Measurement of acetyl-CoA carboxylase activity in isolated hepatocytes

Bijleveld, Caspaar; Geelen, M.J.H.

doi:10.1016/0005-2760(87)90231-1

Cited by 68 publications

(57 citation statements)

References 32 publications

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“…The total incubation mixture for the assay contained 78 mM Hepes (pH 7.5), 125 mM mannitol, 2.5 mM 2-mercaptoethanol, 3.1 mM EDTA, 4.0 mM citrate, 5.6mM MgClz, 2.0 mM ATP, 20 mM KHCOB, 0.93% (w/v) bovine serum albumin (charcoal treated and dialyzed), 0.44 mM dithioerythritol, 0.5 mM NADPH, 0.062 mM butyryl-CoA, 0.062 mM [1-i4C]acetyl-CoA (4 Ci/mol), and 3.2 mU fatty acid synthase. The rest of this procedure was performed exactly as described in (17).…”

Section: Methodsmentioning

confidence: 99%

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Effects of ethanol feeding on hepatic lipid synthesis

Tijburg

Maquedano

Bijleveld

et al. 1988

Archives of Biochemistry and Biophysics

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Rats were fed a high-fat, liquid diet containing either 36% of total calories as ethanol or an isocaloric amount of sucrose, for a period up to 35 days. At different time intervals we measured the effects of ethanol administration on the activities of a number of key enzymes involved in hepatic lipid synthesis. At the start of the experimental period the activities of acetyl-CoA carboxylase and fatty acid synthase, measured in liver homogenates, increased in the control as well as in the ethanol-fed group. After 35 days these enzyme activities were still elevated but there were no significant differences between the two groups. In hepatocytes isolated from controls as well as from ethanol-fed rats, short-term incubations with ethanol induced an increase in the rate of fatty acid synthesis and in the activities of acetyl-CoA carboxylase and fatty acid synthase. However, no alterations in the regulation of these enzymes by short-term modulators of lipogenesis were apparent in hepatocytes isolated from alcohol-treated animals. The results do not indicate a major role for the enzymes of de ncruo fatty acid synthesis in the development of the alcoholic fatty liver. The amount of liver triacylglycerols increased in ethanol-fed rats during the entire treatment period, whereas the hepatic levels of phosphatidylcholine and phosphatidylethanolamine were not affected by ethanol ingestion. Ethanol administration for less than 2 weeks increased the activities of phosphatidate phosphohydrolase, diacylglycerol acyltransferase, and microsomal phosphocholine cytidylyltransferase, whereas the cytosolic activity of phosphocholine cytidylyltransferase was slightly decreased. Upon prolonged ethanol administration the activities of these enzymes were slowly restored to control values after 35 days, suggesting development of some kind of adaptation. It is interesting that, although the activities of phosphatidate phosphohydrolase and diacylglycerol acyltransferase were restored to the levels found in the control rats, this effect was not accompanied by a stabilization or decrease of the concentration of hepatic triacylglycerols. 0 1988 Academic press, inc.

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Section: Methodsmentioning

confidence: 99%

“…Acetyl-CoA carboxylase activity in isolated hepatocytes was determined by a coupled assay in digitonin-permeabilized cells exactly as described before (17).…”

Section: Methodsmentioning

confidence: 99%

Effects of ethanol feeding on hepatic lipid synthesis

Tijburg

Maquedano

Bijleveld

et al. 1988

Archives of Biochemistry and Biophysics

Self Cite

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“…Liver slices were homogenized (25% homogenate wt/vol) in a buffer containing 10 mM HEPES (pH 7.5) and 0.25 mM sucrose and then centrifuged for 5 minutes at 12,000 g. The reactions were initiated by adding 0.5 mg proteins from the centrifuged (12,000 g) supernatant into a prewarmed buffer consisting of 200 mM HEPES, pH 6.8, 250 mM sucrose, 1 mM NADPH, 0.88 mM DTT, 2 mM EDTA, 2% BSA, 0.39 mM malonyl-CoA, and 0.124 mM [1-14 C]-acetyl-CoA (Amersham Biosciences). Newly synthesized 14 C-labeled fatty acids were extracted and quantified as described (30).…”

