• 한국식품과학회지
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• 제45권5호
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• pp.628-635
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• 2013

본 연구는 복분자 미숙과와 홍삼 물 추출물을 이용하여 인간 간암세포주(HepG2)와 위장관세포주(Caco-2)를 이용하여 콜레스테롤 억제효과 및 그와 연관된 HMG-CoA reductase 활성 억제효과 및 LDL 및 HDL과 관련된 분자기전을 조사하였다. 그 결과 복분자 미숙과와 홍삼 추출물은 모두 콜레스테롤 합성 억제 효과를 보였을 뿐만 아니라 HMG-CoA reductase 활성 억제효과를 보였다. 또한 두 추출물은 단독 투여시 보다 복합 투여시 부가효과를 보였다. 그리고 LDL receptor와 이를 조절하는 SREBP-2를 증가시키고 콜레스테롤 transport 인 ABCA1의 발현을 증가시켜 LDL를 낮추고 HDL를 높이는 효과가 있을 것으로 판단된다. 특히 두 추출물의 복합 투여시 훨씬 높은 시너지 효과를 나타냄을 확인하였다. 이러한 결과는 전통적으로 오랜 기간 사용되어온 천연물인 복분자와 홍삼이 콜레스테롤 예방에 효과적임을 입증한 결과로 관련 대사증후군의 예방에 기여할 것으로 사료된다.

• 생명과학회지
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• 제19권1호
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• pp.21-33
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• 2009

대황(Rhei Rhizoma; RR, 대황(大黃))은 Rheum officinale Baill.와 Rheum palmatum L.(polygonaceae)의 땅속부분으로 남아시아의 민속의학에서 간 및 신장의 손상을 치료하는데 널리 이용되고 있다. 본 연구에서는 배양한 흰쥐 해마신경세포의 저산소증모델을 이용하여 대황의 물추출물이 유전자 표현에 미치는 영향을 microarray 방법을 이용하여 조사하였다. 배양 후 10일 (DIV10)에 추출물을 배지에 $2.5{\mu}g/ml$ 농도로 첨가하고, DIV13에 저산소증(2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 h)을 유발한 후 24 시간 후에 total RNA를 분리하여 microarray에 사용하였다. MA-plot에 의하면 표현이 연화된 대부분의 유전자는 ${\pm}2$배 이내로 증감되었다. 이 가운데 Global M 값이 0.2(즉, 15%)보다 더 증가한 유전자는 472종, Global M 값이 -0.2(즉, -15%)보다 더 감소한 유전자는 725종이였다. 세포의 생존과 관련된 유전자 가운데 세포자연사 억제유전자인 Tegt(2.4배), Nfkb1 (2.4배), Veg (1.8배), Ngfr (1.6배) 등이 크게 증가하였으며, 반면에 자연사 촉진유전자인 Bad (-64%), Cstb (-66%)는 감소하였다. 스트레스를 극복하는데 필요한 유전자인 Defb3 (2.7배), Cygb (2.2배), Ahsg (2.18배), Alox5 (2배) 등도 크게 증가하였다. 그리고 세포 성장을 촉진하는 유전자인 Erbb2 (1.84배), Mapk12 (1.8배)도 크게 증가하였다. 따라서 대황의 물추출물은 세포생존에 필요한 유전자를 증가시키고, 세포사를 유도하는 유전자는 감소시킴으로서 저산소층 스트레스에서 신경세포의 사망을 억제하는 것으로 해석된다.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.71-77
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• 2010

The time, yield, and related genes expression of PHB accumulation of Acidiphilium cryptum DX1-1 were investigated under four different initial C/N ratios, 1.2, 2.4, 7.5, and 24. The results of time and yield of poly-$\beta$-hydroxybutyrate (PHB) accumulation show that the initial C/N ratio of 2.4 was optimum for strain DX1-1 to accumulate PHB, but both higher and lower initial C/N ratios did not favor that process. Based on the genome of Acidiphilium cryptum JF-5, 13 PHB accumulation related genes in strain JF-5 were chosen and successfully cloned from strain DX1-1. The differential expressions of the 13 functional genes, in different C/N ratios as cited above, were then studied by real-time PCR. The results show that all the 13 genes were most upregulated when the initial C/N ratio was 2.4, and among which the gene Acry_3030 encoding poly-$\beta$-hydroxybutyrate polymerase and Aery_0626 encoding acetyl-CoA synthetase were much more upregulated than the other genes, which proved that they play the most important role for PHB accumulation, and acetate is the main initial substance for PHB accumulation for strain DX1-1. Potential regulatory motifs analysis showed that the genes related to PHB accumulation are regulated by different promoters and that the motif had weak similarity to the model promoters, suggesting that PHB metabolism in Acidiphilium cryptum may be mediated by a different mechanism.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4607-4611
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• 2012

