scholarly journals Linoleic Acid Reduces Apoptosis via NF-κB During the in Vitro Development of Porcine Embryos.

2020 ◽  
Author(s):  
Chang-Kyu Lee ◽  
Dong-Kyung Lee ◽  
Kwang-Hwan Choi ◽  
Jong-Nam Oh ◽  
Seung-Hun Kim ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Transcription Factor ◽  
Linoleic Acid ◽  
Protein Expression ◽  
Embryo Development ◽  
Metabolic Energy ◽  
Mrna Levels ◽  
Apoptosis Pathway ◽  
Apoptotic Gene

Abstract Background: Recent studies suggest that endogenous and exogenous free fatty acids play many important roles in mammalian oocyte and preimplantation embryo development. Among these fatty acids, linoleic acid has been reported to affect the apoptosis pathway via nuclear transcription factor-kappa B. The transcription factor NF-κB is a key modulator of apoptosis in a variety of cell types, but to date, this specific function of NF-κB has not been demonstrated in porcine preimplantation embryos. To examine the effect of linoleic acid on parthenogenetic pig embryos produced in vitro, we treated these embryos with linoleic acid at various concentrations to examine the developmental rate, NF‐κB expression, IL-6 expression and apoptosis-related gene mRNA levels. Results: Linoleic acid had a positive effect on embryo development and was not toxic at a certain concentration, but toxicity was observed at higher concentrations. Furthermore, it was confirmed that the concentration of NF‐κB increased, unlike that of IL-6, as the concentration of linoleic acid increased, and the concentration of NF‐κB was found to increase even at the concentration of linoleic acid at which embryo development decreased. We found that pro-apoptotic gene expression was downregulated in the linoleic acid-treated group. It was also found that BCL-XL, an anti-apoptotic gene, was not affected by linoleic acid, which appears to be an effect of IL-6. In contrast, MCL-1, an anti-apoptotic gene known to be unaffected by IL-6, was found to be increased at the mRNA level in linoleic acid-treated pig embryos. Furthermore, based on both apoptosis and immunocytochemistry staining, as the concentration of NF-kB increased, the nuclear translocation of C-JUN, which is also related to apoptosis, gradually increased, which was dependent on the linoleic acid concentration. It was confirmed that NF-κB is an important factor in the development of porcine embryos by confirming that treatment with a very low concentration of ammonium pyrrolidinedithiocarbamate (APDC, inhibitor of NF-κB) affected NF-κB protein expression, IL-6 protein expression and blastocyst production. Conclusion: These datas could suggest that porcine embryos can use exogenous linoleic acid as a metabolic energy source via NF-κB. The data also demonstrate the important role of NF-kB in porcine early embryo development.

81 Linoleic acid required for reduction of apoptosis through nuclear transcription factor-kappa B during pig embryo development

2020 ◽  
Vol 32 (2) ◽  
pp. 167
Author(s):  
D. Lee ◽  
K. Choi ◽  
J. Oh ◽  
S. Kim ◽  
M. Lee ◽  
...  
Keyword(s):  
Gene Expression ◽  
Fatty Acids ◽  
Transcription Factor ◽  
Linoleic Acid ◽  
Embryo Development ◽  
Mrna Level ◽  
Metabolic Energy ◽  
Apoptotic Gene ◽  
Nuclear Transcription Factor ◽  
Apoptotic Genes

