Palmitic acid methyl ester induces cardiac hypertrophy through activating the GPR receptor-mediated changes of intracellular calcium concentrations and mitochondrial functions.

Myocardial hypertrophy is associated with a significant increase in intracellular Ca2+ , which can be induced by long-chain fatty acid. Palmitic acid methyl ester (PAME), a fatty acid ester released from adipose tissue, superior cervical ganglion, and retina, has been found to have anti-inflammation, antifibrosis, and peripheral vasodilation effects. However, the effects of PAME on cardiomyocytes are still unclear. The aim of this study was to determine whether PAME could disrupt the intracellular Ca2+ balance, leading to cardiomyocyte hypertrophy. Neonatal rat cardiomyocytes were treated with various concentrations (10-100 µM) of PAME for 1-4 days. Cytosolic Ca2+ and mitochondrial Ca2+ concentrations were examined using Fura-2 AM and Rhod-2, respectively. After treatment with PAME for 4 days, mitochondrial Ca2+ , an indicator of the state of mitochondrial permeability transition pore (MPTP), and cell death were monitored by flow cytometric analysis. ATP levels were detected using the ATP assay kit. Cardiomyocyte hypertrophy was analyzed by measuring the cardiac hypertrophy biomarker and cell area using quantitative real time-polymerase chain reaction, Western Blot analysis and immunofluorescence analysis. Our results show that PAME concentration- and time-dependently increased cytosolic and mitochondria Ca2+ through the mitochondrial calcium uniporter. Moreover, treatment with PAME for 4 days caused MPTP opening, thereby reducing ATP production and enhancing reactive oxygen species (ROS) generation, and finally led to cardiomyocyte hypertrophy. These effects caused by PAME treatment were attenuated by the G-protein coupled receptor 40 (GPR40) inhibitor. In conclusion, PAME impaired mitochondrial function, which in turn led to cardiomyocyte hypertrophy through increasing the mitochondrial Ca2+ levels mediated by activating the GPR40 signaling pathway.

Intermittent Hypoxia Inhibits Na+-H+ Exchange-Mediated Acid Extrusion Via Intracellular Na+ Accumulation in Cardiomyocytes.

BACKGROUND/AIMS: Intermittent hypoxia (IH) has been shown to exert preconditioning-like cardioprotective effects. It also has been reported that IH preserves intracellular pH (pHi) during ischemia and protects cardiomyocytes against ischemic reperfusion injury. However, the exact mechanism is still unclear. METHODS: In this study, we used proton indicator BCECF-AM to analyze the rate of pHi recovery from acidosis in the IH model of rat neonatal cardiomyocytes. Neonatal cardiomyocytes were first treated with repetitive hypoxia-normoxia cycles for 1-4 days. Cells were then acid loaded with NH4Cl, and the rate of pHi recovery from acidosis was measured. RESULTS: We found that the pHi recovery rate from acidosis was much slower in the IH group than in the room air (RA) group. When we treated cardiomyocytes with Na+-H+ exchange (NHE) inhibitors (Amiloride and HOE642) or Na+-free Tyrode solution during the recovery, there was no difference between RA and IH groups. We also found intracellular Na+ concentration ([Na+]i) significantly increased after IH exposure for 4 days. However, the phenomenon could be abolished by pretreatment with ROS inhibitors (SOD and phenanathroline), intracellular calcium chelator or Na+-Ca2+ exchange (NCX) inhibitor. Furthermore, the pHi recovery rate from acidosis became faster in the IH group than in the RA group when inhibition of NCX activity. CONCLUSIONS: These results suggest that IH would induce the elevation of ROS production. ROS then activates Ca2+-efflux mode of NCX and results in intracellular Na+ accumulation. The rise of [Na+]i further inhibits the activity of NHE-mediated acid extrusion and retards the rate of pHi recovery from acidosis during IH.

Nestedness Maximization in Complex Networks through the Fitness-Complexity Algorithm.