Objective: Endometrial cancer (EC) is the most common gynecologic malignancy. Identification of potential biomarkers of EC would be helpful for the detection and monitoring of malignancy, improving clinical outcomes. Methods: The Weighted Gene Co-expression Network Analysis method was used to identify prognostic markers for EC in this study. Moreover, underlying molecular mechanisms were characterized by KEGG pathway enrichment and transcriptional regulation analyses. Results: Seven gene co-expression modules were obtained, but only the turquoise module was positively related with EC stage. Among the genes in the turquoise module, COL5A2 (collagen, type V, alpha 2) could be regulated by PBX (pre-B-cell leukemia homeobox 1)1/2 and HOXB1(homeobox B1) transcription factors to be involved in the focal adhesion pathway; CENP-E (centromere protein E, 312kDa) by E2F4 (E2F transcription factor 4, p107/p130-binding); MYCN (v-myc myelocytomatosis viral related oncogene, neuroblastoma derived [avian]) by PAX5 (paired box 5); and BCL-2 (B-cell CLL/lymphoma 2) and IGFBP-6 (insulin-like growth factor binding protein 6) by GLI1. They were predicted to be associated with EC progression via Hedgehog signaling and other cancer related-pathways. Conclusions: These data on transcriptional regulation may provide a better understanding of molecular mechanisms and clues to potential therapeutic targets in the treatment of EC.

• Asian-Australasian Journal of Animal Sciences
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• 제31권8호
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• pp.1345-1357
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• 2018

Objective: This study examined the influence of dietary supplementation of Nigella sativa seeds, Rosmarinus officinalis leaves and their combination on carcass attributes, fatty acid (FA) composition, gene expression, lipid oxidation and physicochemical properties of longissimus dorsi (LD), semitendinosus (ST), and supraspinatus (SS) muscles in Dorper lambs. Methods: Twenty-four Dorper lambs ($18.68{\pm}0.6kg$, 4 to 5 months old) were randomly assigned to a concentrate mixture containing either, no supplement (control, T1), 1% Rosmarinus officinalis leaves (T2), 1% Nigella sativa seeds (T3), or 1% Rosmarinus officinalis leaves+1% Nigella sativa seeds (T4) on a dry matter basis. The lambs were fed the treatments with urea-treated rice straw for 90 days, slaughtered and the muscles were subjected to a 7 d postmortem chill storage. Results: The T2 lambs had greater (p<0.05) slaughter and cold carcass weights than the control lambs. Dietary supplements did not affect (p>0.05) chill loss, dressing percentage, carcass composition, intramuscular fat and muscle pH in Dorper lambs. Meat from supplemented lambs had lower (p<0.05) cooking and drip losses, shear force, lightness, and lipid oxidation and greater (p<0.05) redness compared with the control meat. The impact of dietary supplements on muscle FA varied with muscle type. Diet had no effect (p>0.05) on the expression of stearoyl-CoA desaturase and lipoprotein lipase genes in LD and ST muscles in Dorper lambs. The T2 and T3 diets up regulated the expression of AMP-activated protein kinase alpha 2 gene in LD and ST muscles and up regulated the expression of sterol regulatory element-binding protein 1 in ST muscle in Dorper lambs. Conclusion: Dietary supplementation of Nigella sativa seeds and Rosmarinus officinalis leaves had beneficial effects on meat quality in Dorper lambs.

