Biomedical subjects
J P Poisson
Publications and source records attributed to J P Poisson.
[Polyunsaturated fatty acid biogenesis is impaired in spontaneously hypertensive rat in relation to the pathogenesis of hypertension].
This study was designed to investigate polyunsaturated fatty acid (PUFA) biogenesis in spontaneously hypertensive rats (SHR) during the onset of hypertension. We measured liver desaturase activities, limiting enzymes of linoleic (LA) biosynthesis into arachidonic acid (AA). Fifteen male SHR and 15 male Wistar Kyoto (WKY) normotensive rats were killed at 7.9 or 13 week-old (5 per group). Systolic blood pressure was measured by plethysmography. Liver microsomes were obtained after ultracentrifugation and 5 mg of microsomal proteins incubated during 5 minutes at 37 degrees C with 0.04 mumoles of (1-14C) LA or (2-14C) dihomo-gamma-linolenic acid (DGLA). After fatty acid saponification and methylation, the bioconversion of (1-14C) LA into (1-14C) gamma-linolenic acid (GLA; delta 6 desaturation) and of (2-14C) DGLA into (2-14C) AA (delta 5 desaturation) was determined by measuring the radioactivity shared out between substrate and product of desaturation, after HPLC partition. Fatty acid composition of liver total lipids was calculated after GLC partition. In 7 week-old SHR, blood pressure was closed to that of WKY rats, higher in the 9 week-olds, when hypertension was settled in the 13 week-olds. Delta 6 and delta 5 desaturase activities were lower in SHR than in corresponding WKY. Those impaired desaturase activities increased with the age of rats: delta 6 desaturase activity was 43, 70 and 61% lower in the 7, 9 and 13 week-old SHR, respectively; delta 5 desaturase activity was 14, 48 and 53% lower in the same conditions. We observed a liver storage of total lipids in the 3 groups of SHR. The fatty acid composition of liver total lipids shows a lower proportion of AA and a higher proportion of LA in SHR than in WKY rats, confirming the depletion of the enzymatic systems studied. In summary, LA bioconversion into AA is decreased in liver microsomes of SHR, via the desaturase impairments. This phenomenon is concomitant with the pathogenesis of hypertension and related to the liver fatty acid composition. Such results explain partly the effects of PUFA rich diets administered to hypertensive rats, and clarify the interpretation of such effects.
Influence of spontaneous hypertension on n-3 delta-6-desaturase activity and fatty acid composition of rat hepatocytes.
The first rate limiting step in the conversion of alpha-linolenic acid is catalyzed by the delta-6-desaturase enzyme. The activity of such an enzyme was studied in order to investigate the n-3 Polyunsaturated Fatty Acid biogenesis during hypertension. Rat isolated hepatocyte n-3 delta-6-desaturase activity was higher in 1 month old Spontaneously Hypertensive Rats -- prehypertensive period-as compared to normotensive Wistar Kyoto rats, whereas there was no significant differences at 12 months -- hypertensive period-. Our data indicate no correlation between the directly measured enzyme activity and the changes in hepatocyte n-3 fatty acid compositions. The loss of hepatocyte n-3 delta-6-desaturase activity in the Spontaneously Hypertensive Rat may be a key factor in the evolution of hypertension related to aging through altering the eicosanoid balance.
Age-related depletion of linoleic acid desaturation in liver microsomes from young spontaneously hypertensive rats.
The present study was designed to investigate the microsomal interconversion of linoleic acid (LA) into arachidonic acid (AA) in young spontaneously hypertensive rats (SHR), in relation to the pathogenesis of hypertension. Our results show lower delta 6 and delta 5 desaturase activities (the limiting steps in the bioconversion of LA into AA) in young SHR, as compared to Wistar Kyoto normotensive rats. This impairment of desaturase activities is raised when the blood pressure increases and is related to the age of animals. The fatty acid composition of liver lipids shows a lower proportion of AA and a higher proportion of LA in SHR than in normotensive rats, confirming the depletion of the enzymatic system studied. Such a loss of desaturase activity may be under the control of hormones involved in the regulation of SHR blood pressure.
