Doses Of Docosahexaenoic Acid Research Articles

Effect of high DHA and ARA fortification on lipid oxidation of infant formula powder

ABSTRACT Our objective was to assess the impact of fortified high-dose docosahexaenoic acid (DHA) and arachidonic acid (ARA) in infant formula powders (IFs) on stability. We compared and evaluated the stability of IFs containing high and low doses of DHA and ARA over a 6-month storage period at 37°C. While most formulations showed minimal changes in unsaturated fatty acid content of during storage, significant losses of DHA and ARA were observed in some high-dose formulations. Furthermore, this study noted consistent trends in the changes of vitamin C and vitamin E across both formula groups. Notably, the high-dose group formulas exhibited a higher presence of volatile organic compounds (VOCs) during storage, along with a more pronounced increase in thiobarbituric acid-reactive substance values (TBARS). In conclusion, the inclusion of high doses of DHA and ARA in IFs may compromise their oxidative stability.

The Eicosapentaenoic Acid and Docosahexaenoic Acid Content Reported in the Nutrition Panel of Fish Oil Supplements and Formula Milks Promoted for Young Children in Selected Markets

The aim of this study was to investigate the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) content found in fish oil supplements and formula milks for young children sold in Europe, the Middle East, and North Africa's local markets and online stores. To allow comparison of fish oil supplements and formula milks, information about the EPA and DHA content was collected from the nutrition panel of each product. A total of 79 fish oil supplements and 70 formula milks were studied. Out of the 32 fish oil supplements in local markets 12 products (37.5%) and of the 47 fish oil supplements in Amazon online stores 12 products (25.5%) contained more EPA than DHA. The highest DHA:EPA ratio for fish oil supplements was 4.6. The percentage of Recommended Dietary Intake for DHA, based on 250 mg daily, ranged from 3.2% to 558% for the fish oil supplements, and the range of 0.4% to 25.2% was found for the formula milks. Fish oil supplements provided high daily dose of DHA, which was significantly higher than that provided by the formula milks.

DHA-rich fish oil plays a protective role against experimental cerebral malaria by controlling inflammatory and mechanical events from infection

Every year, thousands of children, particularly those under 5 years old, die because of cerebral malaria (CM). Following conventional treatment, approximately 25% of surviving individuals have lifelong severe neurocognitive sequelae. Therefore, improved conventional therapies or effective alternative therapies that prevent the severe infection are crucial. Omega-3 (Ω-3) polyunsaturated fatty acids (PUFAs) are known to have antioxidative and anti-inflammatory effects and protect against diverse neurological disorders, including Alzheimer's and Parkinson's diseases. However, little is known regarding the effects of Ω-3 PUFAs against parasitic infections. In this study, C57BL/6 mice received supplemental treatment of a fish oil rich in the Ω-3 PUFA, docosahexaenoic acid (DHA), which was started 15 days prior to infection with Plasmodium berghei ANKA and was maintained until the end of the study. Animals treated with the highest doses of DHA, 3.0 and 6.0 g/kg body weight, had 60 and 80% chance of survival, respectively, while all nontreated mice died by the 7th day postinfection due to CM. Furthermore, the parasite load during the critical period for CM development (5th to 11th day postinfection) was controlled in treated mice. However, after this period all animals developed high levels of parasitemia until the 20th day of infection. DHA treatment also effectively reduced blood-brain barrier (BBB) damage and brain edema and completely prevented brain hemorrhage and vascular occlusion. A strong anti-inflammatory profile was observed in the brains of DHA-treated mice, as well as, an increased number of neutrophil and reduced number of CD8+ T leukocytes in the spleen. Thus, this is the first study to demonstrate that the prophylactic use of DHA-rich fish oil exerts protective effects against experimental CM, reducing the mechanical and immunological events caused by the P. berghei ANKA infection.

