The Hepatic Raldh1 Expression Is elevated in Zucker Fatty Rats and Its Over-Expression Introduced the Retinal-Induced Srebp-1c Expression in INS-1 Cells
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{"title"=>"The Hepatic Raldh1 Expression Is elevated in Zucker Fatty Rats and Its Over-Expression Introduced the Retinal-Induced Srebp-1c Expression in INS-1 Cells", "type"=>"journal", "authors"=>[{"first_name"=>"Yang", "last_name"=>"Li", "scopus_author_id"=>"55922462300"}, {"first_name"=>"Yan", "last_name"=>"Zhang", "scopus_author_id"=>"56216293000"}, {"first_name"=>"Rui", "last_name"=>"Li", "scopus_author_id"=>"57189517648"}, {"first_name"=>"Wei", "last_name"=>"Chen", "scopus_author_id"=>"57087492200"}, {"first_name"=>"Meredith", "last_name"=>"Howell", "scopus_author_id"=>"55360751100"}, {"first_name"=>"Rui", "last_name"=>"Zhang", "scopus_author_id"=>"55613242686"}, {"first_name"=>"Guoxun", "last_name"=>"Chen", "scopus_author_id"=>"7406542711"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"365666241", "doi"=>"10.1371/journal.pone.0045210", "sgr"=>"84866419847", "scopus"=>"2-s2.0-84866419847", "pmid"=>"23028851"}, "id"=>"433a051b-27bd-3d71-984a-62eb32e536b1", "abstract"=>"The roles of vitamin A (VA) in the development of metabolic diseases remain unanswered. We have reported that retinoids synergized with insulin to induce the expression of sterol-regulatory element-binding protein 1c gene (Srebp-1c) expression in primary rat hepatocytes. Additionally, the hepatic Srebp-1c expression is elevated in Zucker fatty (ZF) rats, and reduced in those fed a VA deficient diet. VA is metabolized to retinoic acid (RA) for regulating gene expression. We hypothesized that the expression of RA production enzymes contributes to the regulation of the hepatic Srebp-1c expression. Therefore, we analyzed their expression levels in Zucker lean (ZL) and ZF rats. The mRNA levels of retinaldehyde dehydrogenase family 1 gene (Raldh1) were found to be higher in the isolated and cultured primary hepatocytes from ZF rats than that from ZL rats. The RALDH1 protein level was elevated in the liver of ZF rats. Retinol and retinal dose- and time-dependently induced the expression of RA responsive Cyp26a1 gene in hepatocytes and hepatoma cells. INS-1 cells were identified as an ideal tool to study the effects of RA production on the regulation of gene expression because only RA, but not retinal, induced Srebp-1c mRNA expression in them. Recombinant adenovirus containing rat Raldh1 cDNA was made and used to infect INS-1 cells. The over-expression of RALDH1 introduced the retinal-mediated induction of Srebp-1c expression in INS-1 cells. We conclude that the expression levels of the enzymes for RA production may contribute to the regulation of RA responsive genes, and determine the responses of the cells to retinoid treatments. The elevated hepatic expression of Raldh1 in ZF rats may cause the excessive RA production from retinol, and in turn, result in higher Srebp-1c expression. This excessive RA production may be one of the factors contributing to the elevated lipogenesis in the liver of ZF rats", "link"=>"http://www.mendeley.com/research/hepatic-raldh1-expression-elevated-zucker-fatty-rats-overexpression-introduced-retinalinduced-srebp1", "reader_count"=>8, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Other"=>1, "Unspecified"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>1, "Other"=>1, "Unspecified"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>2, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Neuroscience"=>{"Neuroscience"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/577300"], "description"=>"<p><b>A and B.</b> Total RNA was extracted from hepatocytes and subjected to real-time PCR analysis. Results were presented as means ± SD of the −ΔC<sub>T</sub> (the C<sub>T</sub> of 36B4– the C<sub>T</sub> of indicated transcript) from the indicated different numbers (in parenthesis) of hepatocyte isolations (* all <i>p</i><0.05, for comparing the values of ZL and ZF groups of the indicated transcripts using the independent t-test). <b>C.