Endogenous Metabolite; Na+/K+ ATPase

The most prevalent unsaturated fertilizer (FA) in human fat cells and other tissues is oleic acid. Oleic acid increases β-oxidation-mediated AMPK activation-mediated cell proliferation and migration of highly metastatic cells. Oleic acid inhibits the growth and death of low-proliferation tumors, including the breast cancer MCF-7 cell line, as well as SGC7901 [1].

Dietary oleic acid intake improved running endurance with the changes of muscle fiber type shares in mice. This study elucidated a novel functionality of oleic acid in skeletal muscle fiber types. Further studies are required to elucidate the underlying mechanisms. Our findings have the potential to contribute to the field of health and sports science through nutritional approaches, such as the development of supplements aimed at improving muscle function.[3]

Oil Red O Staining and Triglyceride Assay[2]
The lipid droplets were stained by Oil red O. A stock solution was prepared in 2-propanol (0.3%), and a working solution was freshly prepared by diluting the stock solution with water (3∶2). After fixation, the cells were washed twice in PBS and stained with Oil red O and hematoxylin for 15 min and 1 min, respectively. The cells were washed with PBS, and images were obtained under a light contrast microscope. The intracellular triglycerides were assayed using a triglyceride assay kit (GPO-POD) according to the manufacturer’s recommended protocol.

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Determination of Oxygen Consumption[2]
Five million cells were resuspended in 1 ml of fresh warm medium pre-equilibrated with 21% oxygen and placed in a sealed respiration chamber equipped with a thermostat control, micro stirring device and Clark-type oxygen electrode disc. The oxygen content in the cell suspension medium was constantly monitored for 10 min, and the oxygen consumption rate was recorded.

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ATP Assays[2]
The intracellular level of ATP was measured with an ATPlite assay kit. The cells were incubated in a serum-free medium with or without 400 µM of OA for 48 h, washed with PBS three times, lysed in ATP extraction buffer and centrifuged in 4°C. ATP was measured by luminometric methods using commercially available luciferin/luciferase reagents on a luminometer (TD-20/20) according to the manufacturer’s instructions. The data were normalised to total protein.

Cell Viability Assay[2]
The cells in 48-well plates at a density of 10,000 cells per well were treated with different stressors. The cell viability was measured using the 3-[4,5-dimethylthiazol-2-yl]-2,5-dephenyl tetrazolium bromide (MTT) assay, according to the manufacturer’s protocol.

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BrdU Incorporation Assay[2]
The cell proliferation was determined by measuring the BrdU incorporation using a BrdU incorporation assay, according to the manufacturer’s instructions. Briefly, 5,000 cells/well seeded in a 96-well plate were pulse-labelled for 2 h with 10-uM BrdU. The cells were incubated for 30 min with a diluted, peroxidase-conjugated anti-BrdU antibody. The absorbance values were measured at 450 nm using an ELISA reader.

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Migration Assay[2]
The cell migration assays were performed using Transwell chambers (8-µm pore size polycarbonate membrane). A total of 50 K cells were plated into the insert in 200 µl serum-free medium containing BSA or 400 µM BSA-bound oleic acid and allowed to migrate from the upper compartment to the lower compartment toward a 15% FBS gradient for the indicated time period. After the migration, the non-migratory cells on the upper membrane surface were removed by scrubbing, and the membrane was fixed in buffered 4% paraformaldehyde and stained with 0.1% crystal violet at room temperature. The migrated cells were then enumerated. The migration values were expressed as the average number of migrated cells per microscopic field over six fields per assay from three independent membrane experiments.

