• Journal of agriculture & life science
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• v.50 no.2
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• pp.139-150
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• 2016

Few studies have been reported that horse-bone extract(HBE) can prevent and treatment of bone diseases. However, HBE' therapeutic activities are still not fully understood. This study determined whether HBE up-regulates hemeoxygenase 1(HO-1) and this mediates its anti-inflammatory effect in murine macrophages.Nitric oxide(NO) assay, MTT assay and DPPH assay were performed. In addition, Western blotting and real time PCR were used to determine protein expression, and gene expression, respectively. HBE significantly inhibited NO production without observable cytotoxicity. In addition, HBE attenuated inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX-2) and phospho (p)-ERK protein expressions in LPS(0.1㎍/ml) stimulated RAW264.7 cells. On the other hand, HBE alone up-regulated HO-1 and Nrf-2 expressions, which mediated HBE's anti-inflammatory effect in RAW264.7 cells. Finally, HBE up-regulated HO-1 and impaired ERK1/2 signaling pathways, and thus it may provide protection against cellular oxidation and inflammation.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1213-1227
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• 2023

Fetal growth restriction (FGR) is a prevalent obstetric condition. This study aimed to investigate the role of Toll-like receptor 9 (TLR9) in regulating the inflammatory response and gut microbiota structure in FGR. An FGR animal model was established in rats, and ODN1668 and hydroxychloroquine (HCQ) were administered. Changes in gut microbiota structure were assessed using 16S rRNA sequencing, and fecal microbiota transplantation (FMT) was conducted. HTR-8/Svneo cells were treated with ODN1668 and HCQ to evaluate cell growth. Histopathological analysis was performed, and relative factor levels were measured. The results showed that FGR rats exhibited elevated levels of TLR9 and myeloid differentiating primary response gene 88 (MyD88). In vitro experiments demonstrated that TLR9 inhibited trophoblast cell proliferation and invasion. TLR9 upregulated lipopolysaccharide (LPS), LPS-binding protein (LBP), interleukin (IL)-1β and tumor necrosis factor (TNF)-α while downregulating IL-10. TLR9 activated the TARF3-TBK1-IRF3 signaling pathway. In vivo experiments showed HCQ reduced inflammation in FGR rats, and the relative cytokine expression followed a similar trend to that observed in vitro. TLR9 stimulated neutrophil activation. HCQ in FGR rats resulted in changes in the abundance of Eubacterium_coprostanoligenes_group at the family level and the abundance of Eubacterium_coprostanoligenes_group and Bacteroides at the genus level. TLR9 and associated inflammatory factors were correlated with Bacteroides, Prevotella, Streptococcus, and Prevotellaceae_Ga6A1_group. FMT from FGR rats interfered with the therapeutic effects of HCQ. In conclusion, our findings suggest that TLR9 regulates the inflammatory response and gut microbiota structure in FGR, providing new insights into the pathogenesis of FGR and suggesting potential therapeutic interventions.

• Journal of fish pathology
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• v.36 no.2
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• pp.323-336
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• 2023

This study was conducted to find out the effect of yeast by-products discarded after beer production as feed additives for carp (Cyprinus carpio). After producing feed by adding high-temperature dried beer yeast by-products (HD), freeze-dried beer yeast by-products (FD), and freeze-dried fermented beer yeast by-products (FF) after lactobacilli fermentation, innate immunity indicators, survival rates, and challenge experiments were evaluated. Both ACH50 and lysozyme activity were significantly increased (p<0.05) in the experimental group of FF 0.2% and 0.5% compared to the control group from day 7 to day 21. In addition, phagocyte activity was significantly increased (p<0.05) in the group of FF 0.5% compared to the control group at all time points. Both IL-1β and TNF-α expression levels increased significantly in the FD and FF groups on day 21 compared to the control group (p<0.05). In addition, the FF 0.5% group showed significantly higher expression levels (p<0.05) at all time points. Similarly, IL-10 expression increased significantly (p<0.05) in FF 0.2% and 0.5% groups at all time points. SOD gene expression was significantly increased in FD 0.5% and all FF groups on day 14 and 21 (p<0.05). The results of a 10-day challenge experiment using Edwardsiella piscicida (E. piscicida) showed a higher relative survival rate than the control group at all concentrations that fed FD and FF. In summary, it is estimated that 0.5% FF can effectively improve the innate immunity, growth rate, and antibacterial properties of carp rather than using discarded beer yeast supernatant alone as a functional feed additive.