</b> Whole liver tissue lyastes (100 µg/lane) of ZL (lanes 1–4) and ZF (lanes 5–8) rats, and whole cell lysate of INS-1 cells infected by Ad-Raldh1 (50 µg, lane 9) were separated in 10% SDS PAGE gels, and transferred to the PVDF membranes as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045210#s4\" target=\"_blank\">Material and Methods</a>. Primary antibodies to RALDH1 (1∶1000 dilution in TBST containing 5% dry milk), and to β-Actin (1∶1000 dilution in TBST containing 5% bovine serum albumin) were recognized by goat anti-rabbit IgG conjugated to horseradish peroxidase, and visualized by chemiluminescence. The ratios of RALDH1/β-Actin were calculated after the films were scanned and analyzed. Data were presented as mean ± SD of the indicated numbers (in parenthesis) of ZL and ZF rats (** <i>p</i><0.04 for comparing the ratios of ZL rats with that of ZF rats using the independent t-test).</p>", "links"=>[], "tags"=>["mrna", "levels", "freshly", "cultured", "raldh1", "zl", "zf"], "article_id"=>247776, "categories"=>["Molecular Biology", "Biochemistry"], "users"=>["Yang Li", "Yan Zhang", "Rui Li", "Wei Chen", "Meredith Howell", "Rui Zhang", "Guoxun Chen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0045210.g001", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_mRNA_levels_of_Rdh2_and_Raldh1_in_freshly_isolated_A_and_cultured_B_primary_hepatocytes_and_the_RALDH1_protein_levels_C_in_the_liver_of_ad_libitum_ZL_and_ZF_rats_/247776", "title"=>"The mRNA levels of <i>Rdh2,</i> and <i>Raldh1</i> in freshly isolated (A) and cultured (B) primary hepatocytes, and the RALDH1 protein levels (C) in the liver of <i>ad libitum</i> ZL and ZF rats.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:23:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/577387"], "description"=>"<p>Primary hepatocytes were isolated from SD rats, and pre-treated as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045210#s4\" target=\"_blank\">Materials and Methods</a>. Total RNA was isolated, and subjected to real-time PCR analysis. <b>A.</b> A 24-h time course of <i>Cyp26a1</i> mRNA expression induced by RA (5 µM) in primary rat hepatocytes (* for comparing the vehicle control with the RA groups at the indicated time points using the independent t-test; ** and '' for comparing the different time points of vehicle control or RA group using one way ANOVA; all <i>p</i><0.05). <b>B.</b> The expression levels of <i>Cyp26a1</i> mRNA in primary hepatocytes treated with increasing concentrations of ROL or RAL for 6 hours ($ for comparing the fold induction of ROL group with that of RAL group at the indicated concentrations using the independent t-test; c > b > a; h > g > f > e > d for comparing the different dosages of ROL or RAL using one way ANOVA; all <i>p</i><0.05). <b>C.</b> Inhibition of the RAR activation disrupted the induction of <i>Cyp26a1</i> by RAL treatment. Primary hepatocytes were incubated in medium A without or with 2 μM RAL in the absence or presence of increasing concentrations of Ro 41–5253 for 6 hours (i/j > l, i'/j' > k' > l' > m' for comparing the different dosages of Ro 41–5253 in the absence or presence of RAL using one way ANOVA, all <i>p</i><0.05). Results were presented as means ± SD of three independent hepatocyte isolations.</p>", "links"=>[], "tags"=>["levels", "mrna", "ra", "24-h", "dosages", "rol", "ral", "antagonist", "rar", "activation"], "article_id"=>247867, "categories"=>["Molecular Biology", "Biochemistry"], "users"=>["Yang Li", "Yan Zhang", "Rui Li", "Wei Chen", "Meredith Howell", "Rui Zhang", "Guoxun Chen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0045210.g002", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_expression_levels_of_Cyp26a1_mRNA_in_response_to_RA_treatment_over_a_24_h_time_period_A_to_different_dosages_of_ROL_or_RAL_B_and_to_an_antagonist_of_the_RAR_activation_C_/247867", "title"=>"The expression levels of <i>Cyp26a1</i> mRNA in response to RA treatment over a 24-h time period (A), to different dosages of ROL or RAL (B), and to an antagonist of the RAR activation (C).