All animal experiments were conducted in strict accordance with the Guidelines for Proper Conduct of Animal Experiments published by the Science Council of Japan and with the approval of the Animal Care and Use Committee of Kitasato University (approval no. 20-007). And all methods were performed in accordance with guidelines and regulations at Kitasato University, as well as complying with ARRIVE guidelines for the reporting of animal experiments. Eight-week-old male C57BL/6JJcl mice were purchased from CLEA Japan, Inc. The experimental design is illustrated in Fig. 5. The mice were housed in plastic cages in an animal room at 22 ± 2 °C and 50 ± 10% humidity under an artificial lighting system of 12-h light/12-h dark cycle. They were acclimated to the environment for one week. Following the acclimatization period, the mice were fed a CE-2 diet supplemented with 10% (w/w) palmitic acid (control diet) or oleic acid for 4 weeks. Palmitic acid was chosen as control fatty acid because it is a representative saturated fatty acid commonly found in soybean oil and olive oil, which has been used in previous studies. Although we considered using linoleic acid as a control, we decided against it because unsaturated fatty acids have ligand activity for PPARs. In our previous study, we reported the results of an 8-week feeding test, and in a preliminary study we confirmed similar changes in skeletal muscle characteristics even with a 4-week period. Therefore, we chose a shorter period of 4 weeks for this study. Nutritional and fatty acid compositions of experimental diets are shown in Table 2 and 3, respectively. Running endurance tests and serum biochemical analyses were performed on the last day of week 3 and the first day of week 4, respectively. After a 4-week feeding period, the mice were euthanized by cervical dislocation under inhalation anesthesia with isoflurane. The adipose tissues, liver, and skeletal muscles (soleus, EDL, and gastrocnemius muscles) were harvested and weighed immediately. The growth performance and tissue weight are shown in Table 1. All tissues were stored at –80 °C until further analysis [3].

Absorption, Distribution and Excretion
Fatty acid uptake by different tissues may be mediated via passive diffusion to facilitated diffusion or a combination of both. Fatty acids taken up by tissues are then stored in the form of triglycerides or oxidized. Oleic acid was shown to penetrate rat skin. Following oral administration of Brucea javanica oil emulsion in rats, the time of oleic acid to reach peak plasma concentration was approximately 15.6 hours.
Following oral administration of trace amounts of oleic acid, less than 10% of total oleic acid was found to be eliminated via fecal excretion.
Radio-labelled oleic acid was detected in the heart, liver, lung, spleen, kidney, muscle, intestine, adrenal, blood, and lymph, and adipose, mucosal, and dental tissues. Oleic acid is primarily transported via the lymphatic system.
No pharmacokinetic data available.
Radioactivity has been traced to the heart, liver, lung, spleen, kidney, muscle, intestine, adrenal, blood, and lymph, and adipose, mucosal, and dental tissues after administration of radioactive oleic, palmitic, and stearic acids.
Simultaneous ingestion of trace amounts of 14C-triolein (10 uCi) and 3H-oleic acid (20 uCi) in 42 g of carrier fat by patients with normal fecal fat excretion resulted in estimated fecal excretion of less than 10% of both substances. Gastrointestinal transit times for 14C-triolein, 3H-oleic acid, and a nonabsorbable marker, CrCl3, did not differ significantly.
Oleic Acid has been reported to penetrate the skin of rats. On histological examination, fluorescence from absorbed oleic acid was found in epidermal cell layers of skin removed from treated rats within 10 min of its application. The path of penetration was suggested to be via the hair follicles. Only minute amounts of oleic acid were visualized in the blood vessels throughout the experiment. Skin permeability was shown to increase with the lipophilic nature of a compound.
METABOLISM OF TRITIATED OLEIC ACID WAS STUDIED IN RATS DURING 600 DAYS. DURING FIRST 4 DAYS, HALF ACTIVITY IS FIXED TO WATER & HALF IS STORED IN ADIPOSE TISSUE WHICH IT LEAVES QUICKLY, THEN MORE SLOWLY WITH T/2 OF ABOUT 200 DAYS.
For more Absorption, Distribution and Excretion (Complete) data for OLEIC ACID (10 total), please visit the HSDB record page.