Recent studies suggest that endogenous and exogenous free fatty acids play various important roles in mammalian oocyte and pre-implantation embryo development. Among fatty acids, linoleic acid (LA) has been reported to affect the apoptosis pathway via nuclear transcription factor-kappa B (NF-κB). The transcription factor NF-κB is a key modulator of apoptosis in a variety of cell types, but to date, this specific function of NF-κB has not been demonstrated in porcine pre-implantation embryos. To examine the effect of linoleic acid on invitro-produced parthenogenetic pig embryos, we treated LA by concentration (0, 10, 25, 50, and 100 µM) to identify developmental rate, NF-κB expression, and mRNA level of apoptotic-related genes. In addition, the mechanism was confirmed by examining the protein and mRNA expression of NF-kb and c-jun by immunostaining and quantitative PCR at the blastocyst stage. Linoleic acid had a positive effect on embryo development without toxicity at a certain concentration (25 µM), but toxicity was confirmed at higher (50-100μM) concentrations. Furthermore, it was confirmed that the concentration of NF-κB increased as the treatment concentration of LA increased, which was found to increase even at the concentration at which embryo development decreased. Previous studies have shown that the NF-κB pathway is involved in regulating anti- and pro-apoptotic gene expression. We also investigated the effects of LA on anti- (Bcl-xL, Mcl-1) and pro- (BAX1, TP53, Caspase3) apoptotic genes and NF-κB activation-related genes (RelA, JNK1, JNK2, IL-6) in porcine embryos. We have found that down-regulation of pro-apoptotic gene expression occurs in the LA-treated group. It was also found that Bcl-xL, one of the anti-apoptotic genes, was not affected by LA, which appears to be an effect of IL-6. In contrast, Mcl-1, an anti-apoptotic gene known not to be affected by IL-6, was found to have increased expression mRNA level in LA-treated pig embryos. Furthermore, through double-staining of apoptosis and immunocytochemistry, as the concentration of NF-kB level increases, the nuclear translocation of c-jun, the protein of which was also related with apoptosis, increased gradually depending on the LA concentration. These data could support that porcine embryo can use exogenous LA as a metabolic energy source. The data also demonstrate the important role of NF-kB in porcine early embryo development. Support was provided by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through the Development of High Value-Added Food Technology program funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA, 118042-03-1-HD020).

scholarly journals Eicosapentaenoic acid and 3,10 dithia stearic acid inhibit the desaturation of trans-vaccenic acid into cis-9, trans-11-conjugated linoleic acid through different pathways in Caco-2 and T84 cells

2006 ◽  
Vol 95 (4) ◽  
pp. 688-695 ◽  
Author(s):  
Renaville Bénédicte ◽  
Anne Mullen ◽  
Fiona Moloney ◽  
Yvan Larondelle ◽  
Yves-Jacques Schneider ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Linoleic Acid ◽  
Stearic Acid ◽  
Conjugated Linoleic Acid ◽  
Vaccenic Acid ◽  
Regulatory Element ◽  
Mrna Levels ◽  
T84 Cells ◽  
Key Enzyme

Stearoyl-CoA desaturase (SCD) is a key enzyme that determines the composition and metabolic fate of ingested fatty acids, in particular the conversion of trans-vaccenic acid (TVA) to conjugated linoleic acid (CLA). The present study addressed the hypothesis that intestinal TVA absorption and biotransformation into CLA can be modulated by EPA and 3,10-dithia stearic acid (DSA) via altered SCD mRNA levels and desaturation indices (cis-9, trans-11-CLA:TVA and oleic acid:stearic acid ratios) in Caco-2 and T84 cells, two well-established in vitro models of the human intestinal epithelium. The study determined the effect of acute (3h with 0·3mm-EPA or 0·3mm-DSA) and acute-on-chronic (1 week with 0·03mm-EPA or -DSA, followed by respectively, 0·3mm-EPA or -DSA for 3h) treatments. In both cell lines, acute EPA treatment did not alter SCD desaturation indices, whereas the acute-on-chronic treatment affected these surrogate markers of SCD activity. This was associated with reduced sterol regulatory-element binding protein-1c and SCD mRNA levels. In contrast, acute and acute-on-chronic DSA treatments significantly reduced SCD desaturation indices without affecting SCD mRNA levels in Caco-2 cells. The present study on intestinal cells shows that the conversion rate of TVA to c9, t11-CLA is affected by other fatty acids present in the diet such as EPA, confirming previous observations in hepatic and mammary cell models.

scholarly journals Forkhead transcription factor FOXO1A is critical for induction of human decidualization

