• Annals of Pediatric Endocrinology and Metabolism
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• 제23권4호
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• pp.204-209
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• 2018

Purpose: Growth hormone transduction defect (GHTD) is characterized by severe short stature, impaired STAT3 (signal transducer and activator of transcription-3) phosphorylation and overexpression of the cytokine inducible SH2 containing protein (CIS) and p21/CIP1/WAF1. To investigate the role of p21/CIP1/WAF1 in the negative regulation of the growth hormone (GH)/GH receptor and Epidermal Growth Factor (EGF)/EGF Receptor pathways in GHTD. Methods: Fibroblast cultures were developed from gingival biopsies of 1 GHTD patient and 1 control. The protein expression and the cellular localization of p21/CIP1/WAF1 was studied by Western immunoblotting and immunofluorescence, respectively: at the basal state and after induction with $200-{\mu}g/L$ human GH (hGH) (GH200), either with or without siRNA CIS (siCIS); at the basal state and after inductions with $200-{\mu}g/L$ hGH (GH200), $1,000-{\mu}g/L$ hGH (GH1000) or 50-ng/mL EGF. Results: After GH200/siCIS, the protein expression and nuclear localization of p21 were reduced in the patient. After successful induction of GH signaling (control, GH200; patient, GH1000), the protein expression and nuclear localization of p21 were reduced. After induction with EGF, p21 translocated to the cytoplasm in the control, whereas in the GHTD patient it remained located in the nucleus. Conclusion: In the GHTD fibroblasts, when CIS is reduced, either after siCIS or after a higher dose of hGH (GH1000), p21's antiproliferative effect (nuclear localization) is also reduced and GH signaling is activated. There also appears to be a positive relationship between the 2 inhibitors of GH signaling, CIS and p21. Finally, in GHTD, p21 seems to participate in the regulation of both the GH and EGF/EGFR pathways, depending upon its cellular location.

• Journal of Microbiology and Biotechnology
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• 제29권4호
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• pp.596-606
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• 2019

N-acyl-homoserine lactone quorum sensing (AHL-QS) has been shown to regulate many physiological behaviors in Serratia marcescens MG1. In the current study, the effects of AHL-QS on the biosynthesis of acid and neutral products by S. marcescens MG1 and its isogenic ${\Delta}swrI$ with or without supplementing exogenous N-hexanoyl-L-homoserine lactone ($C_6-HSL$) were systematically investigated. The results showed that swrI disruption resulted in rapid pH drops from 7.0 to 4.8, which could be restored to wild type by supplementing $C_6-HSL$. Furthermore, fermentation product analysis indicated that ${\Delta}swrI$ could lead to obvious accumulation for acidogenesis products such as lactic acid and succinic acid, especially excess acetic acid (2.27 g/l) produced at the early stage of fermentation, whereas solventogenesis products by ${\Delta}swrI$ appeared to noticeably decrease by an approximate 30% for acetoin during 32-48 h and by an approximate 20% for 2,3-butanediol during 24-40 h, when compared to those by wild type. Interestingly, the excess acetic acid produced could be removed in an AHL-QS-independent manner. Subsequently, quantitative real-time PCR was used to determine the mRNA expression levels of genes responsible for acidogenesis and solventogenesis and showed consistent results with those of product synthesis. Finally, by close examination of promoter regions of the analyzed genes, four putative luxI box-like motifs were found upstream of genes encoding acetyl-CoA synthase, lactate dehydrogenase, ${\alpha}$-acetolactate decarboxylase, and Lys-like regulator. The information from this study provides a novel insight into the roles played by AHL-QS in switching from acidogenesis to solventogenesis in S. marcescens MG1.

• 생명과학회지
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• 제28권12호
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• pp.1438-1447
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• 2018

본 연구는 추출 용매별 땅두릅 추출물과 Lactobacillus plantarum으로 발효한 후 용매별로 추출한 땅두릅 발효추출물을 이용하여 항염증 효과를 측정한 연구이다. 땅두릅은 중추신경 계통에 대한 흥분작용이 있고 혈압강하 작용이 알려져 있으며 관절염, 감기, 신경통, 류마티스, 피부가려움증 등에 쓰인다. LPS (lipopolysaccharide)로 염증을 유도한 마우스 유래 macrophage에서의 NO 생성 및 염증관련인자의 발현에 미치는 영향을 검토하여 항염증 소재로서의 이용 가능성을 확인하였다. 건조 땅두릅을 water, ethanol, hexane, ethyl acetate, butanol을 이용하여 추출한 추출물들과 L. plantarum으로 발효한 후 추출한 추출물들에서 RAW264.7 대식세포에 대한 독성 여부를 측정하였고, 세포에 대한 독성이 나타나지 않는 농도에서 추출물의 항염증 활성을 확인하였다. 본 실험에 사용된 추출물은 LPS로 유도된 NO 생성을 유의적으로 억제하였으며, 주요 염증 유발인자인 COX-2와 iNOS의 발현 또한 유의적으로 억제하는 효과를 나타냈다. 염증관련 cytokine인 $IL-1{\beta}$, IL-6 및 $TNF-{\alpha}$의 생성량 또한 유의적으로 감소시켰다. 이상의 결과로부터 땅두릅을 L. plantarum으로 발효한 후 water, ethanol, butanol로 추출하였을 때 염증 억제 효과가 있는 기능성 식품 소재로서의 개발 가능성이 있을 것으로 사료된다.