• IMMUNE NETWORK
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• v.24 no.2
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• pp.3.1-3.23
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• 2024

Cigarette smoke extract (CSE)-treated mouse airway epithelial cells (MAECs)-derived exosomes accelerate the progression of chronic obstructive pulmonary disease (COPD) by upregulating triggering receptor expressed on myeloid cells 1 (TREM-1); however, the specific mechanism remains unclear. We aimed to explore the potential mechanisms of CSE-treated MAECs-derived exosomes on M1 macrophage polarization and pyroptosis in COPD. In vitro, exosomes were extracted from CSE-treated MAECs, followed by co-culture with macrophages. In vivo, mice exposed to cigarette smoke (CS) to induce COPD, followed by injection or/and intranasal instillation with oe-TREM-1 lentivirus. Lung function and pathological changes were evaluated. CD68⁺ cell number and the levels of iNOS, TNF-α, IL-1β (M1 macrophage marker), and pyroptosis-related proteins (NOD-like receptor family pyrin domain containing 3, apoptosis-associated speck-like protein containing a caspase-1 recruitment domain, caspase-1, cleaved-caspase-1, gasdermin D [GSDMD], and GSDMD-N) were examined. The expression of maternally expressed gene 3 (MEG3), spleen focus forming virus proviral integration oncogene (SPI1), methyltransferase 3 (METTL3), and TREM-1 was detected and the binding relationships among them were verified. MEG3 increased N6-methyladenosine methylation of TREM-1 by recruiting SPI1 to activate METTL3. Overexpression of TREM-1 or METTL3 negated the alleviative effects of MEG3 inhibition on M1 polarization and pyroptosis. In mice exposed to CS, EXO^-CSE further aggravated lung injury, M1 polarization, and pyroptosis, which were reversed by MEG3 inhibition. TREM-1 overexpression negated the palliative effects of MEG3 inhibition on COPD mouse lung injury. Collectively, CSE-treated MAECs-derived exosomal long non-coding RNA MEG3 may expedite M1 macrophage polarization and pyroptosis in COPD via the SPI1/METTL3/TREM-1 axis.

• IMMUNE NETWORK
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• v.23 no.6
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• pp.48.1-48.21
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• 2023

Mesenchymal stromal/stem cells (MSCs) possess immunoregulatory properties and their regulatory functions represent a potential therapy for acute lung injury (ALI). However, uncertainties remain with respect to defining MSCs-derived immunomodulatory pathways. Therefore, this study aimed to investigate the mechanism underlying the enhanced effect of human recombinant bone morphogenic protein-2 (rhBMP-2) primed ES-MSCs (MSC^BMP2) in promoting Tregs in ALI mice. MSC were preconditioned with 100 ng/ml rhBMP-2 for 24 h, and then administrated to mice by intravenous injection after intratracheal injection of 1 mg/kg LPS. Treating MSCs with rhBMP-2 significantly increased cellular proliferation and migration, and cytokines array reveled that cytokines release by MSC^BMP2 were associated with migration and growth. MSC^BMP2 ameliorated LPS induced lung injury and reduced myeloperoxidase activity and permeability in mice exposed to LPS. Levels of inducible nitric oxide synthase were decreased while levels of total glutathione and superoxide dismutase activity were further increased via inhibition of phosphorylated STAT1 in ALI mice treated with MSC^BMP2. MSC^BMP2 treatment increased the protein level of IDO1, indicating an increase in Treg cells, and Foxp3⁺CD25⁺ Treg of CD4⁺ cells were further increased in ALI mice treated with MSC^BMP2. In co-culture assays with MSCs and RAW264.7 cells, the protein level of IDO1 was further induced in MSC^BMP2. Additionally, cytokine release of IL-10 was enhanced while both IL-6 and TNF-α were further inhibited. In conclusion, these findings suggest that MSC^BMP2 has therapeutic potential to reduce massive inflammation of respiratory diseases by promoting Treg cells.

