• 생명과학회지
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• 제28권6호
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• pp.639-647
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• 2018

한국의 고급 양식대상 어종인 붉바리(Epinephelus akaara)로부터 새로운 heat shock protein (Hsp) 70을 동정하였다. 붉바리 Hsp70 (RgHsp70)의 cDNA는 RACE (rapid amplification of cDNA ends)법을 사용하였고, RgHsp70 cDNA의 전장은 2,152 bp이고, 5'-terminal untranslated region (UTR)은 105 bp, 3'-terminal UTR은 274 bp, 590개의 아미노산을 암호화하는 open reading frame (ORF)는 1,773 bp였으며, 분자무게(molecular weight)는 64.9 kDa 및 등전위값(isoelectric point, pI)은 5.2였다. 추정되는 아미노산 비교 및 계통발생학적 분석 결과, 다른 어종과 마찬가지로 Hsp70 고유의 signature를 포함하는 것을 비롯하여 높은 유사성을 나타내었으므로 RgHsp70이 Hsp70 family임을 확인할 수 있었다 RgHsp70 mRNA는 간과 두신 조직에서 높은 발현을 보였으며, 48시간 동안 수온별(21, 18, 15 및 $12^{\circ}C$) 노출 후 간 조직에서 대조구인 $21^{\circ}C$보다 $12^{\circ}C$에서 발현이 증가함을 확인하였다. 본 연구에서는, 수온이 하강함에 따라 RgHsp70 mRNA 발현에 주요한 영향을 미치는 것으로 보아, 수온변화에 따른 스트레스로 인해 발현의 변화를 나타내는 주요 스트레스성 단백질임을 확인할 수 있었다.

• Molecules and Cells
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• 제25권1호
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• pp.55-63
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• 2008

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.

• Journal of Animal Science and Technology
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• 제62권4호
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• pp.504-520
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• 2020

Proliferation of shrubs at the expense of native forage in pastures has been associated with large changes in dry-matter intake and dietary components for grazing ruminants. These changes can also affect the animals' physiology and metabolism. However, little information is available concerning the effect of pastoral-shrub grazing on the rumen bacterial community. To explore rumen bacteria composition in grazing yaks and the response of rumen bacteria to increasing shrub coverage in alpine meadows, 48 yak steers were randomly assigned to four pastures with shrub coverage of 0%, 5.4%, 11.3%, and 20.1% (referred as control, low, middle, and high, respectively), and ruminal fluid was collected from four yaks from each pasture group after 85 days. Rumen fermentation products were measured and microbiota composition determined using Ion S5™ XL sequencing of the 16S rRNA gene. Principal coordinates analysis (PCoA) and similarity analysis indicated that the degree of shrub coverage correlated with altered rumen bacterial composition of yaks grazing in alpine shrub meadows. At the phyla level, the relative abundance of Firmicutes in rumen increased with increasing shrub coverage, whereas the proportions of Bacteroidetes, Cyanobacteria and Verrucomicrobia decreased. Yaks grazing in the high shrub-coverage pasture had decreased species of the genus Prevotellaceae UCG-001, Lachnospiraceae XPB1014 group, Lachnospiraceae AC2044 group, Lachnospiraceae FCS020 group and Fretibacterium, but increased species of Christensenellaceae R-7 group, Ruminococcaceae NK4A214 group, Ruminococcus 1, Ruminococcaceae UCG-002, Ruminococcaceae UCG-005 and Lachnospiraceae UCG-008. These variations can enhance the animals' utilization efficiencies of cellulose and hemicellulose from native forage. Meanwhile, yaks grazed in the high shrub-coverage pasture had increased concentrations of ammonia nitrogen (NH₃-N) and branched-chain volatile fatty acids (isobutyrate and isovalerate) in rumen compared with yaks grazing in the pasture without shrubs. These results indicate that yaks grazing in a high shrub-coverage pasture may have improved dietary energy utilization and enhanced resistance to cold stress during the winter. Our findings provide evidence for the influence of shrub coverage on the rumen bacterial community of yaks grazing in alpine meadows as well as insights into the sustainable production of grazing yaks on lands with increasing shrub coverage on the Qinghai-Tibet Plateau.

