• Title/Summary/Keyword: Binding Protein

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Renal Sympathetic Denervation Induces Acute Myocardial Inflammation through Activation of Caspase-1 and Interleukin-1β (교감신경절제술(RDN) 후 caspase-1과 interleukin-1β 활성화로 인해 유발된 염증성 급성심근손상)

  • Lee, Dong Won;Kim, Il Young;Kwak, Ihm Soo
    • Journal of Life Science
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    • v.28 no.2
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    • pp.240-246
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    • 2018
  • Efferent and afferent sympathetic nerves are closely related to the development of hypertension and heart failure. Catheter-based renal sympathetic denervation (RDN) is implemented as a strategy to treat resistant hypertension. We investigated whether RDN procedure causes inflammatory damage on myocardium in the early phase of sympathetic denervation. Twenty-five female swine were divided into 3 groups: normal control (Normal, n=5), sham-operated control (Sham, n=5), and RDN groups (RDN, n=15). The RDN group was further subdivided into 3 subgroups according to the time of sacrifice: immediately (RDN-0, n=5), 1 week (RDN-1, n=5), and 2 weeks (RDN-2, n=5) after RDN. There were no significant changes in the clinical parameters between the normal control and sham-operated group using contrast-media. In the myocardium, inflammatory cytokines, $IL-1{\beta}$ and $TNF-{\alpha}$ increased at the first week, and then decreased at the second week after RDN. Anti-inflammatory cytokine, IL-10 increased immediately, and then decreased at the second week after RDN. Caspase-1 activity and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) expression increased immediately after RDN until the second week. However, nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing protein 3 (NLRP3) expression did not show any significant differences among the groups. The RDN can cause acute myocardial inflammation through activation of caspase-1 and $IL-1{\beta}$. We should pay attention to protecting against early inflammatory myocardial damage after RDN.

Effects of Sinetrol-XPur on Leptin-Deficient Obese Mice and Activation of cAMP-Dependent UCP-2 (Leptin 유전자 결핍 동물모델을 이용한 시네트롤(Sinetrol-XPur)의 항비만 효과와 cAMP를 통한 UCP-2 활성화 기전 연구)

  • Yoo, Jae Myeong;Lee, Minhee;Kwon, Han Ol;Choi, Sei Gyu;Bae, Mun Hyoung;Kim, Ok-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.484-491
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    • 2016
  • The present study investigated the effect of Sinetrol-XPur (polyphenolic Citrus spp. and Paullinia cupana Kunth dry extract) and defined the action mode for cyclic adenosine monophosphate (cAMP)-dependent uncoupling protein (UCP)-2 activation. Leptin-deficient obese mice were treated with two different doses, 100 mg/kg body weight (BW) and 300 mg/kg BW of each AIN93G supplement, for 7 weeks. Treatment of obese mice with both low and high doses of Sinetrol-XPur significantly reduced body weight gain compared to control obese mice. White adipose tissue weight of mice was reduced by 30.96% in high dose-supplemented groups. Serum total cholesterol and triglyceride were reduced by a high dose of Sinetrol-XPur by 20.02% and 30.96%, respectively. Serum level of high density lipoprotein (HDL) was significantly increased by treatment with both doses, as the ratio of HDL to low density lipoprotein increased by 138.78% and 171.49%, respectively. Regarding expression of biochemical factors related to lipid metabolism, fatty acid synthase significantly decreased and UCP-2 increased upon treatment with a high dose of Sinetrol-XPur, but there was no significant difference in lipoprotein lipase and hormone-sensitive lipase. To define cellular mechanism, intracellular cAMP levels in 3T3-L1 adipocytes significantly increased in a dose-dependent manner over the range of $50{\sim}250{\mu}m/mL$. The phosphodiesterase (PDE) inhibitor 3-isobutyl-1-methylxanthine clearly blocked cAMP, suggesting that Sinetrol-XPur promotes lipolysis of adipocytes through inhibition of cAMP-dependent PDE, resulting in induction of cAMP response element binding protein and UCP-2. These results suggest that Sinetrol-XPur supplementation is a viable option for reducing body weight and fat by improving serum lipid profiles and genetic expression of lipid metabolic factors, especially activation of cAMP-dependent UCP-2.