• The Journal of Korean Obstetrics and Gynecology
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• v.37 no.1
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• pp.1-22
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• 2024

목 적: 본 연구의 목적은 유방암 세포(MCF-7) 이식 누드 마우스 모델을 이용하여 육종용 열수 추출물의 면역활성 효과를 통한 항암 활성을 체계적으로 평가하는 것이다. 육종용 열수 추출물은 유방암 치료제로 자주 사용되는 대표적인 경구용 항암제인 tamoxifen 경구 투여군과 비교하여 분석 연구하였다. 방 법: 총 110마리의 6주령 암컷 누드 마우스를 준비하여, 7일간 적응 후 체중이 일정한 마우스를 선정하여 우측 둔부 피하부위에 MCF-7 세포를 이식하였다. 종양세포 이식을 한 지 20일 후, 종양 크기 및 체중을 기준으로 그룹 당 8마리씩 본 실험에 사용하였다. MCF-7 이종 이식 21일 후부터 매일 1회씩 35일간 육종용 열수 추출물을 10 ml/kg의 용량(400, 200 및 100 mg/kg)으로 경구 투여하였으며, tamoxifen 역시 10 ml/kg의 용량(20 mg/kg)으로 경구 투여하였고, 정상 및 종양 이식 매체 대조군에서는 멸균증류수만 종양 이식 21일 후부터 동일한 방법으로 35일간 경구 투여 하였다. 결 과: 본 실험의 결과, MCF-7 세포 이식을 함으로써 현저한 비장 및 하악하 림프절 무게, 혈중 IFN-γ의 함량, IL-1β 및 IL-10의 함량, 비장내 TNF-α, 비장세포 및 복강 대식구의 활성의 감소가 관찰되었고, 비장 및 하악하 림프절의 림프구 감소에 의한 조직병리학적 위축 또한 관찰되었다. 그리고 체중 및 증체량의 감소 역시 관찰되었으며, 혈중 IL-6 함량의 증가, 난소 주위의 지방 무게의 감소 및 조직병리학적으로 난소 주위의 축적 지방 조직 위축 현상이 인정되어, 종양 이식 후에 전형적인 종양과 관련된 면역억제와 악액질 현상이 유발된 것으로 판단되었다. 한편 육종용 열수 추출물 400, 200 및 100 mg/kg 경구 투여군에서는 종양 이식 대조군에 비해 유의성 있는 현저한 항암활성이 투여 용량 의존적으로 관찰되었다. 또한 tamoxifen 20 mg/kg 경구 투여군에서는 종양 관련 악액질 소견이 오히려 악화되는 반면, 육종용 열수 추출물 경구 투여군에서는 면역활성 및 악액질 억제 효과가 관찰되었다. 결 론: 본 연구 결과, 육종용 열수 추출물의 적절한 경구 투여는 심각한 부작용 없이, 종양 관련 악액질 소견을 포함하여, 효과적인 유방암 치료 수단을 제공할 수 있을 것으로 기대된다.

• IMMUNE NETWORK
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• v.20 no.4
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• pp.32.1-32.15
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• 2020

Influenza virus is the major cause of seasonal and pandemic flu. Currently, oseltamivir, a potent and selective inhibitor of neuraminidase of influenza A and B viruses, is the drug of choice for treating patients with influenza virus infection. However, recent emergence of oseltamivir-resistant influenza viruses has limited its efficacy. Morin hydrate (3,5,7,2',4'-pentahydroxyflavone) is a flavonoid isolated from Morus alba L. It has antioxidant, anti-inflammatory, neuroprotective, and anticancer effects partly by the inhibition of the NF-κB signaling pathway. However, its effects on influenza virus have not been studied. We evaluated the antiviral activity of morin hydrate against influenza A/Puerto Rico/8/1934 (A/PR/8; H1N1) and oseltamivir-resistant A/PR/8 influenza viruses in vitro. To determine its mode of action, we carried out time course experiments, and time of addition, hemolysis inhibition, and hemagglutination assays. The effects of the co-administration of morin hydrate and oseltamivir were assessed using the murine model of A/PR/8 infection. We found that morin hydrate reduced hemagglutination by A/PR/8 in vitro. It alleviated the symptoms of A/PR/8-infection, and reduced the levels of pro-inflammatory cytokines and chemokines, such as TNF-α and CCL2, in infected mice. Co-administration of morin hydrate and oseltamivir phosphate reduced the virus titers and attenuated pulmonary inflammation. Our results suggest that morin hydrate exhibits antiviral activity by inhibiting the entry of the virus.