DHA, nutrient intake, and maternal characteristics as predictors of pregnancy outcomes in a randomised clinical trial of DHA supplementation

To investigate the relationships among docosahexaenoic acid (DHA) intake, nutrient intake, and maternal characteristics on pregnancy outcomes in a phase III randomised clinical trial designed to determine the effect of a DHA dose of 1000mg/day compared to 200mg/day on early preterm birth (<34 weeks gestation). A secondary aim of the phase III randomised trial was to explore the relationships among pregnancy outcomes (maternal red blood cell phospholipid (RBC-PL) DHA at delivery, preterm birth, gestational age at delivery, labor type, birth anthropometric measures, low birth weight, gestational diabetes, pre-eclampsia, and admission to a neonatal intensive care unit) in participants (n=1100). We used Bayesian multiple imputation and linear and logistic regression models to conduct an analysis of five general classes of predictor variables collected during the trial: a) DHA intake, b) nutrient intake from food and supplements, c) environmental exposure to tobacco and alcohol, d) maternal demographics, and e) maternal medical history. DHA supplementation lowered the risk of preterm birth and NICU admission, and increased gestation and birth weight as observed in the primary analysis. Higher maternal RBC-PL-DHA at delivery was associated with DHA supplementation and formal education of a bachelor's degree or higher. DHA supplementation and maternal age were associated with a higher risk of gestational diabetes. Total vitamin A intake was associated with longer gestation, while fructose and intake of the long chain omega-6 fatty acid, arachidonic acid, were associated with shorter gestation. Risk of preterm birth was associated with a history of low birth weight, preterm birth, pre-eclampsia, and NICU admission. Bayesian models provide a comprehensive approach to relationships among DHA intake, nutrient intake, maternal characteristics, and pregnancy outcomes. We observed previously unreported relationships between gestation duration and fructose, vitamin A, and arachidonic acid that could be the basis for future research. ClinicalTrials.gov (NCT02626299); December 10, 2015.

Baseline Findings of PreventE4: A Double-Blind Placebo Controlled Clinical Trial Testing High Dose DHA in APOE4 Carriers before the Onset of Dementia

IntroductionLower blood levels of the omega-3 polyunsaturated fatty acid docosahexaenoic acid (DHA) are correlated with worse cognitive functions, particularly among APOE ε4 carriers. Whether DHA supplementation in APOE ε4 carriers with limited DHA consumption and dementia risk factors can delay or slow down disease progression when started before the onset of clinical dementia is not known.MethodsPreventE4 is a double-blind, single site, randomized, placebo-controlled trial in cognitively unimpaired individuals with limited omega-3 consumption and dementia risk factors (n=368). Its objectives are to determine (1) whether carrying the APOE ε4 allele is associated with lower delivery of DHA to the brain; and (2) whether high dose DHA supplementation affects brain imaging biomarkers of AD and cognitive function.Results365 cognitively unimpaired individuals between 55 and 80 (mean age 66) were randomized to 2 grams of DHA per day or identically appearing placebo for a period of 2 years. Half the participants were asked to complete lumbar punctures at baseline and 6-month visits to obtain cerebrospinal fluid (CSF). The primary trial outcome measure is the change in CSF DHA to arachidonic acid ratio after 6 months of the intervention (n=181). Secondary trial outcomes include the change in functional and structural connectivity using resting state functional MRI at 24 months (n=365). Exploratory outcomes include the change in Repeatable Battery of the Assessment of Neuropsychological Status at 24 months (n=365).ConclusionsFindings from PreventE4 will clarify the brain delivery of DHA in individuals carrying the APOE ε4 allele with implications for dementia prevention strategies. Trial was registered as NCT03613844.

The Potential of DHA as Cancer Therapy Strategies: A Narrative Review of In Vitro Cytotoxicity Trials.

Docosahexaenoic acid (DHA), also known as omega-3 (n-3) polyunsaturated fatty acid (PUFA), is a natural compound that has demonstrated pharmacological activity against several malignant neoplasms. Available cancer treatments cause side effects, affect healthy cells, reduce the quality of life of patients and may cause resistance to antineoplastics. For these reasons, the search for new therapies is continuous. This narrative review aimed to compile information on in vitro experiments that study the cytotoxic effect of DHA or molecules derived from DHA in tumor and nontumor cells. This was performed to highlight the potential of DHA as a strategy for cancer therapy and to gather information, which will help researchers plan experimental designs and develop research to discover effective therapies against cancer. In addition, studies were presented that demonstrate the dose of DHA that can treat patients with cancer. Thus, a search was conducted for articles on the SCOPUS and Web of Science platforms, published until 2022, that analyzed the action of DHA against breast, lung, colorectal, prostate, stomach and liver cancers. Cytotoxic effects were observed in tumor and nontumor cell lines, and these results varied with the type of cell line studied, drug concentration, incubation time and treatment combination, i.e., with DHA alone, combined with other drugs and with molecules derived from DHA. In patients with cancer, in all analyzed studies, DHA intake was associated with eicosapentaenoic acid (EPA) and/or proteins to aid chemotherapy, and with this procedure, tumor reduction, chemotherapy tolerance and muscle mass gain were obtained. This work contributes to the community by demonstrating the possible applicability of DHA in the pharmaceutical area of oncological therapies.