• 생명과학회지
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• 제29권10호
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• pp.1126-1135
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• 2019

세포 외로 단백질분해효소를 생산하는 효모 균주 CO-1을 대나무 부산물에서 분리하였다. CO-1은 원형 또는 타원형($3.1-4.0{\times}3.8-4.4{\mu}m$)으로, 생장을 위한 최적 온도는 $30^{\circ}C$, 초기 pH는 4.0이었다. 그리고 최대 15.0% (w/v)의 NaCl과 9.0%(v/v)의 ethanol 농도에서 생장하였다. 형태적, 생리 생화학적 특성 및 18S rRNA 유전자 염기서열을 통한 계통분석을 이용하여 동정을 실시한 결과 Pichia anomala로 판명되었다. P. anomala CO-1 단백질분해효소를 부분 정제한 결과 수율은 7.2％였으며, 정제 전에 비해 약 14.6배 정제되었다. Zymogram으로 측정한 효소의 분자량은 약 30 kDa으로 확인되었다. 본 균주는 배지 중에 탄소원과 질소원, 무기염으로 1.0%(w/v) CMC와 1.0%(w/v) yeast extract, 0.3%(w/v) $MnSO_4$를 사용하였을 경우 가장 높은 단백질분해효소 활성을 나타내었다. P. anomala CO-1이 생산하는 단백질분해효소의 최적 활성 pH와 온도는 각각 7.0과 $30^{\circ}C$였다. 또한 본 효소는 pH 4.0-10.0에서 75%의 안정성을 나타내었으며, $65^{\circ}C$에서 1시간 가열하여도 60％ 전후의 활성을 유지하였다. 균주의 효소 생산은 생육과 비례하였으며 대수증식기 후반에 최대의 효소 생산을 나타내었다.

• 한국자원식물학회지
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• 제32권2호
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• pp.153-159
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• 2019

이상의 연구 결과로 미루어 볼 때, 상동나무 가지 (STB)와 잎(STL)은 $GSK3{\beta}$와 ROS에 의존하지 않는 ${\beta}-catenin$의 분해 유도를 통해 대장암 및 폐암세포의 생육을 억제하는 것으로 나타났다. 본 결과는 상동나무 가지와 잎의 항암을 위한 대체보완소재 및 천연 항암제 개발을 위한 소재로 활용이 가능할 것으로 판단된다. 그러나 추가적 연구를 통해 상동나무 가지와 잎의 ${\beta}-catenin$의 분해 유도 관련 기전연구와 항암 활성물질의 분석연구가 필요할 것으로 사료된다.

• 한국자원식물학회지
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• 제32권2호
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• pp.109-115
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• 2019

이상의 연구 결과로 미루어 볼 때, 참식나무 기주 참나무겨우살이(TY-NS-B)는 $GSK3{\beta}$에 의한 ${\beta}$-catenin 인산화에 의존하지 않는 ${\beta}$-catenin의 분해 유도를 통해 암세포의 생육과 관련된 c-Myc의 발현을 억제하며, 이것은 비소세포성 폐암의 생육억제와 관련이 있는 것으로 판단된다. 또한, 본 결과는 항암을 위한 대체보완소재로 참식나무 기주 참나무겨우살이의 활용이 가능할 것으로 판단된다. 그러나 추가적 연구를 통해 참식나무 기주 참나무겨우살이가 어떻게 ${\beta}$-catenin의 분해 유도에 관여하는지에 대한 기전연구와 관련 활성물질의 분석연구가 필요할 것으로 사료된다.