Age-related changes in linoleic acid bioconversion by isolated hepatocytes from spontaneously hypertensive and normotensive rats.
This study points out the hepatocyte interconversion of the linoleic acid family during hypertension. Hepatocyte delta 6 desaturase activity was higher in 1 month-old spontaneously hypertensive rats than in normotensive controls. A similar tendency was observed in 6 month-old SHR. delta 5 desaturase activity was higher only in 1 month-old spontaneously hypertensive rats as compared to controls. Desaturase activities were particularly high at the age of 6 months. The hepatocyte fatty acid composition showed an impairment of n-6 polyunsaturated fatty acid metabolism in spontaneously hypertensive animals. Changes were greater in the young prehypertensive rats than in adults. A storage of n-3 long chain fatty acids is remarkable in adult hypertensive rats, suggesting an alteration in peroxisomal oxidation. Such modifications may be related to the prostaglandin precursors availability to peripheral tissues such as kidney.
Incorporation of delta 6- and delta 5-desaturation fatty acids in liver microsomal lipid classes of obese Zucker rats fed n - 6 or n - 3 fatty acids.
The aim of this work was to study the effect of dietary n - 6 (as borage oil) and of n - 3 (as fish oil) fatty acids on the incorporation--in liver microsomal lipid classes--of fatty acids involved in delta 6- and delta 5-desaturations in obese Zucker rats compared with their lean littermates and with Wistar control rats. We observed that body and liver weights were decreased when obese Zucker rats were fed the fish oil diet. The major part of the radioactivity was recovered, in the obese Zucker rats, into the neutral lipids and especially into the triacylglycerols, while it was recovered into the phospholipid classes, especially into phosphatidylcholine, in the two other strains. Results show, in all phenotypes, an increased alpha-linolenic acid delta 6-desaturation in PL classes when the rats were fed the fish oil diet. However, a decreased linoleic acid delta 6- and delta 5-desaturation was observed in obese Zucker rats fed the fish oil diet. The fish oil diet favours the n - 3 fatty acid biosynthesis and incorporation into liver microsomal lipid classes to the prejudice of the n - 6 fatty acid series. The fatty acid incorporation is simultaneously regulated by the genetical phenotype and dietary fatty acids.
Age-related changes in antioxidant defence mechanisms and peroxidation in isolated hepatocytes from spontaneously hypertensive and normotensive rats.
The effects of age and hypertension on the antioxidant defence systems and the lipid peroxidation in rat isolated hepatocytes were studied. Four different age groups (1, 3, 6 and 12 months) were considered in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. Age-associated changes were observed on vitamin E status, glutathione (GSH) level, MDA formation and glutathione peroxidase (GSH-Px) activity in both strains. Maximal levels or activities of these parameters were found at 3 and 6 months, except for MDA which was low at 3 months. Then, a fall was observed at 12-month-old compared to 6-month values. In addition, GSH-Px activity was significantly lower in SHR than in WKY rats, except at the age of one month. The decrease of this enzyme activity could induce an increased cellular generation of radical species and lipid peroxidation, which might be link to hypertension.
Adrenic acid delta 4 desaturation and fatty acid composition in liver microsomes of spontaneously diabetic Wistar BB rats.
We examined the delta 4 (n-6) desaturation and the fatty acid composition of liver microsomes in the insulin-dependent spontaneously diabetic Wistar Bio-Breeding (BB) rat. The desaturation of adrenic acid to n-6 docosapentaenoic acid was decreased in the normo- and hyperglycemic diabetic rats. Insulin treatment with 1.0 IU. 100 g body weight-1 twice a day for 2 days restored the reduced activity during the hypoglycemic period. The pattern of responses was similar to that of linoleic acid delta 6 and dihomo-gamma-linolenic acid delta 5 desaturases, with a non-parallel relationship between the desaturation system and the glycemia. The microsomal fatty acid composition of BB rat liver reflected only partially to the delta 4 desaturation at different states of glycemia. Factors other than impaired desaturation system are involved in the fatty acid metabolism of spontaneously diabetic rats.