Docosahexaenoic acid (DHA) intake estimated from a 7-question survey identifies pregnancies most likely to benefit from high-dose DHA supplementation.

Two randomized trials found women with low blood docosahexaenoic acid (DHA; an omega 3 fatty acid) had fewer early preterm births (<34 weeks gestation) if they were assigned to high dose DHA supplementation, however, there is currently no capacity for clinicians who care for pregnancies to obtain a blood assessment of DHA. Determining a way to identify women with low DHA intake whose risk could be lowered by high dose DHA supplementation is desired. To determine if assessing DHA intake can identify pregnancies that benefit from high dose DHA supplementation. This secondary analysis used birth data from 1310 pregnant women who completed a 7-question food frequency questionnaire (DHA-FFQ) at 16.8±2.5 weeks gestation that is validated to assess DHA status. They were then randomly assigned to a standard (200mg/day) or high dose (800 or 1000mg/day) DHA supplement for the remainder of pregnancy. Bayesian logistic regressions were fitted for early preterm birth and preterm birth as a function of DHA intake and assigned DHA dose. Participants who consumed less than 150mg/day DHA prior to 20 weeks' gestation (n=810/1310, 58.1%) had a lower Bayesian posterior probability (pp) of early preterm birth if they were assigned to high dose DHA supplementation (1.4% vs 3.9%, pp=0.99). The effect on preterm birth (<37 weeks) was also significant (11.3% vs 14.8%, pp=0.97). The DHA-FFQ can identify pregnancies that will benefit most from high dose DHA supplementation and reduce the risk of preterm birth. The DHA-FFQ is low burden to providers and patients and could be easily implemented in obstetrical practice.

Early and late preterm birth rates in participants adherent to randomly assigned high dose docosahexaenoic acid (DHA) supplementation in pregnancy

Intention-to-treat analyses do not address adherence. Per protocol analyses treat nonadherence as a protocol deviation and assess if the intervention is effective if followed. To determine the rate of early preterm birth (EPTB, <34 weeks gestation) and preterm birth (PTB, <37 weeks gestation) in participants who adhered to a randomly assigned docosahexaenoic acid (DHA) dose of 1000mg/day. Eleven hundred women with a singleton pregnancy were enrolled before 20-weeks' gestation, provided a capsule with 200mg/day DHA and randomly assigned to two additional capsules containing a placebo or 800mg of DHA. In the Bayesian Adaptive Design, new randomization schedules were determined at prespecified intervals. In each randomization, the group with the most EPTB was assigned fewer participants than the other group. Adherence was defined a priori as a postpartum red blood cell phospholipid DHA (RBC-PL-DHA) ≥5.5%.and post hoc as ≥8.0% RBC-PL-DHA, the latter after examination of postpartum RBC-PL-DHA. Bayesian mixture models were fitted for gestational age and dichotomized for EPTB and PTB as a function of baseline RBC-PL-DHA and dose-adherence. Bayesian hierarchical models were also fitted for EPTB by dose adherence and quartiles of baseline RBC-PL-DHA. Adherence to the high dose using both RBC-PL-DHA cut points resulted in less EPTB compared to 200mg [Bayesian posterior probability (pp)=0.93 and 0.92, respectively]. For participants in the two lowest quartiles of baseline DHA status, adherence to the higher dose resulted in lower EPTB (≥5.5% RBC-PL-DHA, quartiles 1 and 2, pp=0.95 and 0.96; ≥8% RBC-PL-DHA, quartiles 1 and 2, pp=0.94 and 0.95). Using the Bayesian model, EPTB was reduced by 65%, from 3.45% to 1.2%, using both cut points. Adherence also reduced PTB before 35, 36 and 37 weeks using both cut points (pp≥0.95). In general, performance of the nonadherent subgroup mirrored that of participants assigned to 200mg. Adherence to high dose DHA reduced EPTB and PTB. The largest effect of adherence on reducing EPTB was observed in women with low baseline DHA levels. gov (NCT02626299).