• Biomolecules & Therapeutics
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• 제20권3호
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• pp.299-305
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• 2012

Endochondral bone formation is the process by which mesenchymal cells condense to become chondrocytes, which ultimately form new bone. The process of chondrogenic differentiation and hypertrophy is critical for bone formation and as such is regulated by many factors. In this study, we aimed to indentify novel factors that regulate chondrogenesis. We investigated the possible role of isopsoralen in induction of chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Isopsoralen treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. Further, ATDC5 cells treated with isopsoralen were stained more intensely with Alcian blue than control cells, suggesting that isopsoralen increases the synthesis of matrix proteoglycans. Similarly, isopsoralen markedly induced the activation of alkaline phosphatase activity compared with control cells. Isopsoralen enhanced the expressions of chondrogenic marker genes such as collagen II, collagen X, OCN, Smad4 and Sox9 in a time-dependent manner. Furthermore, isopsoralen induced the activation of extracellular signal-regulated kinase (ERK) and p38 MAP kinase, but not that of c-jun N-terminal kinase (JNK). Isopsoralen significantly enhanced the protein expression of BMP-2 in a time-dependent manner. PD98059 and SB 203580, inhibitors of ERK and p38 MAPK, respectively, decreased the number of stained cells treated with isopsoralen. Taken together, these results suggest that isopsoralen mediates a chondromodulating effect by BMP-2 or MAPK signaling pathways, and is therefore a possible therapeutic agent for bone growth disorders.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2014년도 추계학술대회 및 정기총회
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• pp.13-13
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• 2014

Species belonging to the genus Fusarium are widely distributed and cause diseases in many plants. Isolation of fungal strains from air or cereals is necessary for disease forecasting, disease diagnosis, and population genetics [1]. Previously we showed that Fusarium species are resistant to toxoflavin produced by the bacterial rice pathogen Burkholderia glumae while other fungal genera are sensitive to the toxin, resulting in the development of a selective medium for Fusarium species using toxoflavin [2]. In this study, we have tried to elucidate the resistant mechanism of F. graminearum against toxoflavin and interaction between the two pathogens in nature. To test whether B. glumae affects the development of F. graminearum, the wild-type F. graminearum strains were incubated with either the bacterial strain or supernatant of the bacterial culture. Both conditions increased the conidial production five times more than when the fungus was incubated alone. While co-incubation resulted in dramatic increase of conidial production, conidia germination delayed by either the bacterial strain or supernatant. These results suggest that certain factors produced by B. glumae induce conidial production and delay conidial germination in F. graminearum. To identify genes related to toxoflavin resistance in F. graminearum, we screened the transcriptional factor mutant library previously generated in F. graminearum [3] and identified one mutant that is sensitive to toxoflavin. We analyzed transcriptomes of the wild-type strain and the mutant strain under either absence or presence of toxoflavin through RNAseq. Expression level of total genes of 13,820 was measured by reads per kilobase per million mapped reads (RPKM). Under the criteria with more than two-fold changes, 1,440 genes were upregulated and 1,267 genes were down-regulated in wild-type strain than mutant strain in response to toxoflavin treatment. A comparison of gene expression profiling between the wild type and mutant through gene ontology analysis showed that genes related to metabolic process and oxidation-reduction process were highly enriched in the mutant strain. The data analyses will focus on elucidating the resistance mechanism of F. graminearum against toxoflavin and the interaction between the two pathogens in rice. Further evolutionary history will be traced through figuring out the gene function in populations and in other filamentous fungi.

• 원예과학기술지
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• 제29권6호
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• pp.623-632
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• 2011

본 연구는 배추에서 항암물질 PEITC의 함량을 높이기 위하여 PEITC 대사과정에서 관련 유전자인 myrosinase (MYR)와 Glutathione S-transferase(GST) 유전자를 분리하고 Agrobacterium tumefacien 형질전환 방법을 통하여 유전자 발현을 조절하였다. 분리된 MYR과 GST의 cDNA는 각각 1647bp와 624bp임을 확인하였고 pET system으로 단백질의 발현을 확인하였다. 형질전환을 위해서 MYR-과발현 벡터와 GST-발현억제 벡터를 제작하였으며 이를 이용하여 배추에 형질전환한 후 PCR 검정을 통해 MYR-과발현 벡터로 형질전환된 개체(IMS) 13개체를 GST-발현억제 벡터로 형질전환된 개체(IGA) 5개체를 선발하였다. 선발된 $T_0$ 개체는 $T_1$ 세대로 진전시켰으며 $T_1$ 형질전환 계통의 서던분석 결과 배추 genome내로 1-4 copy의 T-DNA가 삽입된 것을 확인하였다. 유전자 발현양을 real-time RT PCR로 조사한 결과 IMS는 발현량이 1.03-4.25배 증가하였고 IGA는 26.42-42.22배 감소하였다. IMS와 IGA의 각 계통에서 PEITC의 농도를 GC-MS 방법을 이용하여 확인한 결과 IMS는 PEITC 함량이 형질전환이 되지 않은 대조군에 비해 최대 4.86배까지 증가한 계통을 확인하였고 IGA는 최대 3.89배까지 증가된 계통을 확인하였다. 최종적으로 본 연구를 통하여 항암물질 PEITC량의 증가를 보인 형질전환계통 IMS 1, 3, 5, 12, 15 및 IGA 1, 2, 4를 선발하였다.

