아연결핍된 단핵구 U937 Cell Line에 있어서의 유전자 발현 탐색 : cDNA Microarray 기법 이용

Beattie, John H.,Trayhurn, Paul 한국영양학회 2002 Journal of Nutrition and Health Vol.35 No.10

In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

Metallothioneins and Oxidative Stress

John H. Beattie,Paul Trayhurn 한국영양학회 2002 Nutritional Sciences Vol.5 No.4

Metallothioneins and Oxidative Stress

Beattie, John H.,Trayhurn, Paul The Korean Nutrition Society 2002 Nutritional Sciences Vol.5 No.4

The low molecular weight zinc-binding protein metallothionein(MT) contains 32% cysteine and has been shown to efficiently scavenge hydroxyl radicals in vitro. MT expression is induced by oxidative stress and an antioxidant role for this protein has therefore been proposed. This review mainly focuses on the evidence for this role arising from studies using genetically modified animals and cells which either over- or under-express MT. Despite some considerable disparity of results in the literature, reported studies do generally support an antioxidant role. Nevertheless, oxidant stress at non-physiological treatment levels has been the preferred experimental model and there is little information about the role of MT in physiological oxidative stress. Although it is presumed that the mechanism by which MT has an antioxidant effect involves oxidation of cysteinal thiols, it is possible that zinc release from MT is in itself an important signalling factor.

Correlation between Serum Leptin Levels and BMI in Adults Residing in Pohang, Korea

Shin, Hyeon-Soo,Crabtree, Jennifer,Rayner, Vernon,Trayhurn, Paul,Do, Myoung-Sool The Korean Society of Food Science and Nutrition 2005 Preventive Nutrition and Food Science Vol.10 No.1

Leptin is a small polypeptide hormone secreted primarily by adipocytes. Leptin regulates energy balance by decreasing food intake and increasing energy expenditure. This study investigated the relationships between serum leptin levels and BMI (body mass index) in 49 adults in Pohang, Korea. The subjects were 25 males and 24 females, aged 21 to 64 years attending an outpatient clinic at Handong University Sunlin Presbyterian Hospital. Values are given +/- the standard error of the mean. Our study shows that the serum leptin levels in these subjects were positively correlated with BMI. The leptin levels were higher in females (2.39+/-1.82 ng/mL) than in males (0.43+/-0.455ng/mL), although lower than previously reported. We therefore compared the serum leptin levels from the male Korean subjects (BMI 24.3+/-0.74㎏/㎡) with serum from six British males with a similar BMI (23.4+/-1.48㎏/㎡). The serum leptin concentrations (1.76+/-0.76 ng/mL) were lower than that of plasma (4.28+/-1.66 ng/mL) in the British subjects. The serum leptin in the British subjects (1.76+/-0.76ng/mL) was higher than that in the Koreans. There was no correlation between leptin levels and BMI in either male (slope 0.018 ± 0.036, p=0.624) or female (slope 0.382±0.433, p=0.417) type 2 diabetic patients in Pohang, Korea. Taken together, our study shows that the serum leptin level in Koreans varies with the BMI, but is lower than that of BMI-matched British subjects.

Correlation between Serum Leptin Levels and BMI in Adults Residing in Pohang, Korea

Hyeon-Soo Shin,Jennifer Crabtree,Vernon Rayner,Paul Trayhurn,Myoung-Sool Do 한국식품영양과학회 2005 Preventive Nutrition and Food Science Vol.10 No.1

Leptin is a small polypeptide hormone secreted primarily by adipocytes. Leptin regulates energy balance by decreasing food intake and increasing energy expenditure. This study investigated the relationships between serum leptin levels and BMI (body mass index) in 49 adults in Pohang, Korea. The subjects were 25 males and 24 females, aged 21 to 64 years attending an outpatient clinic at Handong University Sunlin Presbyterian Hospital. Values are given +/- the standard error of the mean. Our study shows that the serum leptin levels in these subjects were positively correlated with BMI. The leptin levels were higher in females (2.39+/-1.82 ng/mL) than in males (0.43+/-0.455 ng/mL), although lower than previously reported. We therefore compared the serum leptin levels from the male Korean subjects (BMI 24.3+/-0.74 kg/㎡) with serum from six British males with a similar BMI (23.4+/-1.48 kg/㎡). The serum leptin concentrations (1.76+/ -0.76 ng/mL) were lower than that of plasma (4.28+/-1.66 ng/mL) in the British subjects. The serum leptin in the British subjects (1.76+/-0.76 ng/mL) was higher than that in the Koreans. There was no correlation between leptin levels and BMI in either male (slope 0.018±0.036, p=0.624) or female (slope 0.382±0.433, p=0.417) type 2 diabetic patients in Pohang, Korea. Taken together, our study shows that the serum leptin level in Koreans varies with the BMI, but is lower than that of BMI-matched British subjects.

Inflammatory Gene Expression Patterns Revealed by DNA Microarray Analysis in TNF-α-treated SGBS Human Adipocytes

도명술,정헌순,최봉혁,Leif Hunter,Stuart Langley,Laszlo Pazmany,Paul Trayhurn 연세대학교의과대학 2006 Yonsei medical journal Vol.47 No.5

We report here the use of human inflammation arrays to study the inflammatory gene expression profile of TNF-α- treated human SGBS adipocytes. Human preadipocytes (SGBS) were induced to differentiate in primary culture, and adipocyte differentiation was confirmed, using Oil Red O staining. We treated the differentiated adipocytes with TNF-α, and RNA from differentiated adipocytes with or without TNF-α treatment was hybridized to MWG human inflammation arrays to compare expression profiles. Eleven genes were up- or down-regulated in TNF-α-treated adipocytes. As revealed by array analysis, among 6 up-regulated genes, only eotaxin-1, monocyte chemoattractant protein-1 (MCP-1), and vascular cell adhesion molecule 1 isoform a precursor (VCAM1) were confirmed by real-time polymerase chain reaction (PCR). Similarly, among 5 down-regulated genes, only IL-1 family member 5 (IL1F5), a disintegrin and metalloprotease with thrombospondin motifs-1 preproprotein (ADAMTS1), fibronectin 1 isoform 1 preprotein (FN1), and matrix metalloproteinase 15 preprotein (MMP15) were confirmed by real-time PCR. There was a substantial increase (50-fold) in eotaxin-1 in response to TNF-α. Taken together, we have identified several inflammatory molecules expressed in SGBS adipocytes and discovered molecular factors explaining the relationship between obesity and atherosclerosis, focusing on inflammatory cytokines expressed in the TNF-α-treated SGBS cells. Further investigation into the role of these up- or down-regulated cytokine genes during the pathological processes leading to the development of atherosclerosis is warranted.

Metallothionein Gene Expression in Human Adipocytes of Lean and Obese Subjects

(Myoung Sool Do),(Su Youn Nam),(Sung Eui Hong),(Kyung Wook Kim),(Jacqueline S Duncan),(John H . Beattie),(Paul Trayhurn) 대한비만학회 2002 The Korean journal of obesity Vol.11 No.1