Docosahexaenoic Acid Ameliorates Contextual Fear Memory Deficits in the Tg2576 Alzheimer's Disease Mouse Model: Cellular and Molecular Correlates.

Docosahexaenoic acid (DHA), the most abundant polyunsaturated fatty acid in the brain, is essential for successful aging. In fact, epidemiological studies have demonstrated that increased intake of DHA might lower the risk for developing Alzheimer's disease (AD). These observations are supported by studies in animal models showing that DHA reduces synaptic pathology and memory deficits. Different mechanisms to explain these beneficial effects have been proposed; however, the molecular pathways involved are still unknown. In this study, to unravel the main underlying molecular mechanisms activated upon DHA treatment, the effect of a high dose of DHA on cognitive function and AD pathology was analyzed in aged Tg2576 mice and their wild-type littermates. Transcriptomic analysis of mice hippocampi using RNA sequencing was subsequently performed. Our results revealed that, through an amyloid-independent mechanism, DHA enhanced memory function and increased synapse formation only in the Tg2576 mice. Likewise, the IPA analysis demonstrated that essential neuronal functions related to synaptogenesis, neuritogenesis, the branching of neurites, the density of dendritic spines and the outgrowth of axons were upregulated upon-DHA treatment in Tg2576 mice. Our results suggest that memory function in APP mice is influenced by DHA intake; therefore, a high dose of daily DHA should be tested as a dietary supplement for AD dementia prevention.

The value of a two-armed Bayesian response adaptive randomization trial

ABSTRACT We investigate the value of a two-armed Bayesian response adaptive randomization (RAR) design to investigate early preterm birth rates of high versus low dose of docosahexaenoic acid during pregnancy. Unexpectedly, the COVID-19 pandemic forced recruitment to pause at 1100 participants rather than the planned 1355. The difference in power between number of participants at the pause and planned was 87% and 90% respectively. We decided to stop the study. This paper describes how the RAR was used to execute the study. The value of RAR in two-armed studies is quite high and their use in the future is promising.

Abstract 10905: Relationship of Eicosapentaenoic Acid and Docosahexaenoic Acid Dosage to Plaque Progression in Patients With Coronary Atherosclerosis Receiving Statin and Polyunsaturated Fatty Acid Therapy

Introduction: Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are omega-3 polyunsaturated fatty acids (PUFAs) extensively studied in atherosclerosis. Clinical endpoint and imaging studies have yielded convergent findings favoring pure EPA over mixed EPA/DHA formulations. The effects of EPA and DHA dose on coronary plaque dynamics, however, remain unknown, and could yield insights into the utility of these PUFAs in coronary artery disease. Methods: We considered longitudinal studies that compared coronary plaque change in patients receiving statin versus statin+PUFA therapy. Outcomes were % change in total and lipid plaque volume. Treatment effects were calculated as standardized mean difference (SMD), the mean % plaque volume change between statin and statin+PUFA cohorts divided by their pooled standard error. To isolate effects of EPA or DHA dose respectively, treatment arms were pooled by dosage of the PUFA of interest. To account for dependent samples, we fit hierarchical mixed effects models using robust variance estimation, assuming a correlation of 0.5 among treatment effects within multi-arm studies. Results: Among 555 screened articles, 11 studies met inclusion criteria. Among 1,070 patients, there were 14 treatment arms stratified by EPA dose, and 15 treatment arms stratified by DHA dose. Meta-regression revealed that EPA dose (g/day) correlated positively with treatment effect (β = 0.3 [0.1, 0.6], P = 0.02), while daily DHA dose correlated negatively with treatment effect (β = -0.4 [-0.7, -0.1], P = 0.01). We found no associations between EPA (β = 0.1 [-0.2, 0.4], P = 0.44) or DHA (β = -0.1 [-0.3, 0.2], P = 0.59) dose and treatment effects for lipid volume (Figure 1). Conclusions: Among cohorts receiving statin+PUFA therapy, we found a dose-dependent benefit of administered EPA on coronary plaque volume; the opposite was true for DHA. This adds to a growing body of evidence suggesting DHA may blunt beneficial effects of EPA in atherosclerosis.