• 한국식품과학회지
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• 제37권3호
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• pp.456-464
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• 2005

본 연구에서는 최근 전세계적으로 문제가 되고 있는 항생제 내성균에 대해 그 항균효과를 천연의 약용식물을 이용하여 분자생물학적 수준에서 비교해보고 천연항균제의 개발에 우선하여 직접 식품에 첨가하여 그 효과를 검색하고자 하였다. 병원내감염의 주된 원인균으로 알려진 황색포도상구균(Staphylococcusaureus)은 여러 항균제에 대하여 내성을 획득함으로써 질병의 치료를 어렵게 한다는 점에서 매우 심각한 문제이다. 이러한 내성균은 다양한 방법으로 동 식물에 빠르게 전파되고 있으므로 소비자가 내성균을 지닌 식품을 섭취해 잠재적인 내성균을 보유하게 되는 결과를 낳게 된다. 이러한 항생제 내성 황색포도상구균(Methicillin Resistant S. aureus, MRSA)에 대해 천연 약용 식물 중 우수한 항균력을 가지는 감초를 순차 분획하여 각 분획물을 이용한 MRSA에 대한 항균 활성을 알아본 결과 hexane, chloroform 분획물에서 항균활성을 나타냈으며, 동일농도의 생육저해비교 실험에서 대부분의 내성균주에 대해 대조군과 유사한 효과를 나타낸 penicillin에 비해 감초분획물이 대수기를 넘는 억제효과를 나타내었으며 이 중 chloroform 분획물이 가장 높은 저해활성을 나타내었다. 또한 RT-PCR을 통하여 표준균주와 내성균주의 내성 정도와 내성 유전자를 관찰해본 결과 KCCM 40510은 내성획득유전자인 mecA, mecRI, mecI, femA에 발현이 높게 나타나 고도내성균임을 알 수 있었으며, 고도내성을 나타내는 균주를 이용하여 감초의 chloroform 분획물을 농도별로($50,\;100,\;250\;{\mu}/mL$) 처리한 결과 농도 의존적으로 유전자 발현이 억제됨을 볼 수 있었다. 특히 고도내성균주인 KCCM 40510은 메티실린 내성을 상승시키는 mecA 유전자의 발현을 억제시키는 mecRI 유전자의 pathway와 상응하는 것으로 보여지므로 감초가 이러한 고도내성균에 효과가 있음을 알 수 있었다. 또한 SEM을 통한 형태학적 관찰을 통해 분획물 처리 시 세포벽이 파괴되는 것을 확인할 수 있었다. 한편 이러한 감초추출물을 직접 식품에 첨가하여 내성 균주에 대한 억제 효과도 확인한 결과 추출물 200mg/100mL 농도에서 포도상구균에 대해 항균효과를 나타내는 것을 볼 수 있었다.

• 생명과학회지
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• 제17권9호통권89호
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• pp.1204-1210
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• 2007