Nestedness refers to the structural property of complex networks that the neighborhood of a given node is a subset of the neighborhoods of better-connected nodes. Following the seminal work by Patterson and Atmar (1986), ecologists have been long interested in revealing the configuration of maximal nestedness of spatial and interaction matrices of ecological communities. In ecology, the BINMATNEST genetic algorithm can be considered as the state-of-the-art approach for this task. On the other hand, the fitness-complexity ranking algorithm has been recently introduced in the economic complexity literature with the original goal to rank countries and products in World Trade export networks. Here, by bringing together quantitative methods from ecology and economic complexity, we show that the fitness-complexity algorithm is highly effective in the nestedness maximization task. More specifically, it generates matrices that are more nested than the optimal ones by BINMATNEST for 61.27% of the analyzed mutualistic networks. Our findings on ecological and World Trade data suggest that beyond its applications in economic complexity, the fitness-complexity algorithm has the potential to become a standard tool in nestedness analysis.

Non-lethal levels of oxidative stress in response to short-term intermittent hypoxia enhance ca²âº handling in neonatal rat cardiomyocytes.

BACKGROUND/AIMS: Intermittent hypoxia (IH) may exert pre-conditioning-like cardioprotective effects and alter Ca(2+) regulation; however, the exact mechanism of these effects remains unclear. Thus, we examined Ca(2+)-handling mechanisms induced by IH in rat neonatal cardiomyocytes. METHODS: Cardiomyocytes were exposed to repetitive hypoxia-re-oxygenation cycles for 1-4 days. Mitochondrial reactive oxygen species (ROS) generation was determined by flow cytometry, and intracellular Ca(2+) concentrations were measured using a live-cell fluorescence imaging system. Protein kinase C (PKC) isoforms and Ca(2+)-handling proteins were analysed using immunofluorescence and western blotting. RESULTS: After IH exposure for 4 days, the rate of Ca(2+) extrusion from the cytosol to the extracellular milieu during 40-mM KCl-induced Ca(2+) mobilization increased significantly, whereas ROS levels increased mildly. IH activated PKC isoforms, which translocated to the membrane from the cytosol, and Na(+)/Ca(2+) exchanger-1, leading to enhanced Ca(2+) efflux capacity. Simultaneously, IH increased sarcoplasmic reticulum (SR) Ca(2+)-ATPase and ryanodine receptor 2 (RyR-2) activities and RyR-2 expression, resulting in improved Ca(2+) uptake and release capacity of SR in cardiomyocytes. CONCLUSIONS: IH-induced mild elevations in ROS generation can enhance Ca(2+) efflux from the cytosol to the extracellular milieu and Ca(2+)-mediated SR regulation in cardiomyocytes, resulting in enhanced Ca(2+)-handling ability.

Intermittent hypoxia-induced protein phosphatase 2A activation reduces PC12 cell proliferation and differentiation.

BACKGROUND: Intermittent hypoxia (IH) plays a critical role in sleep breathing disorder-associated hippocampus impairments, including neurocognitive deficits, irreversible memory and learning impairments. IH-induced neuronal injury in the hippocampus may result from reduced precursor cell proliferation and the relative numbers of postmitotic differentiated neurons. However, the mechanisms underlying IH-induced reactive oxygen species (ROS) generation effects on cell proliferation and neuronal differentiation remain largely unknown. RESULTS: ROS generation significantly increased after 1-4 days of IH without increased pheochromocytoma-12 (PC12) cell death, which resulted in increased protein phosphatase 2A (PP2A) mRNA and protein levels. After 3-4 days of IH, extracellular signal-regulated kinases 1/2 (ERK1/2) protein phosphorylation decreased, which could be reversed by superoxide dismutase (SOD), 1,10-phenanthroline (Phe), the PP2A phosphorylation inhibitors, okadaic acid (OKA) and cantharidin, and the ERK phosphorylation activator nicotine (p < 0.05). In particular, the significantly reduced cell proliferation and increased proportions of cells in the G0/G1 phase after 1-4 days of IH (p < 0.05), which resulted in decreased numbers of PC12 cells, could be reversed by treatment with SOD, Phe, PP2A inhibitors and an ERK activator. In addition, the numbers of nerve growth factor (NGF)-induced PC12 cells with neurite outgrowths after 3-4 days of IH were less than those after 4 days of RA, which was also reversed by SOD, Phe, PP2A inhibitors and an ERK activator. CONCLUSIONS: Our results suggest that IH-induced ROS generation increases PP2A activation and subsequently downregulates ERK1/2 activation, which results in inhibition of PC12 cell proliferation through G0/G1 phase arrest and NGF-induced neuronal differentiation.