• International Journal of Stem Cells
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• v.16 no.4
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• pp.425-437
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• 2023

Obesity, which continues to increase worldwide, was shown to irreversibly impair the differentiation potential and angiogenic properties of adipose tissue mesenchymal stromal cells (ADSCs). Because these cells are intended for regenerative medicine, especially for the treatment of inflammatory conditions, and the effects of obesity on the immunomodulatory properties of ADSCs are not yet clear, here we investigated how ADSCs isolated from former obese subjects (Ex-Ob) would influence macrophage differentiation and polarization, since these cells are the main instructors of inflammatory responses. Analysis of the subcutaneous adipose tissue (SAT) of overweight (OW) and Ex-Ob subjects showed the maintenance of approximately twice as many macrophages in Ex-Ob SAT, contained within the CD68⁺/FXIII-A^- inflammatory pool. Despite it, in vitro, coculture experiments revealed that Ex-Ob ADSCs instructed monocyte differentiation into a M2-like profile, and under inflammatory conditions induced by LPS treatment, inhibited HLA-DR upregulation by resting M0 macrophages, originated a similar percentage of TNF-α⁺ cells, and inhibited IL-10 secretion, similar to OW-ADSCs and BMSCs, which were used for comparison, as these are the main alternative cell types available for therapeutic purposes. Our results showed that Ex-Ob ADSCs mirrored OW-ADSCs in macrophage education, favoring the M2 immunophenotype and a mixed (M1/M2) secretory response. These results have translational potential, since they provide evidence that ADSCs from both Ex-Ob and OW subjects can be used in regenerative medicine in eligible therapies. Further in vivo studies will be fundamental to validate these observations.

• International Journal of Stem Cells
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• v.16 no.2
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• pp.191-201
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• 2023

Background and Objectives: O-cyclic phytosphingosine-1-phosphate (cP1P) is a synthetic chemical and has a structure like sphingosine-1-phosphate (S1P). S1P is known to promote cell migration, invasion, proliferation, and anti-apoptosis through hippocampal signals. However, S1P mediated cellular-, molecular mechanism is still remained in the lung. Acute lung injury (ALI) and its severe form acute respiratory distress syndrome (ARDS) are characterized by excessive immune response, increased vascular permeability, alveolar-peritoneal barrier collapse, and edema. In this study, we determined whether cP1P primed human dermal derived mesenchymal stem cells (hdMSCs) ameliorate lung injury and its therapeutic pathway in ALI mice. Methods and Results: cP1P treatment significantly stimulated MSC migration and invasion ability. In cytokine array, secretion of vascular-related factors was increased in cP1P primed hdMSCs (hdMSC^cP1P), and cP1P treatment induced inhibition of Lats while increased phosphorylation of Yap. We next determined whether hdMSC^cP1P reduce inflammatory response in LPS exposed mice. hdMSC^cP1P further decreased infiltration of macrophage and neutrophil, and release of TNF-α, IL-1β, and IL-6 were reduced rather than naïve hdMSC treatment. In addition, phosphorylation of STAT1 and expression of iNOS were significantly decreased in the lungs of MSC^cP1P treated mice. Conclusions: Taken together, these data suggest that cP1P treatment enhances hdMSC migration in regulation of Hippo signaling and MSC^cP1P provide a therapeutic potential for ALI/ARDS treatment.

• Journal of Microbiology and Biotechnology
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• v.34 no.6
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• pp.1287-1298
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• 2024

Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) that is currently difficult to treat effectively. Both Bacillus natto (BN) and ginseng-soluble dietary fiber (GSDF) are anti-inflammatory and helps sustain the intestinal barrier. In this study, the protective effects and mechanism of the combination of B. natto JLCC513 and ginseng-soluble dietary fiber (BG) in DSS-induced UC mice were investigated. Intervention with BG worked better than taking BN or GSDF separately, as evidenced by improved disease activity index, colon length, and colon injury and significantly reduced the levels of oxidative and inflammatory factors (LPS, ILs, and TNF-α) in UC mice. Further mechanistic study revealed that BG protected the intestinal barrier integrity by maintaining the tight junction proteins (Occludin and Claudin1) and inhibited the LPS/TLR4/NF-κB pathway in UC mice. In addition, BG increased the abundance of beneficial bacteria such as Bacteroides and Turicibacter and reduced the abundance of harmful bacteria such as Allobaculum in the gut microbiota of UC mice. BG also significantly upregulated genes related to linoleic acid metabolism in the gut microbiota. These BG-induced changes in the gut microbiota of mice with UC were significantly correlated with changes in pathological indices. In conclusion, this study demonstrated that BG exerts protective effect against UC by regulating the LPS/TLR4/NF-κB pathway and the structure and metabolic function of gut microbiota. Thus, BG can be potentially used in intestinal health foods to treat UC.