• Asian-Australasian Journal of Animal Sciences
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• 제27권5호
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• pp.749-756
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• 2014

The present study was carried out to investigate the effects of dietary antioxidants on pro-inflammatory cytokines, heat shock protein (HSP) and antioxidant status in broiler chicks under summer conditions. A total of 162, 3-d-old broiler chicks were randomly assigned to a basal diet (CON) and the basal diet supplemented with vitamin C (200 mg/kg diet, VCD) or vitamin E (100 mg/kg, VED) until 35 day of age. All birds were exposed to summer diurnal heat stress at average daily fluctuations of temperature between $32^{\circ}C$ to $34^{\circ}C$ at day to $27^{\circ}C$ to $29^{\circ}C$ at night for the entire feeding periods. There was no significant difference in body weight, feed to gain ratio and the relative organ weight except the thymus in response to dietary vitamin C or E supplementation. However, the mRNA expression of interleukin (IL)-$1{\beta}$, IL-6, interferon (IFN)-${\gamma}$, Toll like receptor (TLR)-4 and HSP70 in the liver of birds fed diet containing vitamin C significantly (p<0.05) decreased compared with those in birds fed basal diet. Dietary vitamin E also showed a significant (p<0.05) decrease in the mRNA expression of IL-6 and HSP70 compared with a basal diet. Total antioxidant status (TAS) in serum of birds fed vitamin C supplemented diet was significantly (p<0.05) higher with than that in birds a basal diet. Lipid peroxidation in serum and liver resulted in a significant (p<0.05) decrease in response to dietary vitamin C or E supplementation. In conclusion, dietary supplementation with antioxidant vitamins, especially vitamin C resulted in a significant decrease in the mRNA expression of pro-inflammatory cytokines and HSP70, and higher antioxidant parameters than that of birds on the basal diet under summer conditions.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.946-951
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• 2006

To study the effect of iron on Saccharomyces cerevisiae, whole-cell proteins of Saccharomyces cerevisiae were extracted and subjected to two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and differentially expressed proteins were identified. The proteins separated were further identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and were compared with a protein database. Of more than 300 spots separated by molecular weight and isoelectric points, 27 differentially expressed spots were identified. Ten proteins were found to be differentially expressed at high iron concentration. Triosephosphate isomerase (TPI), YDR533C hypothetical protein, superoxide dismutase (SOD), 60 kDa heat-shock protein (HSP60), pyruvate dehydrogenase beta subunit 1 (PDB1), and old yellow enzyme 2 (OYE2) were upregulated, whereas thiol-specific antioxidant (TSA), regulatory particle non-ATPase subunit 8 (RPN8), thiol-specific peroxiredoxin 1 (AHP1), and fructose-1, 6-bisphosphate adolase (FBA) were downregulated by iron. Based on the result, we propose that SOD upregulated by iron would protect the yeast from oxidative stress by iron, and that TSA downregulated by iron would render cells hypersensitive to oxidative stress.

• Journal of Microbiology and Biotechnology
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• 제20권3호
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• pp.542-549
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• 2010

In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin $\beta$-subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at $90^{\circ}C$ for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}$ and $50^{\circ}$, respectively. Specifically, the activity of malate dehydrogenase (MDH) at $85^{\circ}$ was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.

• The Korean Journal of Physiology and Pharmacology
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• 제14권4호
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• pp.205-212
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• 2010