Totopoisomerase II 저해제인 etoposide는 핵안에 DNA double strand breaks를 일으키므로써 세포의 DNA에 손상을 초래한다. 본 연구에서는 인간 망막색소상피세포인 ARPE-19 세포에서의 세포성장 및 아폽토시스에서 etoposide의 역할을 살펴보았다. Etoposide는 세포의 성장을 크게 감소시켰으며 TUNEL에서 아폽토시스를 나타내는 DNA fragmentation의 증가를 유도하였다. 게다가, etoposide는 산화적 손상에 대해 세포나 조직을 보호하는 역할을 하는 것으로 알려진 세포내 항산화효소인 heme oxygenase-1 (HO-1)의 발현을 크게 증가시켰다. Etoposide에 의한 HO-1 발현증가는 항산화물질 NAC에 의해 억제되었는데, 이는 etoposide에 의한 세포내 ROS의 증가가 HO-1 발현에 중요한 역할을 한다는 것을 의미한다. 또한 HO-1 발현을 억제하기 위하여 HO-1 siRNA 방법을 사용하였다. 흥미롭게도, HO-1 유전자의 knock-down은 etoposide에 의해 유도되는 DNA fragmentation의 정도를 증가시켰다. 이들 결과를 종합해볼 때, etoposide에 의해 자극되어진 ARPE-19 세포에서 발현증가된 HO-1은 etoposide에 의한 아폽토시스 유발과정에서 세포를 보호하는 항아폽토시스의 기능을 한다는 것을 시사한다.

• Bulletin of the Korean Chemical Society
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• 제32권6호
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• pp.2051-2057
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• 2011

Several studies have demonstrated that nanoparticles (NPs) have toxic effects on cultured cell lines, yet there are no clear data describing the overall molecular changes induced by NPs currently in use for human applications. In this study, the in vitro cytotoxicity of three oxide NPs of around 100 nm size, namely, mesoporous silica (MCM-41), iron oxide ($Fe_2O_3$-NPs), and zinc oxide (ZnO-NPs), was evaluated in the human embryonic kidney cell line HEK293. Cell viability assays demonstrated that 100 ${\mu}g/mL$ MCM-41, 100 ${\mu}g/mL$ $Fe_2O_3$, and 12.5 ${\mu}g/mL$ ZnO exhibited 20% reductions in HEK293 cell viability in 24 hrs. DNA microarray analysis was performed on cells treated with these oxide NPs and further validated by real time PCR to understand cytotoxic changes occurring at the molecular level. Microarray analysis of NP-treated cells identified a number of up- and down-regulated genes that were found to be associated with inflammation, stress, and the cell death and defense response. At both the cellular and molecular levels, the toxicity was observed in the following order: ZnO-NPs > $Fe_2O_3$-NPs > MCM-41. In conclusion, our study provides important information regarding the toxicity of these three commonly used oxide NPs, which should be useful in future biomedical applications of these nanoparticles.

• 생명과학회지
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• 제21권5호
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• pp.664-670
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• 2011

Heat shock protein (HSP)은 외부적인 자극에 반응하여 세포를 보호하는 역할을 한다. 또한 HSP90은 혈관질환에서 처럼 세포가 사멸되거나 손상을 입는 경우 세포 밖으로 유리된다. 그러나 지금까지 세포 밖 HSP90이 혈관평활근세포에 어떠한 영향을 주는지에 대한 연구는 미약하다. 따라서 본 연구는 혈관평활근세포에서 HSP90이 CXCL10 발현에 대한 영향과 그 기전을 규명하였다. HSP90에 노출된 혈관평활근세포에서 CXCL10 transcript가 증가하고, CXCL10 단백질의 분비가 증가되었다. HSP90에 의한 CXCL10 분비는 Toll-like receptor (TLR)-2/-4 억제제인 1-palmitoyl-2-arachidonosyl-sn-phosphatidylcholine (OxPAPC)과 NADPH oxidase 억제제인 diphenyleneiodium 그리고 Akt 경로를 억제하는 LY294002와 Akti IV에 의하여 크게 감소되었다. 또한 TLR-4의 dimerization을 저해하는 curcumin 역시 HSP90에 의한 CXCL10의 분비를 억제하였다. 전사인자인 nuclear factor kappa B(NF-${\kappa}$B)의 생물학적 억제제인 inhibitory kappa B (I${\kappa}$B)와 NF-${\kappa}$B 억제작용이 있는 rasveratrol은 HSP90에 의한 CXCL10 분비를 억제하였다. 이러한 결과는 혈관평활근세포에서 HSP90에 의한 CXCL10의 발현에 TLR-4와 Akt 및 NF-${\kappa}$B가 관여함을 의미한다.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.13-34
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• 2003

