scholarly journals The fuel of respiration of rat kidney cortex

1969 ◽  
Vol 112 (2) ◽  
pp. 149-166 ◽  
Author(s):  
M. J. Weidemann ◽  
H. A. Krebs

1. In kidney-cortex slices from the well-fed rat, glucose (5mm) supplied 25–30% of the respiratory fuel; in the starved state, the corresponding value was 10%. These results are based on measurements of the net uptake of glucose and of the specific radioactivity of labelled carbon dioxide formed in the presence of [U−14C]-glucose. 2. Added acetoacetate (5mm) or butyrate (10mm) provided up to 80%, and added oleate (2mm) up to 50% of the fuel of respiration. The oxidation of endogenous substrates was suppressed correspondingly. 3. More [U−14C]oleate was removed by the tissue than could be oxidized by the amount of oxygen taken up; less than 25% of the oleate removed was converted into respiratory carbon dioxide and about two-thirds was incorporated into the tissue lipids. The rate of oleate incorporation into the neutral-lipid fraction was calculated to be equivalent to the rate of oxidation of endogenous fat, which provided the chief remaining fuel. 4. The contribution of endogenous substrates to the respiration (50%) in the presence of added oleate is taken to reflect either a high turnover rate of the endogenous neutral lipids (approx. half-life 2·5hr.) or a raised rate of lipolysis caused by the experimental conditions in vitro. 5. Added l-α-glycerophosphate (2·5mm) increased oleate incorporation into the neutral-lipid fraction by up to 40% (i.e. caused a net synthesis of triglyceride). 6. Lactate (2·5mm) added as sole substrate supplied 30% of the respiratory fuel, but with added oleate (2mm) lactate was converted quantitatively into glucose. Oleate stimulated the rate of gluconeogenesis from lactate by 45%. 7. The oxidation of both long-chain and short-chain even-numbered fatty acids was accompanied by ketone-body formation. Ketone-body synthesis from oleate, but not from butyrate, increased six- to seven-fold after 48hr. of starvation. The maximum rates of renal ketogenesis (80μmoles/hr./g. dry wt., with butyrate) were about 20% of the maximum rates observed in the liver (on a weight-for-weight basis) and accounted for, at most, 35% of the fatty acid removed. 8. dl-Carnitine (1·0mm) had no effect on the rates of uptake of acetate, butyrate or oleate or on the rate of radioactive carbon dioxide formation from [U−14C]oleate, but increased ketone-body formation from oleate by more than 100%. Ketone-body formation from butyrate was not increased. 9. There is evidence supporting the assumption that there are cells in which gluconeogenesis and ketogenesis occur together, characterized by equal labelling of [U−14C]oleate and the ketone bodies formed, and other cells that oxidize fat and do not form ketone bodies. 10. Inhibitory effects of unlabelled acetoacetate on the oxidation of [1−14C]butyrate and of unlabelled butyrate on [4−14C]acetoacetate oxidation show that fatty acids and ketone bodies compete as fuels on the basis of their relative concentrations. 11. The pathway of ketogenesis in renal cortex must differ from that of the liver, as β-hydroxy-β-methylglutaryl-CoA synthetase is virtually absent from the kidney. In contrast with the liver the kidney possesses 3-oxo acid CoA-transferase (EC 2.8.3.5), and the ready reversibility of this reaction and that of thiolase (EC 2.3.1.9) provide a mechanism for ketone-body formation from acetyl-CoA. This mechanism may apply to extrahepatic tissues generally, with the possible exception of the epithelium of the rumen and intestines.