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:24:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/577553"], "description"=>"<p>A. 834/40 INS-1 cells were treated with increasing concentrations of RAL or RA for 6 h. <b>B.</b> 834/40 cells were treated with 2 µM of RAL or RA for indicated time. <b>C.</b> 833/15 cells were treated with 1 µM of specific ligands of RXR (LG268), RAR (TTNPB) or in combination. Total RNA was extracted from INS-1 cells and subjected to real-time PCR analysis. Results were presented as means ± SD of fold inductions (c > b > a for comparing the different dosages of RA-treated groups using one way ANOVA, and e > d > f for comparing control with LG 268 in the absence or presence of TTNPB using one way ANOVA; * for comparing RAL with RA groups at the indicated dosages, and ** for comparing RA with RAL or vehicle control groups at the indicated time points using independent t-test; <i>n</i> = 3, all <i>P</i><0.05).</p>", "links"=>[], "tags"=>["mrna", "levels", "ins-1", "cells", "treated", "retinoids"], "article_id"=>248041, "categories"=>["Molecular Biology", "Biochemistry"], "users"=>["Yang Li", "Yan Zhang", "Rui Li", "Wei Chen", "Meredith Howell", "Rui Zhang", "Guoxun Chen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0045210.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_Srebp_1c_mRNA_levels_in_INS_1_cells_treated_with_retinoids_or_ligands_/248041", "title"=>"The <i>Srebp-1c</i> mRNA levels in INS-1 cells treated with retinoids or ligands.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:25:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/577470"], "description"=>"<p>The dose-dependent induction of <i>Cyp26a1</i> by ROL, RAL, and RA. HL1C cells were treated with ROL, RAL, or RA at the doses of 0, 0.002 µM, 0.02 µM, 0.2 µM, 2 µM, and 20 µM for 6 h (a/b < c, d/e < f, g/h < i, j < k/l, m < n/o, a/d/g/j < m, b/e/h < k < n, c/f < i/l/o for comparing ROL, RAL and RA at the indicated concentrations, and for comparing each ligand at the different dosages using one way ANOVA; all <i>p</i><0.05). <b>B.</b> The expression levels of <i>Cyp26a1</i> mRNA overtime (3, 6, 9, 12, and 24 h) in HL1C cells treated with 5 µM RAL or RA (* or ** for comparing RAL or RA with vehicle control groups at the indicated time points using the independent t-test; all <i>p</i><0.05). <b>C.</b> The expression levels of <i>Cyp26a1</i> mRNA in HL1C cells treated with RAR (TTNPB), RXR (LG268), and LXR (T1317) agonists. HL1C cells were treated with 1 μM TTNPB, LG268, and T1317 for 6 h (q > r>s using one way ANOVA; all <i>p</i><0.05). Total RNA was extracted and subjected to real-time PCR analysis. Data were presented as mean ± SD of three independent treatments. <b>D.</b> Whole cell lysates (50 µg/sample) of HL1C cells treated the vehicle control, 5 µM RAL or RA for 12 (lanes 1–3) and 24 (lanes 4–6) hours were separated in 8% SDS PAGE gels, and transferred to the PVDF membranes. Primary antibodies to CYP26A1 (1∶1000 dilution in TBST containing 5% dry milk), and to β-Actin (1∶1000 dilution in TBST containing 5% bovine serum albumin) were recognized by goat anti-rabbit IgG conjugated to horseradish peroxidase, and visualized by chemiluminescence. The films were scanned and presented as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045210#s4\" target=\"_blank\">Material and Methods</a>.</p>", "links"=>[], "tags"=>["retinoids", "levels", "mrna", "cyp26a1", "hl1c"], "article_id"=>247956, "categories"=>["Molecular Biology", "Biochemistry"], "users"=>["Yang Li", "Yan Zhang", "Rui Li", "Wei Chen", "Meredith Howell", "Rui Zhang", "Guoxun Chen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0045210.g003", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_effects_of_retinoids_on_the_expression_levels_of_Cyp26a1_mRNA_and_CYP26A1_protein_in_HL1C_cells_A_/247956", "title"=>"The effects of retinoids on the expression levels of <i>Cyp26a1</i> mRNA and CYP26A1 protein in HL1C cells. A.