Endometritis is one of the primary reasons for reproductive failure. In order to investigate endometritis-associated marker proteins, proteomic analysis was performed on bovine endometrium with endometritis. In bovine endometritis, desmin, $\alpha$-actin-2, heat-shock protein (HSP) 27, peroxiredoxin-6, luteinizing hormone receptor isoform 1, collectin-43 precursor, deoxyribonuclease-I (DNase-I), and MHC class I heavy chain (MHC-Ih) were up-regulated. In contrast, transferrin, interleukin-2 precursor, hemoglobin $\beta$ subunit, and potassium channel tetramerisation domaincontaining 11 (KCTD11) were down-regulated in comparison to normal endometrium. The proteomic results were validated by semiquantitative-PCR and immunoblot analysis. The mRNA levels of desmin, transferrin, $\alpha$-actin-2, HSP27, KCTD11, and MHC-Ih were up-regulated by over 1.5-fold, and showed a pattern similar to their proteomic profiles. Desmin and $\alpha$-actin-2 protein showed positive correlations between proteomic analysis and immunoblot analysis. These results suggest that desmin and $\alpha$-actin-2 may play important roles in endometritis-related function, and could be useful markers for the diagnosis of bovine endometritis.

• 대한본초학회지
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• 제20권4호
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• pp.1-10
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• 2005

Objectives : We investigated the anti-aging effect on skin with the extract of the needles of red pine, Pinus densiflora. Methods : We measured various effects related to skin such as the anti-oxidant effect, the protection against ultraviolet (UV) irradiation, the inhibition of reactive oxygen species (ROS) generation, the induction of heat shock protein 70 (HSP70), the reduction of matrix metalloproteinase-2 (MMP-2) synthesis and senescent cell. Results : The results were as follows : The extract of the needles of red pine (RP) had the potent anti-oxidant effect and the ROS scavenging effect. Also RP preserved the systemic anti-oxidant enzyme system (superoxide dismutase and catalase) from UVB irradiation. RP protected the cell membrane from the damages induced by UVB irradiation. RP induced HSP70, a mediator of resistance to UVB irradiation. RP reduced the synthesis of MMP-2 induced by UVB irradiation. And RP inhibited the amount of senescent-associated (SA) ${\beta}-galactosidase$ staining, as a marker of replicative senescence. Conclusions : The results of our study indicate that the extract of the needles of red pine, Pinus densiflora, has anti-aging effects on skin.

• 생명과학회지
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• 제18권6호
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• pp.796-803
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• 2008

알츠하이머 질환은 신경퇴행성질환으로 노령인구에서 뿐만 아니라 $30{\sim}60$세 사이에서도 상염색체성우성형으로 발생하여 사회문제로 대두되고 있으며 발병기전도 명확하게 규명되지 않은 상태이다. 따라서 이 연구에서는 hAPP695sw 돌연변이를 neuron-specific enolase (NSE) 유전자의 프로모터 조절 하에 연결시킨 융합 유전자(pNSE/APP695sw fusion gene)를 과 발현시킨 알츠하이머 질환 모델생쥐를 대상으로 16주간 지구성 운동에 따른 알츠하이머 질환 모델생쥐의 인지능력의 변화와 주병변인 $A{\beta}-42$ 단백질과 함께 GLUT-1, BDNF, HSP-70 단백질의 발현량을 분석하였다. 그 결과 지구성 운동은 APPsw 알츠하이머 질환 모델생쥐의 인지능력을 개선시키는데 긍정적인 영향을 미친 것으로 나타났으며 이러한 인지능력의 개선은 알츠하이머 질환의 주 병변인 뇌의 $A{\beta}-42$ 감소뿐만 아니라 BDNF, GLUT-1과 HSP-70 단백질의 발현 증가와 관련이 있음을 확인하였다. 따라서 지구성 운동은 약물 처치 이외에 알츠하이머 질환을 예방하거나 지연시킬 수 있는 전략적인 방법으로 활용할 수 있음을 알 수 있다.