The Infectivity of Recombinant Porcine Endogenous Retrovirus (PERV-A/C) Is Modulated by Membrane-Proximal Cytoplasmic Domain of PERV-C Envelope Tail (C형 돼지 내인성 레트로바이러스(PERV)의 C-말단 외막당단백질에 의한 재조합 PERV-A/C의 감염력 조절)

  • Kim, Sae-Ro-Mi;Park, Sang-Min;Lee, Kyu-Jun;Lee, Yong-Jin;Bae, Eun-Hye;Park, Sung-Han;Lim, Ji-Hyun;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.15-20
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    • 2010
  • Xenotransplantation of pig organs is complicated by the existence of polytropic replication-competent porcine endogenous retroviruses (PERV) capable of infecting human cells. Two classes of infectious human-tropic replication-competent PERVs (PERV-A and PERV-B) and one class of ecotropic PERV-C are known. The potential for recombination between ecotropic PERV-C and human-tropic PERVs adds another level of infectious risk. A recombinant PERV-A/C (PERV-A14/220) virus is 500-fold more infectious than PERV-A. Two determinants of this high infectivity was identified; one was isoleucine-to-valine substitution at position 140 in RBD (receptor binding domain), and the other lies within the PRR (proline rich region) of the envelope protein. To examine whether the effects of the cytoplasmic tail of the PERV-C Env on fusogenesity also influences infectivity, we constructed a pseudotype retroviral vectors containing MoMLV core protein and PERV envelopes. Pseudotyping experiments with the PERV envelope glycoproteins indicated that recombinant PERV-A/C virus is 10-fold more infectious than PERV-A by lacZ staining. This result supports the suggestion that viral transduction of PERV-A/C is enhanced by a membrane-proximal cytoplasmic amphiphilic ${\alpha}$-helix in PERV-C Env tail.

Activation of the NF-$\kappa$B p50/p65 Complex in Human Lung Cancer Cell Lines (인체 폐암세포주에서 NF-$\kappa$B p50/p65 Complex의 활성화)

  • Choi, Hyung-Seok;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.2
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    • pp.185-194
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    • 1999
  • Background: NF-$\kappa$B is a characteristic transcriptional factor whose functional activity is determined by post-translational modification of protein and subsequent change of subcellular localization. The involvement of the NF-$\kappa$B family of the transcription factors in the control of such vital cellular functions as immune response, acute phase reaction, replication of certain viruses and development and differentiation of cells has been clearly documented in many previous studies. Several recent observations have suggested that the NF-$\kappa$B might also be involved in the carcinogenesis of some hematological and solid tumors. Investigating the possibility that members of the NF-$\kappa$B family participate in the molecular control of malignant cell transformation could provide invaluable information on both molecular pathogenesis and cancer-related gene therapy. Method: To determine the expression patterns and functional roles of NF-$\kappa$B family transcription factors in human lung cancer cell lines NCI-H792, NCI-H709, NCI-H226 and NCI-H157 were analysed by western blot, using their respective antibodies. The nuclear and the cytoplasmic fraction of protein extract of these cell lines were subsequently obtained and NF-$\kappa$B expression in each fraction was again determined by western blot analysis. The type of NF-$\kappa$B complex present in the cells was determined by immunoprecipitation. To detect the binding ability of cell-line nuclear extracts to the KB consensus oligonucleotide, electrophoretic mobility shift assay(EMSA) was performed. Results: In the cultured human lung cancer cell lines tested, transcription factors of the NF-$\kappa$B family, namely the p50 and p65 subunit were expressed and localized in the nuclear fraction of the cellular extract by western blot analysis and immunocytochemistry. Immunoprecipitation assay showed that in the cell, the p50 and p65 subunits made NF-$\kappa$B complex. Finally it was shown by Electrophoretic Mobility Shift Assay(EMSA) that nuclear extracts of lung cancer cell lines are able to bind to NF-$\kappa$B consensus DNA sequences. Conclusion: These data suggest that in human lung cancer cell lines the NF-$\kappa$B p50/p65 complex might be activated. and strengthen the hypothesis that NF-$\kappa$B family transcription factors might be involved in the carcinogenesis of human lung cancer.

