약물유도성 비만의 가능한 분자적 기전

피재은 안양대학교 산업기술연구소 2004 自然科學硏究 Vol.11 No.1

비만을 유발할 수 있는 음식 및 약물의 부작용을 염증반응으로 설명하기 위하여 지방세포(3T3-L1)와 대식세포(RAW 264.7)를 이용하여 그 가능한 분자적 기전을 제시하였다. 본 연구에서는 세포를 재료로 하는 실험으로 음식보다는 표준화된 실험조건을 위하여 비만유발이 대표적 부작용으로 알려진 항정신병약물 olanzapine (OLZ)을 재료로 그 염증반응 결과를 iNOS 및 COX-II의 유전자 발현, 그리고 nitric oxide (NO) 및 prostaglandin (PGE2) 생성정도로 평가하였다. 대식세포와 지방세포에 직접 약물을 처리하였을 때 NO의 대사체인 nitrite농도는 증가하지 않았으나, 약물을 처리한 지방세포의 conditioned media systems을 이용했을 때는 NO 및 PGE2의 생성과 iNOS와 COX-II의 발현 정도가 유의하게 증가하였다. 그러므로 이 결과는 약물부작용으로 유발되는 비만은 염증반응의 하나로 설명할 수 있다. Side effects of some foods and drugs induced the abnormal lipid metabolism have been associated with obesity, which are related with inflammatory reactions. Especially, nitric oxide (NO) and prostaglandin-E2 (PGE2) overproduced in the reactions may be mediated to the obesity. Thus, we hypothesized that drug-induced adverse effects may be related with a low-grade systemic inflammatory reactions. In the present study, we have carried out the experiments that inflammatory mediators such as NO and PGE2can be induced by interactions between adipocyte (3T3-L1) and macrophage (RAW 264.7) after treatment of drug. There was no direct effect of drug on the NO and PGE2 production in both cell types, RAW 264.7 and 3T3-L1. However, the production of NO metabolite and PGE2 increased significantly in RAW 264.7 cells, which were activated with conditioned media from drug-pretreated 3T3-L1 cell. Furthermore, iNOS and COX-II mRNA levels significantly increased in RAW 264.7 cells at early time. Therefore, these results suggest that NO and PGE2 overproduced by drugs can be correlated with the mechanism of obesity.

Correlation between nutrition intake and gene expression profiles in children with asthma

Pie, Jae-Eun,Kim, Yu-Ri,Kim, In-Kyoung,Seo, Sang-Hui,Lee, Seung-Ho,Lee, Hee-Ra,Yoo, Young,Chung, Ji-Tae,Youn, Jong-Pil,Oh, Moon-Ju,Hwang, Seung-Yong,Kim, Meyoung-Kon The Korean Society of Toxicogenomics and Toxicopro 2010 Molecular & cellular toxicology Vol.6 No.3

Asthma is a complex inflammatory disease and its prevalence has increased worldwide, especially in young children. In this study, we used a '24-hour recall method' to identify differences between children with and without asthma in energy intake and energy-adjusted nutrition intake. We also performed reverse transcription-polymerase chain reaction (RT-PCR) with pathway-targeted arrays (RT2 $Profiler^{TM}$ PCR Array) to investigate the expression profiles of chemokines and cytokines in children with asthma. The intake of vitamin C in mild and moderate asthma was significantly higher than that in healthy controls. Vitamin E intake in the mild asthma group was also significantly higher. Twenty-three genes were expressed at higher levels in severe asthma compared with healthy controls. Using the human Th1-Th2-Th3 PCR Array, we found 17 genes were upregulated in severe asthma, including the Th2-related genes CCL7, IL13, and CCL-11 (eotaxin). These PCR Array results revealed that the genes that were most profoundly increased in asthma encoded for key proinflammatory and chemotactic molecules. Our observations lead us to speculate that the interaction between gene expression and dietary intake is important for the development of asthma.