Psychosocial stress, catecholamines, and essential fatty acid metabolism in rats.
To examine the effects of psychosocial stress and the "stress hormone," epinephrine, on essential fatty acid metabolism in rats, two studies were conducted. In the first, the effects of four weeks of (i) social isolation and (ii) group housing (control) on liver microsomal delta 6 and delta 5 n-6 desaturase activity were studied in group-reared male normotensive (Wistar Kyoto) and spontaneously hypertensive (SHR) rats (n = 5/group). The second study examined the effects of acute ip epinephrine (0.0, 1.0, 2.0, and 4.0 mg/kg) 6 hr prior to and following an ig dose (4 g/kg) of safflower oil (rich in 18:2n-6, LA) on plasma and liver LA, 20:4n-6 (AA), and LA/AA ratios in adult essential fatty acid deficient Sprague-Dawley rats (n = 6/group). In the first experiment, isolation stress significantly inhibited the activity of delta 6 (P < 0.05) and delta 5 (P < 0.01) desaturase in the normotensive rats and of delta 5 desaturase in the SHR (P < 0.05). In the second study, epinephrine increased plasma and liver LA at doses 1.0 and 2.0 mg/kg in most of the fractions examined, and suppressed AA levels. The response of the LA/AA ratio to epinephrine varied between tissues and among lipid fractions, but increased this ratio at the moderate doses (2.0-4.0 mg/kg) of epinephrine in most cases. These data suggest that psychosocial stressors are capable of inhibiting the rate limiting steps of essential fatty acid metabolism and that this response is more pronounced in the SHR than in the Wistar Kyoto. They also suggest that epinephrine is capable of altering the in vivo metabolism of essential fatty acids in the rat.
Evidence that liver microsomes of human neonates desaturate essential fatty acids.
delta 6- and delta 5-Desaturation of essential fatty acids of n-6 and n-3 series are required for the biosynthesis of polyunsaturated fatty acids (PUFAs), which are precursors of eicosanoids and constituents of membrane phospholipids. This pathway could be of special importance during the perinatal period, when PUFAs accretion in the central nervous system is very active. However, experimental evidence of delta 6- and delta 5-desaturase activities in man is very scarce, and no data are available for newborns. We report the delta 6- and delta 5-desaturase activities detected in human liver microsomes from three neonates who died from associated malformations. Radiochemical assays of delta 6- and delta 5-desaturase activities performed with reverse phase HPLC analysis of the products in the n-6 series ranged from 4.8-13.6 to 3.2-16.4 pmol substrate converted.min-1.mg-1 microsomal proteins, respectively. In the n-3 series delta 6-desaturase activity ranged from 5.3 to 12.8 pmol.min-1.mg-1. The relationships between enzyme activities and substrate concentrations suggest excess substrate inhibition for n-6 and not for n-3 fatty acids. These results demonstrate significant delta 6- and delta 5-desaturase activities in human liver of neonates, but this activity was lower than previously reported in adult humans and in mammals, especially rodents.
Effect of salt-loading and spontaneous hypertension on in vitro metabolism of [1-14C]linoleic and [2-14C]dihomo-gamma-linolenic acids.
