The effect of dietary α-linolenic acid levels on regulation of omega-3 lipid synthesis in rat
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1 The effect of dietry α-linolenic cid levels on regultion of omeg-3 lipid synthesis in rt Wei-Chun Tu School of Agriculture Food nd Wine The University of Adelide
2 Conversion of PUFA to LCPUFA PUFA LCPUFA Omeg 3 Omeg 6 8:3n-3 ALA 6D D :n-3 EPA :n-3 DPA :n-3 6D :6n-3 :6n-3 DHA desturtion elongtion desturtion elongtion elongtion desturtion β oxidtion 8:n-6 LA 6D D :n-6 AA :n-6 DTA :n-6 6D :n-6 :n-6
3 Low PUFA LA:ALA : Elongse :3n-6 :n-3 8:3n-6 8:n-3 Dietry ALA increse :n-6 :6n-3 6 Desturse β oxidtion 8:n-6 (LA) 8:3n-3 (ALA) :n-6 :n-3 :n-6 :6n-3 (DHA) Elongse :n-6 :n-3 (DPA) Desturse :n-6 (AA) :n-3 (EPA) Elongse
4 Incresed DHA levels in the ody of humns nd nimls hve een chieved y incresing the mount of dietry ALA (Goyens, P. L. et l, Am J Clin Nutr 6; Aruckle, L. D. et l, J Nutr 99) or lowering the LA to ALA rtio in the diets (Blnk, C. M. et l, J Lipid Res ; Mkrides, M. et l, Am J Clin Nutr )
5 Heptic D-6 desturse ctivity is induced y diets low in essentil ftty cids nd suppressed y diets rich in vegetle or fish oils (Melin, T., nd Nilsson, A., Prostglndins Leukot Essent Ftty Acids 997; Peluffo, R. O. et l, Biochim. Biophys. Act 976) High in dietry n-3 nd n-6 PUFA sttus decrese DD/D6D mrna levels nd ctivities (Cho, H. P. et l, J Bio Chem 999) Dietry PUFA Fds, Elovl gene expression Desturse, elongse ctivities Fds: delt desturse gene Fds: delt 6 desturse gene Elovl: elongse gene Elovl: elongse gene
6 Omeg-3 rich Vegetle oils Chep, Stle, Sustinle With n optimised ALA level Aims Ftty cid synthetic pthwy LA (8:n-6) ALA Elongse Desturse 6 Desturse β oxidtion :n-6 DHA AA EPA Elongse :n-6 DPA Elongse High level n-3 LCPUFA ccumultion in tissues To evlute whether Incresed n-3 LCPUFA conversion cn e chieved y optimizing dietry LA:ALA rtio & PUFA levels Desturses nd elongses gene cn e ffected y dietry PUFA resluted in n-3 LCPUFA conversion
7 Dietry Design A rnge of ALA levels were tested while the level of other competing PUFA (LA) were held constnt t %en Three week-old Hooded Wistr Diet x Mc oil x Soy oil x Exp. x Exp. x Exp. 3 x Exp. x Exp. LA %e ALA %e weeks All diets % ft, % protein lood plsm liver Ftty Acid Extrction RNA Extrction & qrt-pcr Anlysis
8 LA nd ALA in plsm PL % of ftty cids in rt plsm PL 6 8 LA:ALA Rtio 3 Soy oil LA:ALA Rtio Mc oil LA ALA Diet LA %e ALA %e LA:ALA Exp.. 7. Exp... Exp. 3.. Exp..9.6 Exp..9. Mc oil..3 Soy oil.
9 Ftty Acid Composition plsm n-3 LCPUFA (% totl ftty cid) 8 6 c c LA: %e d c d d ALA (% e) e plsm AA (% totl ftty cid) EPA DPA DHA AA Mc oil LA%en. ALA%en.3 Plsm PL Soy oil LA%en ALA%en. % of plsm PL n-3 LCPUFA LA:ALA Rtio % of plsm PL AA EPA DPA DHA A A % of ftty cids in rt plsm PL EPA DPA DHA AA Soy oil Mc oil
10 Ftty Acid Composition LA: %e liver n-3 LCPUFA (% totl ftty cid) d c c c c e d c e d c ALA (% e) liver AA (% totl ftty icd) EPA DPA DHA AA Mc oil LA%en. ALA%en.3 Liver PL Soy oil LA%en ALA%en. % of liver PL n-3 LCPUFA 6 8 LA:ALA Rtio EPA % of liver PL AA DPA DHA A A % of ftty cids in rt liver PL 3 EPA DPA DHA AA Soy diet Mc diet
11 Summry Ftty cid in plsm reflected dietry PUFA levels A significnt increse in EPA mtched y n decrese of plsm AA in plsm nd liver Low dietry rtio of LA to ALA nd high energy level of ALA resulted in high n-3 LCPUFAs in plsm nd liver EPA nd DPA levels in plsm nd liver incresed in dose dependent mnner
12 Wht hppen to the gene expression of key enzymes in the n-3 LCPUFA conversion pthwy?
13 Heptic gene expression level Ftty cid level in liver PL % of ftty cids in rt liver PL 8 6 EPA DPA DHA Soy diet Mc diet
14 Gene expression level Delt- desturse Fds Delt-6 desturse Fds Fold chnge 3 Fold chnge 3 3 ALA%e 3 ALA%e Elongse Elovl Elongse Elovl Fold chnge 3 Fold chnge 3 3 ALA%e 3 ALA%e
15 Gene expression level SREBP-c PPARα Fold chnge 3 Fold chnge 3 3 ALA%e 3 ALA%e Sterol regultory element inding proteins (SREBP) re memrne ound trnscription fctors. Liver express predominntly SREBP-c tht ctivtes genes of ftty cid synthesis. Peroxisome prolifertor ctivted receptors (PPAR) re single polypeptides nucler receptors. Liver express PPARαtht ctivtes trnscription of genes involve in in ftty cid βoxidtion.
16 Conclusions Dietry PUFA with lower LA:ALA rtio nd higher ALA %en results in significnt incorportions of DPA nd EPA plsm nd liver phospholipids Enzyme competition effects my e stronger thn gene expression on driving n-3 LCPUFA conversion
17 Acknowledgements Dr. Roert Gison, Dr. Jo Zhou, Dvid Apps, El Zielinski nd memers of Nutrition nd Functionl Food group Dr. Michel Jmes, Dr. Reecc Cook-Johnson Dr. Beverly Mühlhäusler
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