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Development of Lotus Root Bugak with Plasma Lipid Reduction Capacity by Addition of Opuntia ficus-indica var. saboten or Green Tea as a Coloring Agent (백년초 및 녹차 가루 첨가 연근 부각의 지질저하 기능성)

  • Kim, Mijeong;Hong, Sun Hee;Chung, Lana;Choe, Eunok;Song, Yeong-Ok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.3
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    • pp.333-340
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    • 2014
  • The purpose of this study was to develop functional lotus root bugak with plasma lipid reduction capacity by controlling the color of batter used for bugak preparation. Lotus root, nearly colorless, was selected to observe color effects. Gardeniae fructus (GF), Opuntia ficus-indica var. saboten (OF), and green tea (GT), which are colored yellow, red, and green, respectively, were used as coloring agents. Fermented glutinous rice was prepared naturally during winter season by placing glutinous rice and water (1:2, w/w) together in a crock pot for 7 days. Coloring materials (10%, w/w) were blended with glue made from fermented glutinous rice flour to prepare the batter. Cooked lotus root was then mixed with a 1.1-fold amount of batter (w/w) and dried at room temperature. Lotus root bugak (LRB) is pan-fried with un-roasted sesame oil, which is traditionally used as frying oil in Korea. Low-density lipoprotein receptor knockout ($LDLr^{-/-}$) mice (n=36) were fed an atherogenic diet (AD) containing various types of LRB (10 g%) for 10 weeks. Plasma triglyceride, total cholesterol, and LDL-C concentrations decreased significantly in mice fed LRB prepared with OF batter (OFB) and GT batter (GTB) (P<0.05). Protein expression levels of fatty acid synthase (FAS) and 3-hydroxyl-3-methylglutaryl coenzyme A reductase (HMGCR) in the OFB and GTB groups were suppressed compared with the LRB group (P<0.05). In accordance with the results on FAS and HMGCR expression, sterol regulatory element binding protein-I and II (SREBP-I and II), which are responsible for the regulation of FAS and HMGCR gene expression, respectively, were down-regulated compared to the LRB group (P<0.05). In conclusion, the plasma lipid reduction activities of OFB and GTB could be mediated through down-regulation of FAS and HMGCR mRNA expression via suppression of regulatory molecules, SREBP-I and II, in $LDLr^{-/-}$ mice.

Basic Studies for the Breeding of High Protein Rice. I. Comparison of the analytical methods for the measurement of the protein content in the brown rice (수두 고단백 계통육성을 위한 기초적 연구 I. 계통육성을 위한 조단백질 분석법의 비교)

  • Mun-Hue Heu;Hak-Soo SUH
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.12
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    • pp.1-5
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    • 1972
  • In order to compare the analytical efficiency of the Kjeldahal, Dye binding and Biurett method for the determination of nitrogen content in the brown rice, correlation coefficients were calculate with the analytical data obtained by the above mentioned 3 different methods for the brown rice of 36 varieties or lines grown at 5 different nitrogen levels (0, 7.5, 15.0, 22.5 and 30.0kg/10a). Analysis of variance were made for the data of 6 varieties among those 36, and compared the precision of the data obtained by the 3 analytical methods. The expenditure (in terms of chemicals and labour) required for the 3 methods are also compared. The results are summarized as follows; 1. The correlation between D. B. C. and Kjeldahl value were generally more significant than the correlation between the value of Biurett and the value of Kjeldahl. But, the D. B. C. method generally overestimates than the Kjeldahl method at both extreme low and extreme high nitrogen contents, and the Biurett method includes more dispersed error than other two methods, though the optical values are parallel to the Kjeldahl nitrogen values at any levels of nitrogen applied. 2. The varietal difference in nitrogen value obtained by the 3 methods were different at the different nitrogen level applied. That is the interaction between variety and analytical method, and between the nitrogen level and analytical method were significant statistically. 3. The coefficient of variance (C, V.) was largest in the data analyzed by Biurett method and next in the data analayred by D. B. C. method. In the data analyzed by Biurett, the C. V. increased along onglong increase of nitrogen applied. But, in the data obtained by D. B. C. or Biurett the C. V. increased along the decrease of nitrogen applied. 4. From the comparison of the expenditure (in terms of chemicals and labour) required for the analysis of 100 samples by 3 methods, it was noticed that, the Biurett or D. B. C. method largely curtail the chemical expenditure and labour costs. Especially the Biurett method could curtail more labour costs.

