therefore, Rg5 may have potential as a therapeutic agent to alleviate respiratory disorders caused by hyperactivation of mucosa cells.

• 생명과학회지
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• 제33권5호
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• pp.397-405
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• 2023

항산화제로서 산화적 손상의 방지에 중요한 역할을 하는 것으로 알려진 glutathione (GSH)의 면역 조절에 대한 연구는 현재까지 제대로 이루어지지 않았다. 본 연구에서 우리는 환원형 GSH인 luthione^®이 RAW 264.7 세포에서 면역 강화 효과가 있는지를 조사하였다. 유세포 분석 및 면역 형광 실험의 결과에 의하면, luthione은 대조군 세포에 비해 대식세포의 대표적인 기능인 식세포 활성을 luthione 처리 농도 의적으로 증가시키는 것으로 나타났다. 또한, cytokine array의 결과에 의하면, IL-5, IL-1β와 IL-27의 발현이 luthione이 처리된 세포에서 유의하게 증가하였다. 아울러 luthione에 의한 TNF-α 및 IL-1β의 생성 증가는 그들의 단백질 발현 증가를 통해 이루어졌으며, NO 및 PGE₂와 같은 면역 매개체 유리의 증가는 iNOS 및 COX-2의 발현 증가와 관련이 있었으며, 이는 M1 대식세포 분화 마커인 CD86 발현의 증가와 연관성이 있었다. 그리고 heatmap 분석을 통하여 SOCS1/3 매개 STAT/JAK 신호 전달 경로가 luthione에 의한 면역 조절 증가에 관여함을 확인하였다. 결론적으로, 우리의 결과는 luthione이 M1 macrophage polarization의 분자 조절자로 작용하여 면역 능력을 향상시킬 수 있음을 시사한다.

• The Korean Journal of Physiology and Pharmacology
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• 제27권5호
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• pp.457-470
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• 2023

The aim of this study was to investigate the role of kinesin superfamily member 15 (KIF15) in nasopharyngeal carcinogenesis (NPC) and explore its underlying mechanisms. We employed various assays, including the CCK-8 assay, flow cytometry, the Transwell and scratch assay, Western blotting, and nude mice transplantation tumor, to investigate the impact of KIF15 on NPC. Our findings demonstrate that KIF15 plays a critical role in the proliferation, apoptosis, migration, and invasion of NPC cells. Furthermore, we discovered that silencing KIF15 inhibits cell proliferation, migration, and invasion while promoting apoptosis, and that KIF15's effect on NPC cell growth is mediated through the PI3K/AKT and P53 signaling pathways. Additionally, we showed that KIF15 promotes nasopharyngeal cancer cell growth in vivo. Our study sheds light on the significance of KIF15 in NPC by revealing that KIF15 knockdown inhibits NPC cell growth through the regulation of AKT-related signaling pathways. These findings suggest that KIF15 represents a promising therapeutic target for the prevention and treatment of NPC.