• Animal Bioscience
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• 제34권5호
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• pp.801-810
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• 2021

Objective: microRNAs (miRNAs) can play a role in a variety of physiological and pathological processes, and their role is achieved by regulating the expression of target genes. Our previous high-throughput sequencing found that ssc-miR-185 plays an important regulatory role in piglet diarrhea, but its specific target genes and functions in intestinal porcine epithelial cell (IPEC-J2) are still unclear. We intended to verify the target relationship between porcine miR-185 and cell division cycle 42 (CDC42) gene in IPEC-J2 and to explore the effect of miR-185 on the proliferation of IPEC-J2 cells. Methods: The TargetScan, miRDB, and miRanda software were used to predict the target genes of porcine miR-185, and CDC42 was selected as a candidate target gene. The CDC42-3' UTR-wild type (WT) and CDC42-3'UTR-mutant type (MUT) segments were successfully cloned into pmirGLO luciferase vector, and the luciferase activity was detected after co-transfection with miR-185 mimics and pmirGLO-CDC42-3'UTR. The expression level of CDC42 was analyzed using quantitative polymerase chain reaction and Western blot. The proliferation of IPEC-J2 was detected using cell counting kit-8 (CCK-8), methylthiazolyldiphenyl-tetrazolium bromide (MTT), and 5-ethynyl-2'-deoxyuridine (EdU) assays. Results: Double enzyme digestion and sequencing confirmed that CDC42-3'UTR-WT and CDC42-3'UTR-MUT were successfully cloned into pmirGLO luciferase reporter vector, and the luciferase activity was significantly reduced after co-transfection with miR-185 mimics and CDC42-3'UTR-WT. Further we found that the mRNA and protein expression level of CDC42 were down-regulated after transfection with miR-185 mimics, while the opposite trend was observed after transfection with miR-185 inhibitor (p<0.01). In addition, the CCK-8, MTT, and EdU results demonstrated that miR-185 promotes IPEC-J2 cells proliferation by targeting CDC42. Conclusion: These findings indicate that porcine miR-185 can directly target CDC42 and promote the proliferation of IPEC-J2 cells. However, the detailed regulatory mechanism of miR-185/CDC42 axis in piglets' resistance to diarrhea is yet to be elucidated in further investigation.

• IMMUNE NETWORK
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• 제1권1호
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• pp.14-25
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• 2001

Background: Bone marrow stromal cells (BMSCs) express many cell surface molecules, which regulate the proliferation and differentiation of immune cells within the bone marrow. Methods: To identify cell surface molecules, which can regulate cell proliferation through cell interaction, monoclonal antibodies (MoAbs) against BMSCs were produced. Among them, 1H8 MoAb, which recognized distinctly an 80 kDa protein, abolished myeloma cell proliferation that was induced by co-culturing with BMSCs. Results: IL-6 gene expression was increased when myeloma or stromal cells were treated with 1H8 MoAb. In addition, the expression of IL-6 receptor and CD40 was up-regulated by 1H8 treatment, suggesting that the molecule recognized by 1H8 MoAb is involved in cell proliferation by modulating the expression of cell growth-related genes. Myeloma cells contain high levels of reactive oxygen species (ROS), which are related to gene expression and tumorigenesis. Treatment with 1H8 decreased the intracellular ROS level and increased PAG antioxidant gene concomitantly. Finally, 1H8 induced the tyrosine phosphorylation of several proteins in U266. Conclusion: Taken together, 1H8 MoAb recognized the cell surface molecule and triggered the intracellular signals, which led to modulate gene expression and cell proliferation.