2006 ◽  
Vol 189 (1) ◽  
pp. 179-187 ◽  
Author(s):  
L Grinius ◽  
C Kessler ◽  
J Schroeder ◽  
S Handwerger
Keyword(s):  
Transcription Factor ◽  
Protein Expression ◽  
Critical Role ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Marker Genes ◽  
Fibroblast Cells ◽  
Mrna Levels ◽  
Forkhead Transcription Factor ◽  
Identical Manner

Experiments utilizing RNA interference technology were performed to determine whether the forkhead transcription factor FOXO1A, a member of the FOXO family of proteins, plays a critical role in the induction of human uterine decidualization. Human decidual fibroblast cells were decidualized in vitro for 6 days with medroxyprogester-one, estradiol, and dibutyryl cAMP in the presence or absence of a highly specific FOXO1A small interfering RNA (siRNA) that inhibits FOXO1A mRNA and protein expression by more than 80%. RNA and proteins were extracted from the cells at 0, 2, 4, and 6 days. FOXO1A and IGFBP-1 proteins were determined by immunoblotting; and intracellular mRNA levels for several decidualization marker genes were determined by real-time PCR. Exposure of the cells to FOXO1A siRNA in five separate experiments resulted in a 40–75% inhibition of prolactin, IGFBP-1, tissue inhibitor of metalloproteinase 3 (TIMP3), somatostatin and endometrial bleeding-associated factor (EBAF) mRNAs, all of which are markedly induced during the decidualization process. In contrast, actin and GAPDH mRNA levels did not change during decidualization. The inhibition of mRNA levels was first noted at day 2 and persisted for the remainder of each experiment. Western blot analysis indicated that the FOXO1A siRNA inhibited IGFBP-1 protein expression by 60–80%. Decidual fibroblast cells exposed in an identical manner to a control RNA that had no effect on FOXO1A expression caused only a 0–15% inhibition of the marker genes and IGFBP-1 protein. Taken together, these findings strongly suggest a critical role for FOXO1A in the induction of human decidualization.

scholarly journals Stearoyl-coenzyme A desaturase 1 is required for lipid droplet formation in pig embryo

Reproduction ◽  
2019 ◽  
Vol 157 (3) ◽  
pp. 235-243 ◽  
Author(s):  
Dong-Kyung Lee ◽  
Kwang-Hwan Choi ◽  
Jae Yeon Hwang ◽  
Jong-Nam Oh ◽  
Seung-Hun Kim ◽  
...  
Keyword(s):  
Oleic Acid ◽  
Lipid Bilayers ◽  
Embryo Development ◽  
Coenzyme A ◽  
Mammalian Species ◽  
Formation Rate ◽  
Metabolic Energy ◽  
Mrna Levels ◽  
Parthenogenetic Embryos

Lipid droplets (LD) provide a source of energy, and their importance during embryogenesis has been increasingly recognized. In particular, pig embryos have larger amounts of intercellular lipid bilayers than other mammalian species, suggesting that porcine embryos are more dependent on lipid metabolic pathways. The objective of the present study was to detect the effect of stearoyl-coenzyme A desaturase 1 (SCD1) on LD formation and to associate these effects with the mRNA abundance of LD formation-related genes (SREBP, ARF1, COPG2, PLD1 and ERK2) in in vitro-produced porcine embryos. To determine the effect of SCD1 on LD formation and related genes, we examined the effects of SCD1 inhibition using CAY10566 (an SCD1 inhibitor, 50 μM) on parthenogenetic embryos. SCD1 inhibition downregulated the mRNA levels of LD formation-related genes and embryo development. Our results revealed that SCD1 functions in the regulation of LD formation via phospholipid formation and embryo development. In addition, we treated parthenogenetic embryos with oleic acid (100 μM), which led to a significant increase in the blastocyst formation rate, LD size and number compared to controls. Remarkably, the adverse effects of the SCD1 inhibitor could be counteracted by oleic acid. These data suggest that porcine embryos can use exogenous oleic acid as a metabolic energy source.