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Uptake of Butachlor by Rice Seedlings and Its Phytotoxic Action to the Physiological Activities (수도묘(水稻苗)의 Butachlor 흡수(吸收) 및 약해발생(藥害發生) 특성(特性)에 관한 생리적(生理的) 연구(硏究))

  • Chung, Bong-Jin;Kwon, Yong-Woong
    • Korean Journal of Weed Science
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    • v.1 no.1
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    • pp.57-68
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    • 1981
  • To clarify the mode of uptake of butachlor (2-chloro-2', 6'-diethyl-N-(butoxymethyl) acetanilide) by rice seedlings, its phytotoxic action to growth and physiological activities, studies were conducted with rice seedlings, at the 6th or 7th leaf-stage, which were treated with nutrient solution containing butachlor 0, 1.8, 3.6, 7.2, 10.8 or 14.4 ppm for 1, 2 or 4 days, in other case, the solutions were thereafter renewed with the untreated nutrient solution for further growth. Uptake of butachlor by rice seedlings increased linearly with increase of its concentration and duration of uptake. Butachlor inhibited root growth more than shoot growth, furthermore, the inhibitory effect on the shoot growth was greater in height than in weight or leafing rate. After 4 day-treatment, the rates of shoot growth in weight were delayed for 4 days. Butachlor inhibited water uptake rapidly and linearly with increase of its external concentration. The reduced uptake of water was followed by slow increase in the stomatal resistance of leaves. Upon completion of butachlor treatment, rate of water uptake was recovered rapidly, but the stomatal resistance with lag in time. Butachlor did not affect the uptake of cation such as ammonium, potassium and calcium, but inhibited substantially uptake of nitrate in proportion to its concentration. Especially, butachlor did not affect synthesis and degradation of nitrate reductase. In addition, butachlor has shown much greater binding to the lipidic substances from rice roots than the proteinous material. The primary mechanism of phytotoxic action of butachlor does not seem to be its effect on the protein synthesis, but great affinity to membranes. The inhibition of water uptake, and its subsequent closure of stomates is thought very important for reduced growth under mild phytotoxicity.

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Effects of Platycodin D on Gene Expressions of Pro-adipogenic and Anti-adipogenic Regulators in 3T3-L1 Cells (3T3-L1 세포에서 지방세포형성 유도조절자 및 억제조절자의 발현에 대한 platycodin D의 효과)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Cho, Soo-Hyun;Kim, Sung-Su;Kim, Yeong-Shik;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1802-1807
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    • 2009
  • Platycodin D, a major component of Platycodi radix, is known to have various activities including anti-inflammatory, anti-hyperlipidemic, anti-tumor activities and others. Recently, it was reported that platycodin D inhibits fat accumulation and adipogenesis. The aim of this study was to investigate whether various adipogenic regulators are modulated by platycodin D treatment during the adipogenesis of 3T3-L1 cells. mRNA levels of terminal markers of adipogenesis such as ADIPOQ (adiponectin) and GLUT (glucose transporter) 4, which were quantified by real time PCR, were decreased by platycodin D treatment. mRNA expression of PPAR (peroxisome proliferator-activated receptor) $\gamma$ and C/EBP (CCAAT/enhaner binding protein) $\alpha$, which are central transcription factors of adipogenesis, were also decreased by platycodin D treatment. To elucidate the detailed molecular mechanism of platycodin D-induced inhibition of adipogenesis, we analyzed mRNA expression of upstream regulators of PPAR$\gamma$ and C/EPB$\alpha$. mRNA levels of the pro-adipogenic regulators, KROX20 and KLF (Kruppel-like factor) 15 were markedly down-regulated by platycodin D treatment. On the other hand, mRNA expression of CHOP (C/EBP homologous protein), an anti-adipogenic regulator, was significantly up-regulated by platycodin D treatment. mRNA levels of other pro-adipogenic regulators, C/EBP$\beta$ and C/EPB$\delta$, were not affected by platycodin D treatment, and another anti-adipogenic regulator, C/EBP$\gamma$ was also not affected by platycodin D treatment. Taken together, these results suggest that platycodin D-induced inhibition of adipogenesis is mediated by gene interactions including the down-regulation of pro-adipogenic regulators, KROX20 and KLF15, and the up-regulation of an anti-adipogenic regulator, CHOP.