The present study compared the effect of spontaneous hypertension and salt-loading on in vitro metabolism of 18:2n-6 (linoleic acid) and 20:3n-6 (dihomo-gamma-linolenic acid). Ten weanling spontaneously hypertensive rats (SHR) and 10 normotensive Wistar-Kyoto rats (WKY) maintained on a rodent lab chow were given tap water with (n = 5) or without (n = 5) addition of 1% NaCl for 4 weeks. Thereafter, animals were killed and liver microsomes were prepared. Aliquots of microsomes suspended in the phosphate-sucrose buffer containing MgCl2, ATP, CoA, and NADPH were incubated with 0.3 microCi of [1-14C]-18:2n-6 or [2-14C]-20:3n-6 at 37 degrees C for 15 min. The activity of delta 6- and delta 5-desaturases, and the distribution of radioactivity in different lipid fractions and in phospholipid fatty acids were determined. Results showed that both spontaneous hypertension and salt-loading suppressed the desaturation of radiolabeled 18:2n-6 and of 20:3n-6. Incubation of microsomes with [1-14C]-18:2n-6 resulted in 29% of radioactivity being associated with phospholipid fraction, of which 3% was associated with 18:3n-6. Incubation with radiolabeled 20:3n-6 resulted in 30% of the radioactivity being incorporated into phospholipids, of which 50% was associated with 20:4n-6 (arachidonic acid). Salt-loading suppressed the incorporation of radiolabeled fatty acids into phospholipids, more so in SHR than in WKY. Thus, salt-loading not only suppressed the desaturation of 18:2n-6 and 20:3n-6, but also interfered with the acylation of n-6 fatty acids into the phospholipid fraction.
Age-related changes in delta 6 and delta 5 desaturase activities in rat liver microsomes.
Age-related changes in delta 6 desaturation of [1-14C]alpha-linolenic acid and [1-14C]linoleic acid and in delta 5 desaturation of [2-14C]dihomo-gamma-linolenic acid were studied in liver microsomes from Wistar male rats at various ages ranging from 1.5 to 24 mon. Desaturase activities were expressed both as specific activity of liver microsomes and as the capacity of whole liver to desaturate by taking into account the total amount of liver microsomal protein. delta 6 Desaturation of alpha-linolenic acid increased from 1.5 to 3 mon and then decreased linearly up to 24 mon to reach the same desaturation capacity of liver measured at 1.5 mon. The capacity of liver to desaturate linoleic acid increased up to 6 mon and then remained constant, whereas microsomal specific activity was equal at 1.5 and 24 mon of age. The capacity of liver to convert dihomo-gamma-linolenic acid to arachidonic acid by delta 5 desaturation decreased markedly from 1.5 to 3 mon. It then increased to reach, at 24 mon, the same level as that observed at 1.5 mon. Age-related changes in the fatty acid composition of liver microsomal phospholipids at the seven time points studied and of erythrocyte lipids at 1.5 and 24 mon were consistent with the variations in desaturation capacity of liver. In particular, arachidonic acid content in old rats was slightly higher than in young rats whereas contents in linoleic and docosahexaenoic acids varied little throughout the life span.(ABSTRACT TRUNCATED AT 250 WORDS)
Altered desaturase activities and fatty acid composition in liver microsomes of spontaneously diabetic Wistar BB rat.
We examined the activities of delta 9, delta 6 and delta 5 desaturases and fatty acid composition of liver microsomes in the insulin-dependent spontaneously diabetic adult female Wistar Bio-Breeding (BB) rat. The diabetic BB rats were subcutaneously injected with different doses of protamine zinc insulin in order to be killed in hyper-, normo- or hypo-glycemic states. Desaturase activities, which are partially inhibited by spontaneous diabetes during the normo- and hyper-glycemic periods, were similarly affected by the various insulin treatment; delta 9 desaturase activity being more depressed than the desaturase activities of either delta 6 of delta 5. Insulin treatment with 10 I.U./kg body weight twice a day for 2 days was able to restore the delta 9, delta 6 and delta 5 desaturase activities to control levels during the hypoglycemic period. The microsomal fatty acid composition of BB rats liver was not consistent with the desaturase activities, particularly delta 9 desaturase activity, during the different states of glycemia, indicating that they are not closely linked in a direct cause-effect relationship.
Evidence for insulin dependent hepatic microsomal gamma-linolenic acid chain elongation in spontaneously diabetic Wistar BB rats.