Abstract 13395: Truth in Advertising? Health Claims and Contents of Fish Oil Supplements

Background: Fish oil supplement use continues to rise. Although prescription eicosapentaenoic acid (EPA) at 4 gm/day did lower cardiovascular risk, the vast majority of randomized trials have shown no benefit to fish oil dietary supplements. Information on health claims and contents of fish oil supplements is limited. Methods: Label information for on-market fish oil supplements was obtained from the National Institute of Health Dietary Supplement Label Database as of February 2022. Label text was analyzed to categorize health claims by type and organ system referenced. The total daily dose of EPA and docosahexaenoic acid (DHA) was evaluated for supplements made by 16 of the leading retailers and manufacturers of fish oil. Results: We evaluated 2,192 fish oil supplement labels, among which 1,647 (75.1%) made at least one health claim. Of these, only 22.0% (n=362) used from the FDA-approved qualified health claim for fish oil. The remaining 78.0% of labels made “structure &amp; function” claims (e.g., “promotes heart health”, “supports brain and vision function”, etc.) across a variety of organ systems (Figure). Heart-health related claims were the most common (n=1,390, 63.4% of labels). Across 16 different brands/manufacturers, 239 product labels were reviewed for dose information. Wide variation in the doses of EPA and DHA was observed (median EPA+DHA dose 600 mg, interquartile range 250-1100 mg). Only 9.2% of supplements reviewed contained at least 2 gm of EPA+DHA, the minimum recommended for triglyceride lowering, and none contained an EPA dose shown to prevent cardiovascular events (≥4 gm/day). Conclusions: Fish oil supplement labels commonly make vaguely worded claims that imply a range of health benefits, almost none of which are supported by randomized trial data. The doses of EPA and DHA available in commonly used fish oil supplements vary substantially, yet none contain levels shown to reduce cardiovascular risk.

Docosahexaenoic acid enrichment of tumor phospholipid membranes increases tumor necroptosis in mice bearing triple negative breast cancer patient-derived xenografts

Docosahexaenoic acid (DHA) reduces breast cancer tumor growth in preclinical models. To better understand how DHA amplifies the actions of docetaxel (TXT) chemotherapy, we examined the effects of two doses of dietary DHA on tumor size, membrane DHA content and necroptosis using a drug resistant triple negative breast cancer (TNBC) patient derived xenograft (PDX) model. Female NOD.Cb-PrkdcscidIl2rg mice bearing TNBC PDXs were randomized to one of three nutritionally complete diets (20% w/w fat): control (0% DHA), high DHA (3.8% HDHA), or low DHA (1.6% LDHA) with or without intraperitoneal injections of 5 mg/kg TXT, twice weekly for 6 weeks (n=8 per group). Tumors from mice fed either HDHA+TXT or LDHA+TXT were similar in size to each other, but were 36% and 32% smaller than tumors from mice fed control+TXT, respectively (P<.05). A dose effect of DHA incorporation was observed in plasma total phospholipids and in phosphatidylethanolamine and phosphatidylinositol. Both doses of DHA resulted in similarly increased necrotic tissue and decreased NFκB protein expression compared to control tumors, however only the HDHA+TXT had increased expression of necroptosis related proteins: RIPK1, RIPK3 and MLKL (P<.05). Increased MLKL was observed in the lipid raft portion of HDHA+TXT tumor extracts. This work confirms the efficacy of a combination therapy consisting of DHA supplementation and TXT chemotherapy using two doses of DHA as indicated by reduced tumor growth in a TNBC PDX model. Moreover, the results suggest that decreased growth may occur through increased DHA incorporation into tumor phospholipid membranes and necroptosis.