Section: Animals and Dietsmentioning

confidence: 99%

The fat-derived hormone adiponectin alleviates alcoholic and nonalcoholic fatty liver diseases in mice

et al. 2003

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Adiponectin has recently been shown to be a promising candidate for the treatment of obesity-associated metabolic syndromes. Replenishment of recombinant adiponectin in mice can decrease hyperglycemia, reverse insulin resistance, and cause sustained weight loss without affecting food intake. Here we report its potential roles in alcoholic and nonalcoholic fatty liver diseases in mice. Circulating concentrations of adiponectin decreased significantly following chronic consumption of highfat ethanol-containing food. Delivery of recombinant adiponectin into these mice dramatically alleviated hepatomegaly and steatosis (fatty liver) and also significantly attenuated inflammation and the elevated levels of serum alanine aminotransferase. These therapeutic effects resulted partly from the ability of adiponectin to increase carnitine palmitoyltransferase I activity and enhance hepatic fatty acid oxidation, while it decreased the activities of two key enzymes involved in fatty acid synthesis, including acetyl-CoA carboxylase and fatty acid synthase. Furthermore, adiponectin treatment could suppress the hepatic production of TNF-α and plasma concentrations of this proinflammatory cytokine. Adiponectin was also effective in ameliorating hepatomegaly, steatosis, and alanine aminotransferase abnormality associated with nonalcoholic obese, ob/ob mice. These results demonstrate a novel mechanism of adiponectin action and suggest a potential clinical application of adiponectin and its agonists in the treatment of liver diseases.

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“…ACC activity was determined by spectrophotometry as described previously with modification (Bijleveld et al, 1987). The assay was conducted in a mixture, containing 50 mM Tris-HCl (pH 7.6), 10 mM potassium citrate, 10 mM Mgcl2 , 3.75 mM Glutamate, 0.75 mg bovine serum albumin in 1 ml and 0.125 mM acetyl coa, 20 units of pyruvate kinase and 20 units of lactate dehydrogenase, adjusted with distilled water to 1 ml.…”

Section: Acetyl-coa Carboxylase Activity Assaymentioning

confidence: 99%

Effect of Urtica dioica leaf extract on activities of nucleoside diphosphate kinase and acetyl coenzyme, a carboxylase, in normal and hyperglycemic rats

Qujeq¹

2011

Afr. J. Pharm. Pharmacol.

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Urtica dioica has been shown to have capability of reducing blood glucose through enhancing insulin secretion. The aim of this study was to assess in vivo and in vitro effects of aqueous and alcoholic extracts of U. dioica leaves on activities of Acetyl coenzyme A carboxylase (ACC), Nucleoside diphosphate kinase (NDPK) and insulin level and serum glucose concentration. The experimental animals were randomly distributed among 6 groups of 7 or 8 rats each. The treatment groups received 150 mg/kg of alloxan. Then rats were fed alcoholic and aqueous extracts of U. dioica at 50 mg/kg/day doses for a period of two weeks. Subsequently glucose, insulin levels, and ACC, NDPK activity were measured. Our studies showed significantly lower levels of glucose in the group of rats that were treated with ethanol extract of U. dioica leaves as compared with the control group (279.6 ± 24.3 vs. 393.6 ± 24.9 mg/dl), (p<0.01). Present results showed significantly elevated levels of insulin in the group of rats treated with ethanol extract of U. dioica leaves as compared with the control group (7.6 ± 0.3 vs. 3.6 ± 0.2, µg/l), (p<0.01). Also, results showed significantly elevated activity of ACC and NDPK in the group of rats treated with ethanol and aqueous extract of U. dioica leaves as compared with the control group. The results of the present study indicated that alcoholic extract of U. dioica leaves was found to reduce glucose level and increase insulin secretion, ACC and NDPK activity in the alloxan diabetic animals.

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Measurement of acetyl-CoA carboxylase activity in isolated hepatocytes

Cited by 68 publications

References 32 publications

Effects of ethanol feeding on hepatic lipid synthesis

Effects of ethanol feeding on hepatic lipid synthesis

The fat-derived hormone adiponectin alleviates alcoholic and nonalcoholic fatty liver diseases in mice

Effect of Urtica dioica leaf extract on activities of nucleoside diphosphate kinase and acetyl coenzyme, a carboxylase, in normal and hyperglycemic rats

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