Effects of Garcinia cambogia Extract on the Adipogenic Differentiation and Lipotoxicity (가르시니아 캄보지아 추출물의 지방세포 분화 및 지방 독성에 미치는 영향)

  • Kang, Eun Sil;Ham, Sun Ah;Hwang, Jung Seok;Lee, Chang-Kwon;Seo, Han Geuk
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.411-416
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    • 2013
  • This study aimed to examine the mechanisms underlying the effects of Garcinia cambogia extract on the adipogenic differentiation of 3T3-L1 cells and long-chain saturated fatty acid-induced lipotoxicity of HepG2 cells. 3T3-L1 preadipocytes, mouse embryonic fibroblast-adipose like cell line, were treated with MDI solution (0.5 mM IBMX, 1 ${\mu}M$ dexamethasone, 10 ${\mu}g/mL$ insulin) to generate a cellular model of adipocyte differentiation. Using this cellular model, the anti-obesity effect of Garcinia cambogia extract was evaluated. MDI-induced lipid accumulation and expression of adipogenesis-related genes were detected by Oil red O staining, Nile Red staining, and Western blot analysis. Effects Garcinia cambogia extract on palmitate-induced lipotoxicity was also analyzed by MTT assay, LDH release, and DAPI staining in HepG2 cells. Garcinia cambogia extract significantly suppressed the adipogenic differentiation of preadipocytes and intracellular lipid accumulation in the differentiating adipocytes. Garcinia cambogia extract also markedly inhibited the expression of peroxisome proliferator- activated receptor ${\gamma}2$ ($PPAR{\gamma}2$), CCAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), and adipocyte protein aP2 (aP2). In addition, Garcinia cambogia extract significantly attenuated palmitate-induced lipotoxicity in HepG2 cells. Palmitateinduced cellular damage and reactive aldehydes were also significantly reduced in the presence of Garcinia cambogia extract. These findings suggest that the Garcinia cambogia extract inhibits the adipogenic differentiation of 3T3-L1 preadipocytes, probably by regulating the expression of multiple genes associated with adipogenesis such as $PPAR{\gamma}2$, $C/EBP{\alpha}$, aP2, and thereby modulating fatty acid-induced lipotoxicity to reduce cellular injury in hepatocytes.

The Mechanism of Iron Transport after Intratracheal Instillation of Iron in Rats (랏트의 기관내 Fe 노출후 Fe 이동에 대한 연구)

  • Kwon, Min;Choi, Byung-Sun;Park, Eon-Sub;Chung, Nam-Hyun;Park, Sung-Jo;Lim, Young;Park, Jung-Duck
    • Journal of Preventive Medicine and Public Health
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    • v.37 no.4
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    • pp.329-336
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    • 2004
  • Objectives : Iron (Fe) is an essential element in biological processes; however excessive Fe is harmful to human health. Some air pollutants contain a high level of Fe, and the human lung could therefore be over-exposed to Fe through inhaled air pollutants. This study was performed to investigate the role of metal transporters (divalent metal transporter 1, DMT1, and metal transporter protein 1, MTP1) in the lung under the environments of Fe deficiency in the body and Fe over-exposure in the lung. Methods : Rats were fed Fe deficient (FeD, 2-6 mg Fe/kg) or Fe supplemented (FeS, 120 mg Fe/kg) diet for 4 weeks, followed by a single intratracheal instillation of ferrous sulfate at low (10 mg/kg) or high (20 mg/kg) dose. Fe concentration was analyzed in the serum, lung and liver, and histopathological findings were observed in the lung at 24 hours after Fe administration. The level of DMT1 and MTP1 expression in the lung was analyzed by RT-PCR. Also, the effect of Fe deficiency in the body was evaluated on the level of Fe concentration and metal transporters compared to FeS-diet fed rats at the end of 4-week FeD or FeS diet. Results : The 4-week FeD diet in rats induced an Fe deficiency anemia with decreased serum total Fe, increased unsaturated Fe binding capacity and hypochromic microcytic red blood cells. The concentration of Fe in the lung and liver was lower in the FeD-diet fed rats than in the FeS-diet fed rats. The level of metal transporters mRNA expression was higher in the FeD-diet fed rats than in the FeS-diet. The concentration of Fe in the lung was increased in a dose-dependent pattern after intratracheal instillation of Fe into the rats, while the level of Fe in the serum and liver was not increased in the low-dose Fe administered rats. Therefore, DMT1 and MTP1 mRNA was highly expressed in both FeD-diet and FeS-diet fed rats, after intratracheal instillation of Fe. Conclusions : DMT1 and MTP1 mRNA were more highly expressed in FeD-diet fed rats than in FeS-diet fed rats. The over-exposure of Fe intratracheally induced high expression of metal transporters and increased Fe deposition in the lung in both FeD-diet and FeS-diet fed rats, but did not increase the Fe level of the serum and liver in low-dose Fe administered rats. These results suggest that the role of metal transporters in the lung might be different in a part from the duodenum under the environment of over-exposure to Fe.