Tissue-Specific Expression of Nitric Oxide Synthase in Nutrient Digestive Organs

Pie, Jae-Eun 안양대학교 산업기술연구소 2001 自然科學硏究 Vol.8 No.-

Using the human samples of trauma patients (IRB approved), differential expression levels of mRNAs and proteins of nitric oxide synthase (NOS) were determined in several digestive organs, including small and large intestines. Two different sizes (^~11 kb in testis, small intestine and colon, and ^~12 kb in other tissues examined) of nNOS mRNA were detected in tissue specific patterns. In the Western-blot analysis, however, at least three different nNOS bands (130, 140, and 160 kDa) were observed, particularly in small intestine and colon (160 and 140 kDa). And two iNOS mRNAs of different sizes (4.5 and 4.2 kb) were also expressed constitutively at different levels in peripheral tissues including small intestine and colon. In some tissues, however, only a single band iNOs protein was observed. As with the iNOS mRNAs, two eNOS mRNAs of different sizes (4.4 and 4.2 kb) were detected in almost all tissues examined. These tissue-specific or differential expression of NOS mRNAs and proteins are produced by alternative splicing and this may be associated with the diverse tissue specific roles of NO observed in vivo, especially in digestive organs.

Positive Correlation between Suppressed Brug Metabolizing Enzymes and Increased iNOS Expression in Mouse Liver after Schistosoma japonicum Infection

Pie, Jae-Eun 안양대학교 산업기술연구소 2000 自然科學硏究 Vol.7 No.-

Activities of two major hepatic drug metabolizing enzymes, cytochrome P450(CYP) and flavin-containing monooxygenase (FMO) were examined in mice after a moderate infections of S. japonicum. Liver microsomal CYP activities determined by testosterone hydroxylation and caffeine oxidation and FMO activities analyzed by ranitidine N-oxidation were significantly decreased by 6 weeks but recovered in 9 weeks with significant difference between male and female. Conversely. the hepatic iNOS mRNA contents increased maximally at 6-7 weeks but decreased toward normal undetectable level by 9 weeks. Immunohistochemical detection of iNOS expression sites showed initial moderate staining at Kupffer cells, intense staining of granuloma cells forming a halo surrounding the deposited eggs (6-7 weeks) and dark focal staining only in the core of dying parasite eggs (9 weeks). Results suggested that the NO produced by iNOS and the super-oxide anion (02-) produced by the granulomatous cells may generate the toxic peroxynitrite (ONOO-). This, in turn, may mediate the suppression of the drug metabolism by inhibiting the CYP and FMO activities in hepatocytes.

Nitric Oxide Dependency in Inflammatory Response-related Gene Transcripts Expressed in Lipopolysaccharide-treated RAW 264.7 Cells

Pie, Jae-Eun,Yi, Hyeon-Gyu The Korean Society of Toxicogenomics and Toxicopro 2009 Molecular & cellular toxicology Vol.5 No.4

Cytotoxic Nitric oxide (NO) overproduced by inducible NO Synthase (iNOS or NOS2), which was induced in inflammatory reactions and immune responses directly or indirectly affects the functions as host defense and can cause normal tissue damage. Microarray analysis was performed to identify gene profiles of both NO-dependent and -independent transcripts in RAW 264.7 macrophages that use selective NOS2 inhibitors aminoguanidine ($100\;{\mu}M$) and L-canavanine (1 mM). A total of 3,297 genes were identified that were up- or down-regulated significantly over 2-fold in lipopolysaccharide (LPS)-treated macrophages. NO-dependency was determined in the expressed total gene profiles and also within inflammatory conditions-related functional categories. Out of all the gene profiles, 1711 genes affected NO-dependently and -independently in 567 genes. In the categories of inflammatory conditions, transcripts of 16 genes (Pomp, C8a, Ifih1, Irak1, Txnrd1, Ptafr, Scube1, Cd8a, Gpx4, Ltb, Fasl, Igk-V21-9, Vac14, Mbl1, C1r and Tlr6) and 29 geneas (IL-1beta, Mpa2l, IFN activated genes and Chemokine ligands) affected NO-dependently and -independently, respectively. This NO dependency can be applied to inflammatory reaction-related functional classifications, such as cell migration, chemotaxis, cytokine, Jak/STAT signaling pathway, and MAPK signaling pathway. Our results suggest that LPS-induced gene transcripts in inflammation or infection can be classified into physiological and toxic effects by their dependency on the NOS2-mediated NO release.