1970 ◽  
Vol 119 (3) ◽  
pp. 525-533 ◽  
Author(s):  
H. A. Krebs ◽  
R. Hems

1. The formation of acetoacetate, β-hydroxybutyrate and glucose was measured in the isolated perfused rat liver after addition of fatty acids. 2. The rates of ketone-body formation from ten fatty acids were approximately equal and independent of chain length (90–132μmol/h per g), with the exception of pentanoate, which reacted at one-third of this rate. The [β-hydroxybutyrate]/[acetoacetate] ratio in the perfusion medium was increased by long-chain fatty acids. 3. Glucose was formed from all odd-numbered fatty acids tested. 4. The rate of ketone-body formation in the livers of rats kept on a high-fat diet was up to 50% higher than in the livers of rats starved for 48h. In the livers of fat-fed rats almost all the O2 consumed was accounted for by the formation of ketone bodies. 5. The ketone-body concentration in the blood of fat-fed rats rose to 4–5mm and the [β-hydroxybutyrate]/[acetoacetate] ratio rose to 11.5. 6. When the activity of the microsomal mixed-function oxidase system, which can bring about ω-oxidation of fatty acids, was induced by treatment of the rat with phenobarbitone, there was no change in the ketone-body production from fatty acids, nor was there a production of glucose from even-numbered fatty acids. The latter would be expected if ω-oxidation occurred. Thus ω-oxidation did not play a significant role in the metabolism of fatty acids. 7. Arachidonate was almost quantitatively converted into ketone bodies and yielded no glucose, demonstrating that gluconeogenesis from poly-unsaturated fatty acids with an even number of carbon atoms does not occur. 8. The rates of ketogenesis from unsaturated fatty acids (sorbate, undecylenate, crotonate, vinylacetate) were similar to those from the corresponding saturated fatty acids. 9. Addition of oleate together with shorter-chain fatty acids gave only a slightly higher rate of ketone-body formation than oleate alone. 10. Glucose, lactate, fructose, glycerol and other known antiketogenic substances strongly inhibited endogenous ketogenesis but had no effects on the rate of ketone-body formation in the presence of 2mm-oleate. Thus the concentrations of free fatty acids and of other oxidizable substances in the liver are key factors determining the rate of ketogenesis.


1998 ◽  
Vol 72 (2) ◽  
pp. 133-141 ◽  
Author(s):  
R.A. Holz ◽  
D.J. Wright ◽  
R.N. Perry

AbstractThe lipid composition of three batches of single generation cysts of Globodera rostochiensis, stored dry at 4°C for 1,7 and 13 years, comprised 81%, 74% and 53% neutral lipids, 14%, 18% and 27% non-acidic phospholipids and 5%, 8% and 20% free fatty acids, respectively. Lipids in eggs from two batches of G. pallida cysts, stored for 3 and 7 years, comprised 80% and 67% neutral lipids, 15% and 23% non-acidic phospholipids and 5% and 10% free fatty acids, respectively. All batches contained the same fatty acids which were dominated by C18:l, C20:l and C20:4. The fatty acid profiles of hatched J2 of G. rostochiensis from two batches, stored for 1 and 9 years, differed only in their free fatty acid fractions. Thus, while it is not possible to determine the age of cysts by their fatty acid profile, it may be possible to use the relative amounts of the main lipid classes as an indicator of age. Four batches of hatched J2 of G. pallida were investigated, with sample A hatched during the second week in potato root diffusate, B during week 3, C during week 4 and D during weeks 5 and 6 and stored for 3.5 days (on average) after hatching. Total lipid content was 27.2%, 31.5%, 18.5% and 6.3% of the dry weight for A, B, C and D, respectively. In the neutral lipid fraction of D an increase in C18:l and to a lesser extent C18:2 was observed. In the free fatty acid fraction of sample D, the percentages of C18:l, C18:2 and C18:3 were greater but the percentages of C20:3 and C20:4 were smaller compared with sample C. Fresh early hatched J2 of G. rostochiensis were compared with later hatched and stored (for 13 days on average) individuals for their lipid content and fatty acid composition. The lipid content was 26.1% and 11.4% in fresh and stored J2, respectively. Total lipid consisted of 77% and 70% neutral lipid, 18% and 26% non-acidic phospholipid and 6% and 4% free fatty acid in fresh and stored J2, respectively. In the neutral lipid fraction of stored J2 C18:l, C16:0 and C18:0 increased, whereas C20:4, C20:l and C20:3 decreased. Therefore, both neutral lipid and free fatty acid fractions showed changes in their fatty acid profiles after long delayed hatching and/or storage in both PCN species.