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Metabolism / Metabolites
Like most fatty acids, oleic acid may undergo oxidation via beta-oxidation and tricarboxylic acid cycle pathways of catabolism, where an additional isomerization reaction is required for the complete catabolism of oleic acid. Via a series of elongation and desaturation steps, oleic acid may be converted into longer chain eicosatrienoic and nervonic acid.
Proposed mechanisms for fatty acid uptake by different tissues range from passive diffusion to facilitated diffusion or a combination of both. Fatty acids taken up by the tissues can either be stored in the form of triglycerides (98% of which occurs in adipose tissue depots) or they can be oxidized for energy via the beta-oxidation and tricarboxylic acid cycle pathways of catabolism.
The beta-oxidation of fatty acids occurs in most vertebrate tissues (except the brain) using an enzyme complex for the series of oxidation and hydration reactions resulting in the cleavage of acetate groups as acetyl-CoA (coenzyme A). An additional isomerization reaction is required for the complete catabolism of Oleic Acid. Alternate oxidation pathways can be found in the liver (omega-oxidation) and in the brain (alpha-oxidation).
Fatty acid biosynthesis from acetyl-CoA takes place primarily in the liver, adipose tissue, and mammary glands of higher animals. Successive reduction and dehydration reactions yield saturated fatty acids up to a 16-carbon chain length. Stearic Acid is synthesized by the condensation of palmitoyl-CoA and acetyl-CoA in the mitochondria, and Oleic Acid is formed via a mono-oxygenase system in the endoplasmic reticulum.
The normal metabolic pathway of palmitic and stearic acids in mammals produces oleic acid. Oleic acid, on a series of elongation and desaturation steps, may be converted into longer chain eicosatrienoic and nervonic acid.
Weanling rats were fed diets containing rapeseed, canbra or ground nut oils for 8 or 60 days. They received simultaneously (14)C erucate and (3)H2 oleate by iv application. Animals were killed 2 or 19 hr after injection, lungs were removed and the distribution of (14)C and (3)H radioactivities was determined in pulmonary lipid fractions and in fatty acids of phospholipids and neutral lipids. More (14)C than (3)H radioactivity was recovered in lung lipids 3 and 19 hr after admin of labelled fatty acids. (14)C and (3)H radioactivity in the phospholipid fraction was larger than in the triglyceride fraction, the inverse was observed after 19 hr. The main part of (14)C radioactivity was present in the monounsaturated fatty acids, in decr order: 18:1, 24:1, 16:1 and 20:1. Erucic acid was slightly esterified in phospholipids.
Oleic acid has known human metabolites that include 18-Hydroxyoleic acid and 17-Hydroxyoleic acid.

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Biological Half-Life
No pharmacokinetic data available.
METABOLISM OF TRITIATED OLEIC ACID WAS STUDIED IN RATS DURING 600 DAYS. /ELIMINATION OCCURED/ SLOWLY WITH T/2 OF ABOUT 200 DAYS.

Protein Binding
As with other fatty acids originating from adipose tissue stores, oleic acid may bind to serum albumin or remain unesterified in the blood.

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Interactions
LOW CONCN OF ... OLEIC ACID ... CAUSED CONSIDERABLE INCREASE IN THE INTESTINAL ABSORPTION OF AMORPHOUS & POLYMORPHIC CHLORAMPHENICOL IN THE CAT.
LIVER TRITIATED-LABELED OLEATE SHOWED ACCUMULATION OF NEWLY SYNTHESIZED TRIGLYCERIDES WITHOUT ANY EFFECT ON PHOSPHOLIPIDS, AFTER SINGLE INJECTION OF CEROUS CHLORIDE IN RATS. /TRITIATED-LABELED OLEATE/
ALTERATIONS IN PHOSPHOLIPID COMPOSITION IN BRAIN & HEART OCCURS IN RESPONSE TO ETHANOL IN THOSE STRAINS OF MICE THAT SHOW RAPID TOLERANCE TO ETHANOL. AN INCREASE IN LIVER PHOSPHOLIPIDS CONTAINING OLEIC ACID WERE FOUND IN ALL STRAINS.
AFTER 16 MIN OF BRAIN ISCHEMIA IN RATS, BRAIN OLEATE INCREASED 2.5-FOLD. POSTISCHEMIA THERAPY WITH THIOPENTAL ACCELERATED THE RATE OF FALL OF BRAIN OLEATE. /OLEATE/
For more Interactions (Complete) data for OLEIC ACID (11 total), please visit the HSDB record page.

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Non-Human Toxicity Values
LD50 Rat oral 74 g/kg
LD50 Rat iv 2.4 mg/kg
LD50 Mouse iv 230 mg/kg
LD50 Guinea pig dermal >3000 mg/kg

[1]. Jack-Hays MG, et al. Activation of Na+/K(+)-ATPase by fatty acids, acylglycerols, and related amphiphiles: structure-activity relationship. Biochim Biophys Acta. 1996 Feb 21;1279(1):43-8.
[2]. Li S, et al. High metastaticgastric and breast cancer cells consume oleic acid in an AMPK dependent manner. PLoS One. 2014 May 13;9(5):e97330.
[3]. Sci Rep. 2024 Jan 8;14(1):755. doi: 10.1038/s41598-023-50464-y.