Harmonizing blood DHA levels in pregnancy studies: An interlaboratory investigation

Recent trials in pregnant women on the effects of supplemental DHA on early preterm birth (ePTB) risk have shown that there is a maternal blood docosahexaenoic acid (DHA) level below which risk for ePTB was increased and supplemental DHA was effective at reducing risk. However, DHA levels were expressed in different terms across these trials making cross study comparisons impossible. The purposes of this study were 1) to report interlaboratory conversion factors from study-specific metrics to a common metric, red blood cell (RBC) DHA measured by OmegaQuant Analytics (OQA), and 2) to translate reported pre- and post-treatment DHA levels from these trials into a RBC DHA for comparison. Data from five published and one unpublished study are included. Across these studies, the effects on RBC DHA levels after supplementation with 0, 200, 600, 800 and 1000 mg of DHA were (as a% change from baseline): 0 mg, no change; 200 mg, 15–20% increase; 600 mg, 55–60% increase; 800 mg, 13–65% increase; and 1000 mg, 51% increase. Standardization of fatty acid analysis and reporting and a target omega-3 or DHA level for identifying those for which higher dose DHA supplementation is indicated to prevent ePTB are needed for clinical use.

Entorhinal thickness: A marker of DHA supplementation efficacy?

AbstractBackgroundThe entorhinal cortex (ERC) area in the brain is affected early in AD, particularly among APOE4 carriers. We previously reported a positive association between plasma phospholipid docosahexaenoic acid (DHA) levels and the ERC thickness in the Aging Brain Program. We hypothesize that increases in plasma DHA after high‐dose DHA supplementation protects against thinning of the ERC when started before cognitive impairment including in APOE4 carriers, and that ERC thickness can be a useful marker for DHA supplementation efficacy.MethodWe examined the effect of DHA treatment and association of plasma DHA species on the changes in the ERC area in the DHA Brain Delivery Pilot Trial (NCT02541929). T1‐weighted MRI scans were collected at 3T and FreeSurfer software was used to segment the ERC thickness. Total DHA, DHA in phosphatidylcholine (PC), lysophosphatidylcholine (LPC), and phosphatidylethanolamine (PE) were measured in plasma together with Red blood cell (RBC) DHA levels before and after randomization to 2 grams of triglyceride DHA per day vs placebo over 6 months (n=28). Participants were all cognitively normal, consumed less than 200 mg of DHA per day, and were stratified by APOE4 genotype before randomization.ResultDHA treatment, adjusted for APOE4, was associated with a significant increase in ERC area thickness compared with placebo (mean group difference in ERC change (95% CI) 0.161 (0.003, 0.319), p=0.046). Among DHA subspecies measured, greater increase in plasma PC DHA levels after supplementation was significantly correlated with greater ERC area (mean ERC change per SD of PC DHA=0.103, p=0.011). The association persisted after adjusting for APOE4 status (mean ERC change per SD of PC DHA=0.109, p=0.007), indicating that the effect was independent of APOE4 status.ConclusionThese preliminary findings support a protective effect of high dose DHA in preserving the ERC thickness, including among APOE4 carriers. This finding supports early intervention, before the onset of cognitive impairment, with high dose DHA supplementation in individuals at risk of cognitive decline. PREVENTE4 trial (NCT03613844) is ongoing to help understand whether high dose DHA can prevent cognitive decline in individuals at increased risk of AD.HNY is supported by AA NIRG‐15‐361854, R01AG054434, R21AG056518, ADDF GC‐201711‐2014197.

SUPLEMENTASI DOCOSAHEXAENOIC ACID (DHA) SEBAGAI USAHA PREVENTIF IBU HAMIL DENGAN RISIKO PREEKLAMPSIA

Hypertension in pregnancy is one of the main cause of maternal death in the world. Hypertensive disorders in pregnancy, including preeclampsia, have a broad spectrum of conditions associated with maternal and fetal morbidity and mortality. Preeclampsia is a syndrome characterized by new onset of hypertension at ≥20 weeks of gestation, in pregnant women who were previously normotensive and accompanied by proteinuria. This paper aims to determine the effectiveness of docosahexaenoic acid (DHA) in pregnant women to prevent the risk of preeclampsia. This research method is to review the articles published in the last 10 years with the keywords preeclampsia, therapy, preventive, and docosahexaenoic acid (DHA). The data show that administration of docosahexaenoic acid (DHA) can reduce the risk of preeclampsia with minimal side effects for both maternal and the fetus. DHA on the other hand is still insignificant in preventing preeclampsia in high-risk pregnant women and prescribing the high dose of DHA is not recommended for pregnant women. DHA will be more effective in preventing preeclampsia if given in early pregnancy, especially in the first trimester of pregnancy