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:24:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/577607"], "description"=>"<p>The adenovirus-mediated <i>Raldh1</i> mRNA expression. <b>B.</b> Immuno-blot of the over-expression of RALDH1 protein in INS-1 cells. Whole cell lysates (50 µg/sample) of the control cells (lane 1), cells infected by the indicated <i>pfu</i> of Ad-β-gal (lane 2) or Ad-Raldh1 (lanes 3–5) were separated in 8% SDS protein gels, and transferred to the PVDF membranes. Primary antibodies to RALDH1 (1∶1000 dilution in TBST containing 5% dry milk), and to β-Actin (1∶1000 dilution in TBST containing 5% bovine serum albumin) were recognized by goat anti-rabbit IgG conjugated to horseradish peroxidase, and visualized by chemiluminescence. The films were scanned and presented as described in the <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045210#s4\" target=\"_blank\">Material and Methods</a>. <b>C.</b> RAL only induced <i>Srebp-1c</i> expression in cells over-expressing RALDH1, but not β-gal. <b>D.</b> RA induced <i>Srebp-1c</i> expression in cells over-expressing either β-gal or RALDH1. Results were presented as means ± SD of fold inductions (* for comparing the different dosages of RAL in cells infected by Ad-Raldh1 using one way ANOVA, <i>n</i> = 3, all <i>p</i><0.05).</p>", "links"=>[], "tags"=>["over-expression", "raldh1", "resulted", "ral-mediated", "induction", "ins-1"], "article_id"=>248092, "categories"=>["Molecular Biology", "Biochemistry"], "users"=>["Yang Li", "Yan Zhang", "Rui Li", "Wei Chen", "Meredith Howell", "Rui Zhang", "Guoxun Chen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0045210.g005", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_over_expression_of_RALDH1_resulted_in_the_RAL_mediated_induction_of_Srebp_1c_in_833_15_INS_1_cells_A_/248092", "title"=>"The over-expression of RALDH1 resulted in the RAL-mediated induction of <i>Srebp-1c</i> in 833/15 INS-1 cells. A.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:25:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/577677"], "description"=>"<p><b>A.</b> In ZL rats, the hepatic expression of <i>Srebp-1c</i> is controlled by the RA production and insulin stimulation. The SREBP-1c also regulates RA production via the induction of <i>Raldh1</i> expression until a homeostasis is reached. <b>B.</b> The hyperphagia of ZF rats due to the leptin receptor deficiency causes the over-supply of dietary VA, and hyperinsulinemia. The possible elevation of RA production in combination with insulin stimulation leads to higher expression of hepatic <i>Srebp-1c</i> in ZF rats, which disrupts the homeostasis. The expression of <i>Raldh1</i> mRNA is further induced by SREBP-1c. Therefore, the hepatic expression of <i>Srebp-1c</i> in ZF rats is maintained at a higher level than that in ZL rats. The consequence is the elevated hepatic lipogenesis in ZF rats. The up arrows next to the texts indicate induction. The intensified weight of the lines indicates the induction of that part of the pathway. The question marks indicate the steps remained to be confirmed in this hypothesis.</p>", "links"=>[], "tags"=>["hypothesized", "ra", "feed-forward", "induction", "downstream", "lipogenic", "genes", "zl", "zf"], "article_id"=>248161, "categories"=>["Molecular Biology", "Biochemistry"], "users"=>["Yang Li", "Yan Zhang", "Rui Li", "Wei Chen", "Meredith Howell", "Rui Zhang", "Guoxun Chen"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0045210.g006", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_hypothesized_role_of_the_RA_production_in_the_feed_forward_induction_of_the_expression_of_Srebp_1c_and_its_downstream_lipogenic_genes_in_the_liver_of_ZL_A_and_ZF_B_rats_/248161", "title"=>"The hypothesized role of the RA production in the feed-forward induction of the expression of <i>Srebp-1c</i> and its downstream lipogenic genes in the liver of ZL (A) and ZF (B) rats.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:25:54"}

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