TetR-family transcriptional repressor Thermus thermophilus FadR controls fatty acid degradation

Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1589-1601 ◽  
Author(s):  
Yoshihiro Agari ◽  
Kazuko Agari ◽  
Keiko Sakamoto ◽  
Seiki Kuramitsu ◽  
Akeo Shinkai
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Target Genes ◽  
Thermus Thermophilus ◽  
Acyl Chain ◽  
Metabolic Energy ◽  
Straight Chain ◽  
Fatty Acid Degradation ◽  
Acid Degradation

In the extremely thermophilic bacterium Thermus thermophilus HB8, one of the four TetR-family transcriptional regulators, which we named T. thermophilus FadR, negatively regulated the expression of several genes, including those involved in fatty acid degradation, both in vivo and in vitro. T. thermophilus FadR repressed the expression of the target genes by binding pseudopalindromic sequences covering the predicted −10 hexamers of their promoters, and medium-to-long straight-chain (C10–18) fatty acyl-CoA molecules were effective for transcriptional derepression. An X-ray crystal structure analysis revealed that T. thermophilus FadR bound one lauroyl (C12)-CoA molecule per FadR monomer, with its acyl chain moiety in the centre of the FadR molecule, enclosed within a tunnel-like substrate-binding pocket surrounded by hydrophobic residues, and the CoA moiety interacting with basic residues on the protein surface. The growth of T. thermophilus HB8, with palmitic acid as the sole carbon source, increased the expression of FadR-regulated genes. These results indicate that in T. thermophilus HB8, medium-to-long straight-chain fatty acids can be used for metabolic energy under the control of FadR, although the major fatty acids found in this strain are iso- and anteiso-branched-chain (C15 and 17) fatty acids.

scholarly journals Sestrin 2 Attenuates Rat Hepatic Stellate Cell (HSC) Activation and Liver Fibrosis via an mTOR/AMPK-Dependent Mechanism

2018 ◽  
Vol 51 (5) ◽  
pp. 2111-2122 ◽  
Author(s):  
Yi-Bing Hu ◽  
Xiao-Ting Ye ◽  
Qing-Qing Zhou ◽  
Rong-Quan Fu
Keyword(s):  
Liver Fibrosis ◽  
Protein Expression ◽  
Hepatic Fibrosis ◽  
Transforming Growth Factor ◽  
Histopathological Examination ◽  
Stellate Cell ◽  
Smooth Muscle Actin ◽  
Mrna Levels ◽  
Alpha Smooth Muscle Actin

Background/Aims: Sestrin 2 is associated with the pathophysiology of several diseases. The aim of this study was to investigate the effects and potential mechanisms of Sestrin 2 in rat hepatic stellate cells (HSCs) during liver fibrogenesis. Methods: In this study, Sestrin 2 protein expression was detected in rat HSC-T6 cells challenged with transforming growth factor-β (TGF-β) and in mice treated with carbon tetrachloride (CCl4), a well-known model of hepatic fibrosis. Next, HSC-T6 cells and fibrotic mice were transfected with lentivirus. The mRNA expression levels of markers of liver fibrosis [alpha-smooth muscle actin (α-SMA) and collagen 1A1 (Col1A1)] were analyzed by quantitative reverse transcription–polymerase chain reaction (RT-PCR). Cell death and proliferation were evaluated by the MTT assay, and biochemical markers of liver damage in serum [alanine transaminase (ALT) and aspartate transaminase (AST)] were also measured using a biochemical analyzer. Histopathological examination was used to evaluate the degree of liver fibrosis, and protein expression [phospho-adenosine monophosphate-activated protein kinase (p-AMPK), AMPK, phospho-mammalian target of rapamycin (p-mTOR), and mTOR] was determined by western blotting. Results: We found that Sestrin 2 was elevated in both the HSC-T6 cell and hepatic fibrosis models. In vitro, overexpression of Sestrin 2 attenuated the mRNA levels of α-SMA and Col1A1, suppressed α-SMA protein expression, and modulated HSC-T6 cell proliferation. In vivo, overexpression of Sestrin 2 reduced the ALT and AST levels as well as the α-SMA and Col1A1 protein expression in the CCl4 model of liver fibrosis. Moreover, the degree of liver fibrosis was ameliorated. Interestingly, overexpression of Sestrin 2 increased p-AMPK but decreased p-mTOR protein expression. Conclusion: Our findings indicate that Sestrin 2 may attenuate the activation of HSCs and ameliorate liver fibrosis, most likely via upregulation of AMPK phosphorylation and suppression of the mTOR signaling pathway.