The Potential of Docosahexaenoic Acid (DHA) in SARS-CoV-2 Management: DHA Reduces ACE2 Levels in Endothelial Cells

ObjectivesAngiotensin-converting enzyme 2 (ACE2) is a transmembrane protein located on the surface of endothelial cells that promotes vasodilation by promoting the hydrolysis of angiotensin II. However, ACE2 also serves as the cellular receptor for SARS-CoV-2. Infection by SARS-CoV-2 can promote endothelial dysfunction which in turn is associated with greater infection severity. Long chain omega-3 fatty acids (n3 PUFA) like eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are reported to prevent endothelial dysfunction. Thus, we hypothesize that treatment with n3 PUFA may suppress the actions of SARS-CoV-2 on endothelial cells, potentially through endothelial ACE2. The objective of the study was therefore to investigate whether changes in ACE2 levels occur as part of the endothelial response to n3 PUFA treatment.MethodsMale fa/fa Zucker rats were randomised into 4 PUFA-based diet groups: α-linoleic acid (ALA), EPA, DHA, and linoleic acid (LA), for 8 weeks. EA.hy926 cells were cultured to sub-confluent and quiescent states and treated with various doses of DHA, EPA, and ALA for 8 or 24 h. ACE2 levels in tissues and cells were quantified by Western blotting.ResultsIn contrast to ALA and EPA, DHA treatment significantly reduced ACE2 levels in rat heart, aorta, and kidney but not lungs after 8 weeks of diet intervention. EPA only showed this reduction compared to ALA in kidney. Interestingly, when applied to human endothelial cells in culture, DHA decreased ACE2 levels of growing EA.hy926 cells even at relatively low doses, but no significant effect was observed in quiescent cells. EPA also had an effect, but only at an extremely high dose.ConclusionsDHA reduced ACE2 levels in key organs and human endothelial cells, which should produce beneficial effects by lowering the susceptibility of cells to SARS-CoV-2.Funding SourcesSt Boniface Hospital Foundation - Research Without Borders and University of Manitoba - GETS

Novel Mechanisms Related to DHA’s Atheroprotective Effects in Endothelial Cells: eNOS Activity Is Regulated by DHA via p38MAPK and MSK

Abstract Objectives Our laboratory previously reported that docosahexaenoic acid (DHA) activates p38 mitogen-activated protein kinase (MAPK) differently in growing and quiescent human endothelial cells, which represent the atherogenic and healthy states in vivo, respectively. Endothelial nitric oxide synthase (eNOS) activity differs between these two states. Since eNOS can be regulated by p38MAPK and mitogen-stimulated kinase (MSK) is a p38MAPK substrate involved in cell proliferation and inflammation, we hypothesized that DHA's atheroprotective actions require eNOS activation via the p38MAPK/MSK pathway. Thus, our objective was to investigate the role of p38MAPK/MSK in the eNOS response to DHA and determine whether the proposed pathway is growth state-sensitive. Methods EA.hy926 cells were cultured on Matrigel-coated plates to sub-confluent and quiescent states and treated with DHA ± SB202190 or SB747651A, inhibitors of p38MAPK and MSK, respectively. eNOS activation was quantified by Western blot detection of Ser1177 phosphorylation. Results eNOS activation by DHA in EA.hy926 cells was concentration-, time-, and growth state-dependent. p38MAPK inhibition suppressed eNOS activity in sub-confluent cells and increased eNOS activity in quiescent cells, while MSK inhibition suppressed eNOS activity in both growth states. eNOS activity remained suppressed with DHA treatment under MSK inhibition and showed no dose- or growth state-dependent effects. In contrast, when p38MAPK was inhibited, high dose DHA activated eNOS in sub-confluent cells, but dose-dependently decreased the elevated eNOS activity of quiescent cells. Conclusions eNOS activity in endothelial cells is modulated by DHA via p38MAPK and MSK. The growth state-dependent regulation of p38MAPK and eNOS by DHA provides novel insight into the molecular mechanisms of DHA's atheroprotective actions in relation to health status. Funding Sources Research Manitoba, St Boniface Hospital Foundation, University of Manitoba - GETS

Efficacy of Docosahexaenoic Acid for the Prevention of Necrotizing Enterocolitis in Preterm Infants: A Randomized Clinical Trial.