We studied hepatic microsomal gamma-linolenoyl-CoA elongation and fatty acid composition of liver microsomes in spontaneously diabetic Wistar BB rats. The liver microsomal gamma-linolenoyl-CoA elongation was decreased in diabetic Wistar BB rats during both normo- and hyperglycemic periods and restored during the hypoglycemic period following insulin treatment. These results are in agreement with our previously reported data on linoleic acid delta 6 and delta 5 desaturations and support the non-parallel relationship between the chain elongation system and the glycemia. The fatty acid composition of BB rat liver microsomes was only partially consistent with the gamma-linolenoyl-CoA elongation activity at the different periods of glycemia, probably because factors other than elongation impairments were involved in the evolution of fatty acid composition.
Effect of sodium loading (3% NaCl) on arachidonic acid biosynthesis in rat liver microsomes.
Sodium loading increases arachidonic acid (AA) metabolism by way of the prostaglandins(PGs) from series 2. Its effect on AA biosynthesis remains unknown. The purpose of the present study was to investigate the influence of sodium loading on the fatty acid composition of liver and liver microsomes, and the liver microsomal delta-6 and delta-5 desaturations of linoleic acid (LA) into AA. We found a decrease of LA and dihomo-gamma-linolenic acid (DGLA) levels in liver total lipids of Wistar rats receiving hypernatriuretic drinking water (NaCl 3%) for 60 days. At the same time AA increased. DGLA decreased and AA increased in liver microsomal total lipids. 1(14) C-LA delta-6 desaturase and 2(14) C-DGLA delta-5 desaturase activities increased in liver microsomes. These results show that, in addition to its influence on the regulation of glomerular filtration, sodium loading is involved in the regulation of liver AA biosynthesis.
Depletion of delta 9 desaturase (EC 1.14.99.5) enzyme activity in growing rat during dietary protein restriction.
The effects of protein restriction on delta 9 desaturase (EC 1.14.99.5) activity were studied in growing rats. A control group was fed on a balanced diet (200 g casein/kg; BD) for 28 d. The experimental group was fed on the low-protein diet (20 g casein/kg; LP) for 26 d, then refed the balanced diet (BD-R) for 2 d. Rats were born to and suckled from normally fed dams. The enzyme activity was measured after 2 and 14 d of LP, and 26 d of LP plus 2 d of BD-R, by incubations in vitro of hepatic microsomal pellets with [1-14C]steric acid. The results indicated a decreased delta 9 desaturase activity after 2 and 14 d of LP of -33 and -43% respectively. Refeeding for 2 d was sufficient to super-repair this activity (+66%). The fatty acid composition of total liver lipids and microsomal phosphatidylethanolamines (PE) and phosphatidylcholines (PC) were also investigated; 18:0 decreased in total liver lipids at 14 d of LP, when 18:1n-9 increased. Stearic acid (18:0) increased in PC at 2 d of LP and in PE at 14 d of LP; oleic acid (18:1n-9) did not change. Therefore, it is concluded that a defect occurred in the bioconversion of 18:0 into 18:1n-9 by delta 9 desaturation during protein depletion. As oleic acid is accumulated in total liver lipids during LP, we speculate that this is due to a decreased oxidation or transport of this fatty acid.
Elongation and desaturation of arachidonic and eicosapentaenoic acids in rat liver. Effect of clofibrate feeding.
The fatty acid elongation-desaturation ability of 5,8,11,14-eicosatetraenoic (20:4(n-6)) and 5,8,11,14,17-eicosapentaenoic (20:5(n-3)) acids was determined in both liver microsomal and light mitochondrial (rich in peroxisomes) fractions of untreated and clofibrate treated rats. The elongation and the subsequent desaturation steps were performed in the corresponding favorable media. 20:5(n-3) elongation was about 2-times more extensive than that of 20:4(n-6). Clofibrate feeding for 10 days resulted in a marked decrease in the elongation rate with the two substrates, while the delta 4 desaturation rate was increased. There were small differences in the elongation rate between the microsomal and light mitochondrial fractions, however, the relative delta 4 desaturation rate was higher in the light mitochondrial fraction than microsomes.