scholarly journals Maturation of Human Long-lived Plasma Cells Results in Resistance to Apoptosis by Transcriptional and Epigenetic Regulation

2021 ◽  
Author(s):  
Chester J Joyner ◽  
Ariel Ley ◽  
Doan Nguyen ◽  
Muhammad Ali ◽  
Alessia Corrado ◽  
...  
Keyword(s):  
Gene Expression ◽  
Plasma Cells ◽  
The Novel ◽  
Epigenetic Changes ◽  
Apoptosis Pathway ◽  
Apoptotic Gene ◽  
Survival Gene ◽  
Distinct Morphology ◽  
Antibody Secreting Cells

Antibody secreting cells (ASC) circulate after vaccination and migrate to the bone marrow (BM) where a subset known as long-lived plasma cells (LLPC) persist and secrete antibodies for a lifetime. The mechanisms of how circulating ASC become LLPC are not well elucidated. Here, we show that human blood ASCs have distinct morphology, transcriptomes, and epigenetics compared to BM LLPC. LLPC acquire transcriptional and epigenetic changes in the apoptosis pathway to support their survival. Upregulation of pro-survival gene expression accompanies downregulation of pro-apoptotic gene expression in LLPC. While pro-apoptotic gene loci are less accessible, pro-survival gene loci are not always accompanied by accessibility changes. Importantly, we show similar LLPC morphological and transcriptional maturation of blood ASC in response to the novel in vitro BM mimetic. In all, our study demonstrates that blood ASC in the BM microniche must undergo morphological and molecular changes to mature into apoptotic-resistant LLPC.

Effect of Dietary Vegetable Oil Supplementation on C18 Fatty Acids and Conjugated Linoleic Acid Production; An In vitro Fermentation Study

2013 ◽  
Vol 12 (6) ◽  
pp. 516-520 ◽  
Author(s):  
Julakorn Panatuk ◽  
Suthipong Uriyapongs ◽  
Chainarong Nawanukraw ◽  
Chirasak Phoemchala ◽  
Pitukpol Pornanake
Keyword(s):  
Fatty Acids ◽  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Vegetable Oil ◽  
Acid Production ◽  
In Vitro Fermentation

scholarly journals Aldosterone up-regulates voltage-gated potassium currents and NKCC1 protein membrane fractions

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Parveen Bazard ◽  
Bo Ding ◽  
Harish K. Chittam ◽  
Xiaoxia Zhu ◽  
Thomas A. Parks ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mineralocorticoid Receptor ◽  
Therapeutic Potential ◽  
Potassium Currents ◽  
Chloride Ions ◽  
Mrna Levels ◽  
Mineralocorticoid Receptor Antagonist ◽  
Voltage Gated ◽  
Age Related

Abstract Na+–K+–2Cl− Cotransporter (NKCC1) is a protein that aids in the active transport of sodium, potassium, and chloride ions across cell membranes. It has been shown that long-term systemic treatment with aldosterone (ALD) can enhance NKCC1 protein expression and activity in the aging cochlea resulting in improved hearing. In the present work, we used a cell line with confirmed NKCC1 expression to demonstrate that in vitro application of ALD increased outward voltage-gated potassium currents significantly, and simultaneously upregulated whole lysate and membrane portion NKCC1 protein expression. These ALD-induced changes were blocked by applying the mineralocorticoid receptor antagonist eplerenone. However, application of the NKCC1 inhibitor bumetanide or the potassium channel antagonist Tetraethyl ammonium had no effect. In addition, NKKC1 mRNA levels remained stable, indicating that ALD modulates NKCC1 protein expression via the activation of mineralocorticoid receptors and post-transcriptional modifications. Further, in vitro electrophysiology experiments, with ALD in the presence of NKCC1, K+ channel and mineralocorticoid receptor inhibitors, revealed interactions between NKCC1 and outward K+ channels, mediated by a mineralocorticoid receptor-ALD complex. These results provide evidence of the therapeutic potential of ALD for the prevention/treatment of inner ear disorders such as age-related hearing loss.

