Influence of Low Dose-Rate Radiation on Metabolism

Takashi SUGIHARA, Katsuyoshi FUJIKAWA, Satoshi TANAKA, Ignacia TANAKA, Jun-ichiro KOMURA Department of Radiobiology

Abstract To elucidate the influence of low dose-rate (LDR) radiation on metabolism, we have started quantitative analysis of the amount of low molecular-weight metabolites (metabolome analysis) in the liver of B6C3F1 male and female mice irradiated at a LDR of 20 mGy/day for 300 days (total dose: 6,000 mGy). Here we report preliminary results obtained this year. We observed changes in the amount of the compounds involved in the TCA cycle, gluconeogenesis and glycolysis (for example, fumaric acid, malic acid, glyceraldehyde triphosphate and phosphoenolpyruvate) in both male and female irradiated mice. These results suggest that metabolome analysis can be a useful tool for analyzing the biological effects of LDR radiation.

Non-irradiated Irradiated Non-irradiated Irradiated

Fig.1. Score plot of principal component analysis of the metabolites in the liver of male and female mice irradiated or non-irradiated at 20 mGy/day for 300 days. n = 4. 0.15

Asn Type2

His

Galactosamine Phe Glucosamine Ascorbate 2-glucoside Hexanoic acid Lys S-Methylglutathione Xanthosine Adenine Pro Gln 0.1 Cysteine glutathione disulfide Isobutyryl CoA_divalent