1998 ◽  
Vol 44 (11) ◽  
pp. 1051-1059 ◽  
Author(s):  
Anita Istokovics ◽  
Naoki Morita ◽  
Kazuo Izumi ◽  
Tamotsu Hoshino ◽  
Isao Yumoto ◽  
...  

The hyphae of the snow mold Microdochium nivale contained lipids in a yield of about 10% w/w of the dry matter of hyphae. The total lipid was fractionated into neutral and polar lipid fractions. In the neutral lipid fraction, triacylglylcerol was the sole major component. As minor components, ergosterol, diacylglycerol, free fatty acid, and fatty acyl ergosterol were identified. The polar lipid fraction contained phospholipids, glycolipids, and a lipid containing neither phosphorus nor sugar. Phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol, phosphatidylserine, and phosphatidic acid were identified as phospholipids. The polar lipid fraction included at least four kinds of glycolipids that have not been identified. A very unusual lipid in fungi, a betaine lipid, diacylglyceryltrimethylhomoserine, was identified by chemical and physicochemical analyses. The level of the neutral lipid fraction, which accounted for 60% of the total lipid in hyphae at the exponential phase, was significantly increased compared with that of the polar lipid fraction and constituted 80% of the total at the stationary phase. The neutral and polar lipids of Microdochium nivale contained 18:3 (9,12,15), 18:2 (9,12), 18:1 (9), and 16:0 as principal fatty acids. Among them, 18:2 (9,12) and 18:3 (9,12,15) were the major fatty acids of triacylglycerol, phosphatidylcholine, phosphatidylglycerol, and phosphatidylethanolamine, whereas in diacylglyceryltrimethylhomoserine, the major components were 16:0 and 18:3 (9,12,15).Key words: snow mold, phospholipids, betaine lipid, fatty acid, Microdochium nivale.


1988 ◽  
Vol 250 (3) ◽  
pp. 819-825 ◽  
Author(s):  
E P Brass ◽  
R A Beyerinck

Accumulation of propionate, or its metabolic product propionyl-CoA, can disrupt normal cellular metabolism. The present study examined the effects of propionate, or propionyl-CoA generated during the oxidation of odd-chain-length fatty acids, on hepatic oxidation of short- and medium-chain-length fatty acids. In isolated hepatocytes, ketone-body formation from odd-chain-length fatty acids was slow as compared with even-chain-length fatty acid substrates, and increased as the carbon chain length was increased from five to seven to nine. In contrast, rates of ketogenesis from butyrate, hexonoate and octanoate were all approximately equal. Propionate (10 mM) inhibited ketogenesis from butyrate, hexanoate and octanoate by 81%, 53% and 18% respectively. Addition of carnitine had no effect on ketogenesis from the even-chain-length fatty acids, but increased the rate of ketone-body formation from pentanoate (by 53%), heptanoate (by 28%) and from butyrate or hexanoate in the presence of propionate. The inhibitory effect of propionate could not be explained by shunting acetyl-CoA into the tricarboxylic acid cycle, as CO2 formation from butyrate was also decreased by propionate. Examination of the hepatocyte CoA pool during oxidation of butyrate demonstrated that addition of propionate decreased acetyl-CoA and CoA as propionyl-CoA accumulated. Addition of carnitine decreased propionyl-CoA by 50% (associated with production of propionylcarnitine) and increased acetyl-CoA and CoA. Similar changes in the CoA pool were seen during the oxidation of pentanoate. These results demonstrate that accumulation of propionyl-CoA results in inhibition of short-chain fatty acid oxidation. Carnitine can partially reverse this inhibition. Changes in the hepatocyte CoA pool are consistent with carnitine acting by generating propionylcarnitine, thereby decreasing propionyl-CoA and increasing availability of free CoA. The data provide further evidence of the potential cellular toxicity from organic acid accretion, and supports the concept that carnitine's interaction with the cellular CoA pool can have a beneficial effect on cellular metabolism and function under conditions of unusual organic acid accumulation.