Therapeutic Uses
/EXPTL THER/ Ten Japanese boys with childhood adrenoleukodystrophy (ALD), one adult patient with adrenomyeloneuropathy (AMN), and two presymptomatic ALD boys were treated with dietary erucic acid (C22:1) for more than 12 months; except in a case of childhood ALD patient who died 7 months after beginning erucic acid therapy. During erucic acid therapy, the serum levels of very long-chain fatty acid (VLCFA) (C24:0/C22:0) decreased within 1-2 months in all patients, and these levels in four of the patients decreased to the normal range. Neurological examination and MRI findings in all 10 of the childhood ALD patients showed progression of the disease while they were receiving the dietary therapy. However, the mean interval between the onset of awkward gait and a vegetative state in diet-treated patients was significantly longer than that in the untreated patients. One AMN patient showed slight improvement of spastic gait and lessened pain in the lower limbs due to spasticity. The two presymptomatic ALD boys remained intact on clinical examination and on MRI findings for 38 and 23 months, respectively, after starting the diet.
/EXPL THER/ An open 2 yr trial of oleic and erucic acids (Lorenzos oil) included 14 men with adrenomyeloneuropathy, 5 symptomatic heterozygous women and 5 boys with preclinical adrenomyeloneuropathy. No evidence of a clinically relevant benefit from dietary treatment in patients with adrenomyeloneuropathy (accumulation of very-long-chain fatty acids) could be found. /Lorenzos oil/

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Drug Warnings
40 male and 6 female patients with adrenoleukodystrophy received Lorenzos oil (20% erucic acid and 80% oleic acid). In 19 of these patients the platelet count decr significantly. In 6 patients with thrombocytopenia, platelet counts became normal within 2 to 3 mo after erucic acid was omitted from the diet. Observations suggested that strategies for the dietary management of adrenoleukodystrophy requiring the admin of large amt of erucic acid may be associated with thrombocytopenia and that the erucic acid component of Lorenzos oil is the cause of the thrombocytopenia. Patients treated with erucic acid should be followed closely with determinations of the platelet count. /Lorenzos oil: 20% erucic acid and 80% oleic acid/
15 men with adrenoleukodystrophy and 3 symptomatic heterozygous women were admin oleic and erucic acids (Lorenzos oil). Asymptomatic thrombocytopenia developed in 5 patients (platelet counts ranged between 37000 and 84000 per cu mm) but was reversed within 2 to 3 wk after erucic acid was omitted. In addition, long-term treatment with Lorenzos oil (for 24 to 43 mo) was associated with lymphocytopenia in these 5 patients. The observations suggested that the long-term treatment of adrenoleukodystrophy with Lorenzos oil can induce severe lymphocytopenia with immunosuppression and recurrent infections. /Lorenzos oil/

C18H34O2

282.468

282.255

C, 76.54; H, 12.13; O, 11.33

112-80-1

Sodium oleate;143-19-1;Oleic acid-13C;82005-44-5;Oleic acid-d2;5711-29-5;Glycerol Monoleate;25496-72-4;Oleic acid-13C18;287100-82-7;Oleic acid-d17;223487-44-3;Oleic acid-13C-1;2483735-58-4;Oleic acid-d9;2687960-84-3

445639

Colorless to light yellow liquid

0.9±0.1 g/cm3

360.0±0.0 °C at 760 mmHg

13-14 °C(lit.)

270.1±14.4 °C

0.0±1.7 mmHg at 25°C

1.467

Endogenous Metabolite

7.7

1

2

15

20

234

0

O([H])C(C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])/C(/[H])=C(/[H])\C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H])=O

ZQPPMHVWECSIRJ-KTKRTIGZSA-N

InChI=1S/C18H34O2/c1-2-3-4-5-6-7-8-9-10-11-12-13-14-15-16-17-18(19)20/h9-10H,2-8,11-17H2,1H3,(H,19,20)/b10-9-

9-Octadecenoic acid (9Z)-

D 100 Oleic acid9-cis-Octadecenoic acid9Z-Octadecenoic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Ethanol : ~100 mg/mL (~354.03 mM)
0.1 M NaOH : ~100 mg/mL (~354.03 mM)
DMSO : ≥ 62.5 mg/mL (~221.27 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.85 mM) (saturation unknown) in 10% EtOH + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear EtOH stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (8.85 mM) (saturation unknown) in 10% EtOH + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear EtOH stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (8.85 mM) (saturation unknown) in 10% EtOH + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear EtOH stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: ≥ 2.08 mg/mL (7.36 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 5: ≥ 2.08 mg/mL (7.36 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 6: ≥ 2.08 mg/mL (7.36 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)