Glycerol 3-phosphate Leu Ala Trimethylamine Tyr XC0126 Type1 Gly Isovaleric acid Guanidinosuccinic acid Type1 Daminozide Ile GDP-glucose Trp Heptanoic acid Octanoic acid Valeric acid Ala-Ala GDP-mannose 1-Methyladenosine Pyridoxamine 5'-phosphate Xanthine GDP-galactose Thr PenciclovirInosine Riboflavin Ser Nicotinamide Octanoylcarnitine Val dTDP-glucoseTaurocholic acid Octanoyl CoA_divalent Picolinic acid Homocitrulline Isovalerylcarnitine GDP N6-AcetyllysineUric acid Putrescine 6-Phosphogluconic acid N-Acetylglutamine UDP-N-acetylgalactosamine-1 Spermidine UDP S-Lactoylglutathione UDP-N-acetylglucosamine-1 Uridine Anthranilic acid IsethionicGlucose 1 acid-phosphate Gulonolactone Taurine Glucose 6-phosphate Lactic acid Thiamine GABA Ribulose 5-phosphate Dyphylline Thiaproline 0.05 XC0071 Glycerol Acetyl CoA_divalent N-Acetylalanine N6-Methyl-2'-deoxyadenosine Carnosine 2-Hydroxyisobutyric acid ThrPyrophosphate-Asp CystathionineOrnithineHypotaurine Quinolinic acid Metronidazole AMPUTP GTP Sarcosine GlutathioneGlycerophosphocholine (GSH) Aminoacetone2Acetoacetamide-Phosphoglyceric acid 3-Ureidopropionic acid β-Ala-Lys Cys 2-1Methylserine-Aminocyclopropanep-Toluic acid6-Hydroxyhexanoic-1-carboxylicUracilIMP acidacid-2 Tyr-Arg_divalent XA0055 Arg Rhein dIMP Homoserinelactone-2 2-Amino-3-phosphonopropionic acid Hexylamine m-Toluic acid Threonic acid N1Butyrylcarnitine-Acetylspermidine Piperidine 3-AminobutyricN1-Methyl acid-4-pyridone-5-carboxamide 2-Deoxyglucose 6-phosphate Gly-Asp Homocarnosine Glucaric acid o-Toluic acid XA0017 XC0016 Cholic acid Pantothenic acid Isobutyric acidSuccinicCDP-cholineAscorbic acid acid Fructose 6-phosphate N-Acetylasparagine Glutathione (GSSG)_divalent Butyric acid Sedoheptulose 7-phosphate S-Adenosylhomocysteine β-Ala HomoserineXC0089 Noradrenaline N6,N6,N6N-Acetylglucosamine-Trimethyllysine 6-phosphate Propionyl CoA_divalent XA0013 N-AcetylhistidineIsovalerylalanine-1 Ser-Glu6-Hydroxydopamine Pipecolic acid UDP-N-acetylgalactosamine-24-Pyridoxic acid Cysteinesulfinic acid N-AcetylleucineHomovanillic-1 acid N-Acetylgalactosamine UDP-N-acetylglucosamine-2 N-Acetylmannosamine Hypoxanthine 1-MethylnicotinamideTrimethylamine N-oxide 1-Methylhistidine4-Acetamidobutanoic acid N-Acetylglucosamine Pelargonic acid Thiamine3-Methylhistidine diphosphate UDP-glucose Triethanolamine myo-Inositol 2-phosphate Met UDPN--galactoseAcetylcysteineCTP 5,6,7,8-Tetrahydrobiopterin XA0012 Fructose 1,6-diphosphatethreo-β-Methylaspartic acid N-Acetyllysine Gluconolactone Gly-Gly 0 FAD_divalent PRPP Gly-Leu N8-Acetylspermidine Dihydroxyacetone phosphate Isovalerylalanine-2 Histaminecis-4-Hydroxyproline Terephthalic acid Spermine CDP N-Acetylleucine-2 N-AcetylserineADMA DecanoicThymidine acid XC0132 Arg-Glu DiethylaminomalonicMethionine acid sulfoxide N-Glycolylneuraminictrans-Glutaconic acid acid XA0035 Glycocholic acid N-Acetylaspartic acid 1-Aminocyclopropane-1-carboxylic acid-1 2'-Deoxycytidine cis-Aconitic acid Isoglutamic acid TyrosineHomoserinelactone methyl ester -1 ErgothioneineADP Propionic acid Imidazolelactic2- HydroxyvalericacidN-Acetylglutamic acid acid Kynurenine Asp SaccharopineNMN Glu PC2 N-Acetylglycine N-Acetylornithine Gluconic acid 2-Hydroxybutyric acidPhenaceturicCMP acid-N-acetylneuraminate Hydroxyproline Urea Choline S-AdenosylmethionineAscorbate 2-sulfate N,NCitrulline-Dimethylglycine UDP-glucuronic acid Creatinine γ-Glu-Cys O-Acetylhomoserine cAMP γ-Butyrobetaine O-Succinylhomoserine Octopamine 3'-Dephospho2-Aminoadipic CoA acid Octylamine XA0019Adenosine NAD+ N2-Acetylaminoadipic acid Pterin 1-MethylhistamineADP-ribose Glyceraldehyde 3-phosphate Cysteic acid Dopamine Ethyl glucuronide5-Aminovaleric acid N-Acetylglucosamine 1-phosphate XA0003 Glutaric acid 3-Indoxylsulfuric5'-Deoxy-5'-methylthioadenosine acid ATP Glucuronic acid-2 Anserine_divalent NADH 3-Hydroxybutyric acid 3-Aminoisobutyric acid N6-Methyllysine Thiamine phosphate N-Acetylneuraminic4-Amino acid-3-hydroxybutyric acid Galacturonic acid-2 Creatine 2-Hydroxyglutaric acid Ribose 5-phosphate5-Aminoimidazole5-Methyltetrahydrofolic-4-carboxamide ribotide acid CytidineArgininosuccinic acid Ethanolamine 2,6-Diaminopimelic acid N5-Ethylglutamine N-Acetyl-β-alanine Ascorbate N2--Acetylphenylalaninephosphate 1H-Imidazole-4-propionic acidFMN -0.05 3-Phenylpropionic acid Pyruvic acidXA0065 2-HydroxyXA0027γ-Glu-4--methylvaleric2-aminobutyric acid acid 3'-CMP XC0061 XC0137 4-DiethanolamineGuanidinobutyric acid N-Acetylmethionine 2'-CMP Theobromine5-Methoxyindoleacetic acid Quinic acid N-Acetylglucosylamine 3-Guanidinopropionic acid Isobutyrylcarnitine 7,8-Dihydrobiopterin GMP Ribulose 1,5-diphosphate 5-Oxoproline NADP+ Guanidoacetic acid AdipicNADPH_divalent acid Glucuronic acid-1 1-Pyrroline 5-carboxylic acid S-Nitrosoglutathione myo-Inositol 1-phosphateMucic acid Imidazole-4-acetic acidN-Ethylmaleimide_+H2O Betaine Phosphoenolpyruvic acid Galacturonic acid-1 myo-Inositol 3-phosphate CMP 2,3-Diphosphoglyceric acid O-Acetylcarnitine Adenosine 5'-phosphosulfate SerotoninPhosphocreatine SDMA Succinyl CoA_divalent Stachydrine CarboxymethyllysineMethylmalonyl CoA_divalent3',5'Erythrose-ADP 4-phosphate Betaine aldehyde_+H2O Malic acid Fumaric acid Guanosine CoA_divalent 3-Phosphoglyceric acid XA0033 Ectoine 3-Hydroxy-3-methylglutaric acid UMPOphthalmic acid Phosphorylcholine dCMP Ethanolamine phosphate

-0.1 Carnitine 1-Methyl-4-imidazoleacetic acid 2-Aminoisobutyric acid Suberic acid Citric acid 2-Aminobutyric acid Lauric acid Myristoleic acid

Adenylosuccinic acid Type2

-0.15 -0.15 -0.1 -0.05 0 0.05 0.1 0.15 PC1

Fig.2. Loading plot of principal component analysis. We define substances with strong association only with principal component 1 as Type 1 and substances with strong association only with principal component 2 as Type 2.

♂Non-irradiated

♂Irradiated

♀Non-irradiated

♀Irradiated

Fig.3. Relative amounts of metabolites illustrated on metabolic pathways (partial)