• Journal of Animal Science and Technology
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• 제52권4호
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• pp.265-270
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• 2010

본 실험은 지방대사에 영향을 주는 후보 유전자의 발현양상을 조사하기 위하여 bovine mesenchymal stem cell을 이용하여 체외에서 지방분화를 유도하였고, 지방분화 경과에 따른 지방관련 후보유전자의 발현양상을 분석 하였다. Oil red-O 염색법을 이용하여 지방 분화양상을 조사한 결과, 지방 분화 단계별(0, 2, 4 및 6일)로 붉게 염색된 지방적이 증가되었음을 확인하였고, 지방대사의 indicator인 PPAR$\gamma$와 FABP4 유전자는 지방분화 0일과 비교하여 6일에는 각각 3.11배와 3.11배가 증가하였다. 또한 지방대사와 관련이 있다고 추측되는 ICER, NOV, HSPB1 및 SDH 유전자의 발현양상을 조사한 결과, 지방분화 동안 ICER 유전자는 계속적으로 증가하여 6일은 0일에 비해 4.12배의 유의적인 증가를 보인 반면, NOV, HSPB1 및 SDH 유전자의 발현양은 0일 이후 계속적으로 감소하여, 6일에서 각각 2.89배, 3.18배 및 2.36배의 감소를 나타냈다. 본 실험에서, 등심조직에서 근내지방도에서 차이를 나타냈던 ICER, NOV, HSPB1 및 SDH 유전자를 보다 구체적이고 직접적으로 세포에서 지방분화를 통해 연관성을 조사했을 때, 지방분화와 직접적으로 관련되어 있는 것으로 나타났으며, 지방 대사를 조절하는 유전자로 추정된다.

• 한국어병학회지
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• 제22권3호
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• pp.293-303
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• 2009

본 연구는 저염분에 의한 둥근전복, Haliotis discus discus의 생리학적 변화를 조사하기 위하여 실시하였다. 생리학적 변화를 조사하기 위하여 생존율, 혈림프수, 항산화효소, Respiratory burst, Phenoloxidase, Lysozyme의 활성 및 HSP 70 mRNA 발현을 측정하였다. 둥근전복은 우선, 급격한 염분 스트레스에 따른 둥근전복의 생존율에서는 25, 30 psu 실험구에서만 48 h 째에 폐사된 개체가 발견되었다. 혈림프 수의 변화는 실험구 대부분이 시간의 경과에 따라 점차 감소하는 경향을 나타내었으나, 3 h째 30 psu에서 급격한 증가가 나타난 후 다른 실험구간의 수치와 유사한 수치로 회복되는 경향을 나타내었다. 항산화 효소인 Superoxide Dismutase (SOD)와 Catalase (CAT) 효소는 25 및 30 psu에서 높은 수치를 나타내어 저염분으로의 변화가 둥근전복의 SOD 및 CAT의 증가를 유발시키는 것으로 나타났다. 한편, Respiratory burst 활성은 실험 기간 동안 큰 변화를 나타내지 않았다. 그러나 면역활성을 나타내는 Phenoloxidase은 염분 감소 의존적으로 감소하는 경향을 나타내어, 면역력의 저하를 유발하는 것으로 나타났다. Lysozyme의 활성에 있어서도 Phenoloxidase 활성의 변화와 같은 저염분 정도에 의존하는 경향은 나타내지 않았으나, 25 psu 48 h째에 급격한 저하를 나타내었다. 저염분의 스트레스를 가하여 나타난 HSP 70 mRNA의 염색성은 가장 낮은 염분농도인 25 psu에서는 지속적으로 관찰되었으나, 33 psu에서는 염색성이 낮게 나타났다. HSP 70 mRNA에 대한 $\beta$-actin mRNA의 발현량은 33 psu에서는 실험기간 중 거의 일정하여 스트레스를 받지 않은 것으로 나타났으며, 25 psu에서는 시간의 경과에 따라 증가하다가 다시 감소하여 회복되는 결과를 나타내었다 이상의 결과로, 저염분 현상은 둥근전복에 있어서 주로 Phenoloxidase 및 Lysozyme의 면역계 활성을 낮추는 작용을 통하여 생존에 악영향을 미칠 가능성이 있는 것으로 추정된다. 또한 저염분에 의해 발생되는 항산화효소는 스트레스의 지표로 써 활용할 수 있을 것으로 여겨진다.