34 EFFECTS OF L-CARNITINE AND trans-10,cis-12 CONJUGATED LINOLEIC ACID SUPPLEMENTATION DURING MATURATION ON DEVELOPMENT AND CRYOTOLERANCE OF BOVINE EMBRYOS PRODUCED IN VITRO

2016 ◽  
Vol 28 (2) ◽  
pp. 147
Author(s):  
J. Block ◽  
A. M. Zolini ◽  
E. Carrascal-Triana ◽  
A. Ruiz ◽  
P. J. Hansen ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Embryo Development ◽  
Blastocyst Stage ◽  
Cleavage Rate ◽  
Sodium Pyruvate ◽  
Maturation Medium ◽  
Fetal Bovine ◽  
Hatching Rates

The objective of the present study was to determine the effect of supplementation of maturation media with L-carnitine and trans-10,cis-12 conjugated linoleic acid (CLA) on embryo development and survival following cryopreservation. Immature bovine cumulus-oocyte complexes (n = 1796) were harvested from abattoir-derived ovaries and randomly assigned in a 2 × 2 factorial design to be matured in maturation medium [TCM-199 with Earle salts supplemented with 10% (vol/vol) bovine steer serum, 2 μg mL–1 oestradiol 17-β, 20 μg mL–1 bovine FSH, 22 μg mL–1 sodium pyruvate, 50 μg mL–1 gentamicin sulfate, and 1 mM glutamax®] supplemented with or without 100 mM CLA and with or without 3.03 mM L-carnitine for 22 to 24 h at 38.5°C in a humidified atmosphere of 5% CO2. The proportion of oocytes that cleaved was determined on Day 3 after insemination, and the proportion of oocytes developing to the blastocyst and advanced blastocysts stages (expanded, hatching, and hatched) was assessed on Day 7. Blastocyst and expanded blastocyst stage embryos (n = 270) were harvested on Day 7 and subjected to controlled-rate freezing following equilibration in 1.5 M ethylene glycol. Embryos were thawed and then cultured for 72 h in SOF-BE1 (Fields et al. 2011) supplemented with 10% (vol/vol) fetal bovine serum and 50 μM dithiothreitol. Post-thaw re-expansion and hatching rates were determined at 24, 48, and 72 h. The experiment was replicated 5 times. There was no effect of supplementation of maturation medium with either CLA or L-carnitine on the proportion of oocytes that cleaved at Day 3 or that developed to the blastocyst and advanced blastocyst stages at Day 7 after insemination. There was no interaction between CLA and L-carnitine affecting cleavage rate or embryo development. Supplementation of maturation medium with L-carnitine did not affect post-thaw re-expansion or hatching rates. In contrast, treatment with CLA during maturation reduced (P < 0.05) post-thaw re-expansion (24 h: 75.2 ± 3.8% v. 60.3 ± 4.1%; 48 h: 82.0 ± 3.4% v. 64.9 ± 4.0%; 72 h: 78.9 ± 3.6% v. 65.9 ± 4.0%, respectively) and hatching (24 h: 33.7 ± 4.2% v. 23.5 ± 3.6%; 48 h: 61.1 ± 4.3% v. 44.0 ± 4.2%; 72 h: 62.6 ± 4.3% v. 50.2 ± 4.2%, respectively) rates at all time points. There was no interaction between CLA and L-carnitine affecting post-thaw viability. In conclusion, supplementation of maturation medium with L-carnitine did not affect embryo development or post-thaw viability. Although addition of CLA during maturation did not affect embryo development, post-thaw cryotolerance was reduced following CLA supplementation. There was no beneficial effect of supplementing maturation medium with both CLA and L-carnitine on embryo development or post-thaw cryosurvival.

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