Fructose-1, 6-diphosphate (FOP), a glycolytic metabolite is reported to ameliorate inflammation and inhibit the nitric oxide production in murine macrophages stimulated with endotoxin. It is also reported that FOP has cytoprotective effects against hypoxia or ischemia/reperfusion injury in brain and heart. In this study, we examined whether FDP has protective effects on UV-induced oxidative damage in skin cell culture system and human skin in vivo. FDP had a protective role in UVB-induced LDH release and ROS accumulation in HaCaT although it did not show direct radical scavenging effect in the experiment using 1, 1-diphenyl-2-picrylhydrazyl (DPPH). FDP also preserved cellular GSH content after UV irradiation in HaCaT and normal human fibroblast culture system. Cellular oxidative stress induces multiple downstream signaling pathways that regulate expression of multiple gene including MMP-1 and collagen, we examined the effects of FDP on UV-induced alteration of these protein expression in fibroblast culture and human skin in vivo. The increased MMP-1 expression in fibroblast and human skin by UV irradiation was significantly decreased by FDP. FDP also prevented the UV-induced decrease of collagen expression in fibroblast and human skin. Moreover, the decreasing the intracellular levels of reducing equivalents in human fibroblast by glutathione (GSH) depletion lowered the UVA dose threshold for reduction of procollagen expression, indicating that the differences of glutathione contents define the susceptibility of fibroblasts towards UV-induced reduction of procollagen expression. FDP also preserved cellular GSH content after UV irradiation, indicating that FDP has protective effects on UV-induced reduction of procollagen expression, which are possibly through maintaining intracellular reducing equivalent. Based on these premises, we examined the effect of daily use of a moisturizer containing FDP on facial wrinkle in comparison with vehicle moisturizer lacking FDP. In the clinical study, FDP significantly decreased facial wrinkle compared with vehicle alone after 6 months of use. Our results suggest that FDP has anti-aging effects in skin by increasing cellular antioxidant system and preventing oxidative signal and inflammatory reaction. Therefore FDP may be useful anti-aging agent for cosmetic purpose.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2707-2712
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• 2015

Background: The human homologue of the mouse double minute 2 (MDM2) gene is a negative regulator of Tp53. MDM2-309T>G a functional promoter polymorphism was found to be associated with overexpression thereby attenuation of Tp53 stress response and increased cancer susceptibility. We have planned to evaluate the possible role of MDM2-309T>G polymorphism with risk and response to chemotherapy in AML. Materials and Methods: A total of 223 de novo AML cases and 304 age and sex matched healthy controls were genotyped for the MDM2-309T>G polymorphism through the tetra-primer amplification refractory mutation system (ARMS)-PCR method. In order to assess the functional relationship of -309T>G SNP with MDM2 expression level, we quantified MDM2 mRNA in 30 primary AML blood samples through quantitative RT-PCR. Both the (-309T>G) genotypes and the MDM2 expression were correlated with disease free survival (DFS) rates among patients who have achieved complete remission (CR) after first induction chemotherapy. Results: MDM2-309T>G polymorphism was significantly associated with AML development (p<0.0001). The presence of either GG genotype or G allele at MDM2-309 confered 1.79 (95% CI: 1.12-2.86; p<0.001) and 1.46 fold (95%CI: 1.14-1.86; p= 0.003) increased AML risk. Survival analysis revealed that CR+ve cases with GG genotype had significantly increased DFS rates (16months, p=0.05) compared to CR+ve TT (11 months) and TG (9 months) genotype groups. Further, MDM2 expression was also found to be significantly elevated in GG genotype patients (p=0.0039) and among CR+ve cases (p=0.0036). Conclusions: The MDM2-309T>G polymorphism might be involved in AML development and also serve as a good prognostic indicator.

• 한국환경과학회지
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• 제25권6호
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• pp.789-798
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• 2016

Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.