열무 물김치의 담금방법이 발효숙성에 미치는 영향

피재은(Jae-Eun Pie),장명숙(Myung-Sook Jang) 한국식품영양과학회 1995 한국식품영양과학회지 Vol.24 No.6

열무 물김치의 담금방법이 발효숙성에 미치는 영향을 알아보기 위하여 이화학적 특성(pH, 총 산 함량, 환원당, 총 비타민 C, 색도), 미생물학적 특성과 관능적 특성을 조사하였다. 열무 물김치는 4가지 담금방법인 밀가루풀과 고춧가루를 넣지 않는 방법(A), 밀가루풀만을 넣는 방법(B), 고춧가루만을 넣는 방법(C)과 밀가루풀과 고춧가루를 함께 넣는 방법(D)으로 하여 모든 시료는 실온(27±0.5℃)에서 8시간 숙성 후 4℃에 각각 20일 동안 발효숙성시켰다. pH는 발효가 진행됨에 따라 모두 점차적으로 낮아지는데, 발효 10일 이후에는 변화정도가 완만하게 낮아짐을 볼 수 있었다. 발효 10일까지는 대체적으로 시료구 D가 약간 높은 pH를 나타내었으나, 10일 이후에는 오히려 시료구 B가 약간 낮은 pH를 나타내었다. 총 산 함량은 발효의 진행과 함께 증가함을 보였다. 환원당은 초기에 약간 증가하다가 서서히 감소하는 경향을 나타내었다. 특히 시료구 B는 발효의 진행과 함께 증가하였다가 발효 20일에도 거의 초기함량을 유지하는 경향을 나타내었다. 총 비타민 C는 전체적으로 발효 20일 동안 초기 함량 보다 점차로 감소하는 경향을 보였다. 시료구 D는 다른 시료구에 비해서 초기에 총 비타민 C 함량이 증가한 것을 볼 수 있었다. 색도에서 명도와 황색도는 큰 변화를 보이지 않았지만, 적색도는 모두 증가하는 경향을 나타내었다. 총 균수와 젖산균수의 변화는 총 균수가 젖산균수 보다 많은 것을 제외하고는 발효의 진행에 따라 거의 비슷하였는데, 발효 10일에서 13일 사이에 모두 최대값을 보이고 그 이후에 모든 시료에서 서서히 감소하였다. 시료구 D가 발효 전체 기간 동안 가장 많은 총 균수를 나타내었고, 시료구 A가 가장 적은 총 균수를 나타내었다. 관능적평가에서는 모두 유의적인 차이를 나타내었고, 모든 항목에서 발효 10일 전에는 시료구 D가 비교적 높은 점수를 받았다. 반면에 10일 이후에는 시료구 B가 높은 점수를 받았다. Effect of preparation method on Yulmoo kimchi(watery kimchi prepared with Yulmoo, leaf radish) fermentation was investigated by measuring physicochemical, microbiological and sensory properties for 20 days. Yulmoo kimchi was fermented at 4℃ after keeping at room temperature(27±0.5℃) for 8 hours. Four types (A, B, C, D) of Yulmoo kimchi were prepared. Sample A was made without wheat flour paste and red pepper powder and sample B was made with wheat flour paste. Sample C was made with red pepper powder, and sample D was made with wheat flour paste and red pepper powder. pH was slowly lowered and stabled after 10 days in all samples. Total acid content increased in all samples. Reducing sugar content initially increased and decreased thereafter. Especially, reducing sugar content of sample B increased in the early stage of fermentation and maintained initial reducing sugar content after 20 days. Total vitamin C content reduced during fermentation in all samples. Especially initial total vitamin C content of sample D increased more than those of other samples relatively. Lightness and yellowness showed no difference, but on the other hand redness increased gradually in all samples. The number of lactic acid bacteria reached maximum value in 10~13 days with the total cell numbers and gradually decreased thereafter in all samples. The number of lactic acid bacteria and total cell number of sample D was much more than those of any other samples. As a result of the sensory evaluation, Yulmoo kimchi showed siginificant difference in all characteristics. Sample D showed the highest scores in all characteristics before 10 days of fermentation. However, after 10 days sample B showed the highest scores.

단백질의 종류가 체내 칼슘대사에 미치는 영향에 관한 연구

피재은(Pie Jae-Eun),백희영(Paik Hee Young) 韓國營養學會 1986 Journal of Nutrition and Health Vol.19 No.1

Effects of Jack Bean Extract on human articular chondrocytes

피재은 안양대학교 산업기술연구소 2003 自然科學硏究 Vol.10 No.-

Differential Diagnosis of Chemical-induced Hepatobiliary Toxicities Using a New Hepatobiliary Imaging Agent in Mice