Necrotizing enterocolitis (NEC) is an inflammatory bowel disease and a leading cause of morbidity and mortality in preterm infants. In this study, a randomized double-blind parallel-group (1:1) trial was carried out in two neonatal intensive care units of two tertiary hospitals. Two hundred and twenty-five preterm newborns with an expected functional gastrointestinal tract were recruited and received an enteral dose of 75 mg of docosahexaenoic acid (DHA)/kg body weight or high-oleic sunflower oil daily for 14 days from the first enteral feed after birth. Confirmed NEC was evaluated with Bell’s scale from stage ≥ IIa. Two hundred and fourteen randomized infants were analyzed in terms of the intent-to-treat (DHA-group: n = 105; control-group: n = 109); data for two hundred infants were analysed per protocol. Confirmed NEC was lower in infants from the DHA-group compared with the control-group (0/100 vs. 7/100; p = 0.007), with RR = 0.93 (95% CI 0.881 to 0.981), risk difference = −7%, (95% CI −12.00 to −1.99), and number needed-to-treat = 15 (95% CI 8.3 to 50). Intent-to-treat analysis showed a lower level of treatment failure in the DHA-group compared with the control-group (6/105 (6%) vs. 16/109 (15%); p = 0.03, RR = 0.905, (95% CI 0.826 to 0.991)). The results after multivariate-regression analysis remained significant. Adverse events (apart from the incidence of NEC) were not different between groups. A daily dose of DHA for 14 days starting with the first enteral feed may prevent NEC in preterm infants.

Baseline red blood cell and breast milk DHA levels affect responses to standard dose of DHA in lactating women on a controlled feeding diet

BackgroundThe importance of providing the newborn infant with docosahexaenoic acid (DHA) from breast milk is well established. However, women in the United States, on average, have breast milk DHA levels of 0.20%, which is below the worldwide average (and proposed target) of >0.32%. Additionally, the relationship between maternal red blood cell (RBC) and breast milk DHA levels may provide insight into the sufficiency of DHA recommendations during lactation. Whether the standard recommendation of at least 200 mg/day of supplemental DHA during lactation is sufficient for most women to achieve a desirable RBC and breast milk DHA status is unknown. MethodsLactating women (n = 27) at about 5 weeks postpartum were enrolled in a 10–12 week controlled feeding study that included randomization to 480 or 930 mg choline/d (diet plus supplementation). As part of the intervention, all participants were required to consume a 200 mg/d of microalgal DHA. RBC and breast milk DHA levels were measured by capillary gas chromatography in an exploratory analysis. ResultsMedian RBC DHA was 5.0% (95% CI: 4.3, 5.5) at baseline and 5.1% (4.6, 5.4) after 10 weeks of supplementation (P = 0.6). DHA as a percent of breast milk fatty acids increased from 0.19% (0.18, 0.33) to 0.34% (0.27, 0.38) after supplementation (P<0.05). The proportion of women meeting the target RBC DHA level of >5% was unchanged (52% at baseline and week 10). The proportion of women achieving a breast milk DHA level of >0.32% approximately doubled from 30% to 56% (p = 0.06). Baseline RBC and breast milk DHA levels affected their responses to supplementation. Those with baseline RBC and breast milk DHA levels above the median (5% and 0.19%, respectively) experienced no change or a slight decrease in levels, while those below the median had a significant increase. Choline supplementation did not significantly influence final RBC or breast milk DHA levels. ConclusionsOn average, the standard prenatal DHA dose of 200 mg/d did not increase RBC DHA but did increase breastmilk DHA over 10 weeks in a cohort of lactating women in a controlled-feeding study. Baseline DHA levels in RBC and breast milk affected the response to DHA supplementation, with lower levels being associated with a greater increase and higher levels with no change or a slight decrease. Additional larger, dose-response DHA trials accounting for usual intakes and baseline DHA status are needed to determine how to best achieve target breast milk DHA levels and to identify additional modifiers of the variable breast milk DHA response to maternal DHA supplementation.