1973 ◽  
Vol 134 (2) ◽  
pp. 499-506 ◽  
Author(s):  
Oliver E. Owen ◽  
Helene Markus ◽  
Stuart Sarshik ◽  
Maria Mozzoli

1. Concentrations of ketone bodies, free fatty acids and chloride in fed, 24–120h-starved and alloxan-diabetic rats were determined in plasma and striated muscle. Plasma glucose concentrations were also measured in these groups of animals. 2. Intracellular metabolite concentrations were calculated by using chloride as an endogenous marker of extracellular space. 3. The mean intracellular ketone-body concentrations (±s.e.m.) were 0.17±0.02, 0.76±0.11 and 2.82±0.50μmol/ml of water in fed, 48h-starved and alloxan-diabetic rats, respectively. Mean (intracellular water concentration)/(plasma water concentration) ratios were 0.47, 0.30 and 0.32 in fed, 48h-starved and alloxan-diabetic rats respectively. The relationship between ketone-body concentrations in the plasma and intracellular compartments appeared to follow an asymptotic pattern. 4. Only intracellular 3-hydroxybutyrate concentrations rose during starvation whereas concentrations of both 3-hydroxybutyrate and acetoacetate were elevated in the alloxan-diabetic state. 5. During starvation plasma glucose concentrations were lowest at 48h, and increased with further starvation. 6. There was no significant difference in the muscle intracellular free fatty acid concentrations of fed, starved and alloxan-diabetic rats. Mean free fatty acid intramuscular concentrations (±s.e.m.) were 0.81±0.08, 0.98±0.21 and 0.91±0.10μmol/ml in fed, 48h-starved and alloxan-diabetic states. 7. The intracellular ketosis of starvation and the stability of free fatty acid intracellular concentrations suggests that neither muscle membrane permeability nor concentrations of free fatty acids per se are major factors in limiting ketone-body oxidation in these states.


2011 ◽  
Vol 105 (6) ◽  
pp. 857-865 ◽  
Author(s):  
Kyriakoula Marinou ◽  
Martin Adiels ◽  
Leanne Hodson ◽  
Keith N. Frayn ◽  
Fredrik Karpe ◽  
...  

Before the menopause, women are relatively protected against CVD compared with men. The reasons for this sex difference are not completely understood, but hepatic fatty acid metabolism may play a role. The present study aimed to investigate the utilisation of plasma NEFA by the liver and to determine whether they are partitioned differently into ketone bodies and VLDL-TAG in healthy, lean young men and women. Volunteers were studied during a prolonged overnight fast (12–19 h) using an intravenous infusion of [U-13C]palmitate. After 12 h fasting, the women had a more advantageous metabolic profile with lower plasma glucose (P < 0·05) and TAG (P < 0·05) but higher plasma NEFA (P < 0·05) concentrations. Plasma 3-hydroxybutyrate (3-OHB) concentrations rose more in women than in men, and the transfer of13C from [U-13C]palmitate to plasma [13C]3-OHB reached a plateau 6–7 h after the start of the infusion in women but was still increasing at 6 h in men. This implies a slower 3-OHB production rate and/or dilution by other precursor pools in men. In women, the high isotopic enrichment of plasma 3-OHB suggested that systemic plasma fatty acids were the major source of 3-OHB production. However, in men, this was not observed during the course of the study (P < 0·01). There were no sex differences for the incorporation of13C into VLDL1- or VLDL2-TAG. The ability of young women to partition fatty acids towards ketone body production rather than VLDL-TAG may contribute to their more advantageous metabolic profile compared with young men.


1984 ◽  
Vol 32 (3) ◽  
pp. 474-476 ◽  
Author(s):  
Janis Cerbulis ◽  
Owen W. Parks ◽  
Ray H. Liu ◽  
Edwin G. Piotrowski ◽  
Harold M. Farrell

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