• Journal of Nutrition and Health
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• 제54권5호
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• pp.435-447
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• 2021

Purpose: Body adiposity is negatively correlated with hepatic iron status, and Korean pine nut oil (PNO) has been reported to reduce adiposity. Therefore, we aimed to study the effects of PNO on adiposity, hepatic mineral status, and the expression of genes and proteins involved in iron absorption. Methods: Five-week-old male C57BL/6 mice were fed a control diet containing 10% kcal from PNO (PC) or soybean oil (SBO; SC), or a high-fat diet (HFD) containing 35% kcal from lard and 10% kcal from PNO (PHFD) or SBO (SHFD). Hepatic iron, copper, and zinc content; and expression of genes and proteins related to iron absorption were measured. Results: HFD-fed mice had a higher white fat mass (2-fold; p < 0.001), lower hepatic iron content (25% lower; p < 0.001), and lower hepatic Hamp (p = 0.028) and duodenal Dcytb mRNA levels (p = 0.037) compared to the control diet-fed mice. Hepatic iron status was negatively correlated with body weight (r = -0.607, p < 0.001) and white fat mass (r = -0.745, p < 0.001). Although the PHFD group gained less body weight (18% less; p < 0.05) and white fat mass (18% less; p < 0.05) than the SHFD group, the hepatic iron status impaired by the HFD feeding did not improve. The expression of hepatic and duodenal ferroportin protein was not affected by the fat amount or the oil type. PNO-fed mice had significantly lower Slc11a2 (p = 0.022) and Slc40a1 expression (p = 0.027) compared to SBO-fed mice. However, the PC group had a higher Heph expression than the SC group (p < 0.05). The hepatic copper and zinc content did not differ between the four diet groups, but hepatic copper content adjusted by body weight was significantly lower in the HFD-fed mice compared to the control diet-fed mice. Conclusion: HFD-induced obesity decreased hepatic iron storage by affecting the regulation of genes related to iron absorption; however, the 18% less white fat mass in the PHFD group was not enough to improve the iron status compared to the SHFD group. The hepatic copper and zinc status was not altered by the fat amount or the oil type.

• Animal Bioscience
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• 제34권2호
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• pp.243-255
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• 2021

Objective: Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that frequently contaminate maize and grain cereals, imposing risks to the health of both humans and animals and leading to economic losses. The gut microbiome has been shown to help combat the effects of such toxins, with certain microorganisms reported to contribute significantly to the detoxification process. Methods: We examined the cecum contents of three different dietary groups of pigs (control, as well as diets contaminated with 8 mg DON/kg feed or 0.8 mg ZEN/kg feed). Bacterial 16S rRNA gene amplicons were acquired from the cecum contents and evaluated by next-generation sequencing. Results: A total of 2,539,288 sequences were generated with ~500 nucleotide read lengths. Firmicutes, Bacteroidetes, and Proteobacteria were the dominant phyla, occupying more than 96% of all three groups. Lactobacillus, Bacteroides, Megasphaera, and Campylobacter showed potential as biomarkers for each group. Particularly, Lactobacillus and Bacteroides were more abundant in the DON and ZEN groups than in the control. Additionally, 52,414 operational taxonomic units were detected in the three groups; those of Bacteroides, Lactobacillus, Campylobacter, and Prevotella were most dominant and significantly varied between groups. Hence, contamination of feed by DON and ZEN affected the cecum microbiota, while Lactobacillus and Bacteroides were highly abundant and positively influenced the host physiology. Conclusion: Lactobacillus and Bacteroides play key roles in the process of detoxification and improving the immune response. We, therefore, believe that these results may be useful for determining whether disturbances in the intestinal microflora, such as the toxic effects of DON and ZEN, can be treated by modulating the intestinal bacterial flora.

• Journal of Nutrition and Health
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• 제53권6호
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• pp.570-582
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• 2020

본 연구는 상지추출물과 stilbene 화합물인 MSA, ORT의 항비만 효능을 비교 분석하고자 식이성 비만 마우스 모델을 이용하여 HFD에 실험물질을 첨가한 식이를 13주 동안 급여하였다. 연구결과, 실험물질들의 항비만 효능은 ORT, ME, MSA, MW군 순이었으며, 특히 ME와 ORT의 급여는 고지방식이에 의한 체중 및 체지방 증가, 혈당 증가, 이상지질혈증 및 지방간 개선에 있어 양성대조군인 GC군과 비슷한 효과를 보였다. 향후 관련 기작 규명 및 안전성 확보, 그리고 최대 유효성 도출을 위한 추출 조건 확립 및 용량 검증 등의 연구가 수행된다면 체중조절용 기능성 식품으로의 개발을 기대해 볼 수 있을 것으로 사료된다.

• 한국동물생명공학회지
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• 제35권4호
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• pp.297-306
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• 2020

The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.