Chong-Kun Ryu,Jae-Eun Pie,Jae Gol Choe,Joon Cheon,Jeongwon Sohn,Jurgen Seidel,David S. Paik,Michael V. Green,Chang H. Paik,Meyoung-Kon Kim 한국환경성돌연변이발암원학회 2001 한국환경성돌연변이·발암원학회지 Vol.21 No.1

We have synthesized 99mTc-mercaptoacetyltriglycine (MAG3)-biocytin as a new imaging agent for hepatobiliary scintigraphy. The aim of this study was to evaluate the usefulness of 99mTc-MAG3-biocytin scintigraphy in differentiating carbon tetrachloride (CCl₄)-induced hepatotoxicity from a-naphthylisothiocyanate (ANIT)-induced cholestasis in mice, which reflecting the differential diagnosis of neonatal jaundice caused by neonatal hepatitis from congenital biliary atresia in humans. Methods: Balb/c mice (female, 20 g, n=4-6) were pretreated with CCl4 (0.5 or 1.0 ml/kg) and ANIT (150 or 300 mg/kg) 18 h before scintigraphy. Biochemical and histopathological examinations showed a pattern of typical acute hepatitis (increase of transaminases and hepatocellular necrosis) in CCl4-treated mice and cholestasis (increase of alkaline phosphatase and g-glutamyltransferase, and biliary hyperplasia) in ANIT-treated mice, respectively. Mice were fasted at least 4 hr prior to the intravenous injection of 99mTc-MAG3-biocytin (18.5 MBq/ 20㎍) in 2% human serum albumin in saline. Scintigraphy<br/> was performed with a g-camera equipped with a 1-mm diameter pin-hole collimator for 30 min and images were acquired every 15 s. We compared the values of physical parameters, such as peak liver/heart ratio (rmax) and peak ratio time (tmax) for 99mTc-MAG3-biocytin scintigraphy. Results: Scintigraphic parameters of the<br/> CCl₄-pretreated (0.5 ml/kg) group showed a 81.9% decrease of rmax, and 42.2% decrease of tmax, whereas the ANIT-pretreated (150 mg/kg) group showed a 53% decrease of rmax, and 2.36-fold increase of tmax, (P<0.05). These results demonstrate that the decrease of rmax and the shortening of tmax are characteristic features for<br/> hepatotoxicity, in contrast to the increase of tmax and decrease of rmax for biliary hyperplasia. Conclusion: 99mTc-MAG3-biocytin hepatobiliary scintigraphy can distinguish hepatitis from cholestasis in mice model and may be similarly useful in humans which differentiating the cause of neonatal jaundice in clinical study.

Effects of zinc oxide nanoparticles on gene expression profile in human keratinocytes

Lee, Seung Ho,Pie, Jae-Eun,Kim, Yu-Ri,Lee, Hee Ra,Son, Sang Wook,Kim, Meyoung-Kon 대한독성유전단백체학회 2012 Molecular & cellular toxicology Vol.8 No.2

Zinc oxide (ZnO) nanoparticle is added in various materials and products such as paints, plastics, ceramics, glass, rubber, pigments and sunscreen. Especially, ZnO nanoparticles are widely used in UV protection, because of their property of absorption ultraviolet light. Despite the widespread use of ZnO nanoparticles, there are many unknowns in understanding on their nanotoxicity and mechanisms. In this study, we evaluated the effects of ZnO nanoparticles on gene expression pattern of human keratinocyte cells. Total RNA was prepared from the exposure groups to ZnO nanoparticles with different surface charge, and cDNA microarray was performed using Agilent human whole genome array. Our study indicated that genes related to apoptosis and response to stress including heme oxygenase 1 (HMOX1), superoxide dismutase (SOD), glutathione peroxidase (GPX), BNIP3L (adenovirus E1B 19 kDa interacting protein 3) and heat shock 70 kDa protein (HSP70) were up- and down-regulated in ZnO nanoparticles treated cells. Moreover, our results showed that ZnO nanoparticles induced intracellular reactive oxygen species (ROS) and oxidative stress. Antioxidant enzyme SOD levels were significantly higher and GSH levels were decreased in ZnO nanoparticles-exposed cells, respectively. The present study showed that up-regulation of these genes by ZnO nanoparticles could increase the production of ROS and oxidative stress. Therefore, ZnO nanoparticles could have a potential to product ROS through the perturbation of metabolic pathway, inducing oxidative stress. It also supported that the nanotoxicity mechanism could correlate with the active oxygen production, oxidative stress, apoptosis, and antioxidant defense mechanisms.