Serum Malondialdehyde-Oxidized Low-Density Lipoprotein Level Is Associated with Arterial Stiffness by Cardio-Ankle Vascular Index in Coronary Artery Bypass Graft Patients.

A high malondialdehyde-oxidized low-density lipoprotein (MDA-oxLDL) level is associated with atherosclerotic cardiovascular diseases and major adverse cardiovascular events. A higher cardio-ankle vascular index (CAVI) is independently associated with an increased risk of cardiovascular events, cardiovascular mortality, myocardial infarction, and stroke in patients with cardiovascular risk. Thus, this study aimed to evaluate the relationship between serum MDA-oxLDL levels and CAVI in patients with triple-vessel coronary artery disease who underwent coronary artery bypass graft (CABG) surgery. Fasting blood samples and baseline characteristics were obtained from 88 patients who had undergone CABG. A commercialized enzyme-linked immunosorbent assay was used to measure MDA-oxLDL levels. An automatic pulse wave analyzer was used to measure CAVI values, and each side of CAVI values of ≥9 was designated as arterial stiffness. In total, 47 participants were assigned to the arterial stiffness group. More patients had diabetes mellitus, were older, and had higher serum MDA-oxLDL levels in the arterial stiffness group than in the control group. A multivariate logistic regression analysis disclosed that MDA-oxLDL and diabetes mellitus were independent predictors of arterial stiffness. Moreover, according to the Spearman's correlation analysis, the serum MDA-oxLDL level was positively associated with both left and right CAVI. Serum MDA-oxLDL levels were positively associated with arterial stiffness in patients who had undergone CABG.

Baicalein and luteolin inhibit ischemia/reperfusion-induced ferroptosis in rat cardiomyocytes.

BACKGROUND: Ischemia/reperfusion (I/R) is associated with severe cellular damage and death. Ferroptosis, a new form of regulated cell death caused by the accumulation of iron-mediated lipid peroxidation, has been found in several diseases including I/R injury, which was reported to be suppressed by flavonoids. Baicalein (BAI) and luteolin (Lut) are flavonoids and were shown to reduce the myocardial I/R injury. BAI was found to suppress ferroptosis in cancer cells via reducing reactive oxygen species (ROS) generation. However, the anti-ferroptosis effect of Lut on ferroptosis has not been reported. This study aimed to investigate whether ferroptosis reduction contributes to the BAI- and Lut-protected cardiomyocytes. METHODS: This research used erastin, RSL3, and Fe-SP to induce ferroptosis. Cell viability was examined using MTT assay. Annexin V-FITC, CM-H2DCFDA, and Phen Green SK diacetate (PGSK) fluorescent intensity were detected to analyze apoptotsis, ROS levels, and Fe2+ concentrations, respectively. qPCR and Western blot analysis were conducted to detect the levels of mRNA and protein, respectively. RESULTS: Our data show that BAI and Lut protected cardiomyocytes against ferroptosis caused by ferroptosis inducers and I/R. Moreover, both BAI and Lut decreased ROS and malondialdehyde (MDA) generation and the protein levels of ferroptosis markers, and restored Glutathione peroxidase 4 (GPX4) protein levels in cardiomyocytes reduced by ferroptosis inducers. BAI and Lut reduced the I/R-induced myocardium infarction and decreased the levels of Acsl4 and Ptgs2 mRNA. CONCLUSIONS: BAI and Lut could protect the cardiomyocytes against the I/R-induced ferroptosis via suppressing accumulation of ROS and MDA.

Licochalcone a improves cardiac functions after ischemia-reperfusion via reduction of ferroptosis in rats.

Myocardial ischemia-reperfusion (I/R) injury triggers several cell death types, including apoptosis, autophagy, and ferroptosis. Licochalcone A (LCA), a natural flavonoid compound isolated from the root of Glycyrrhiza glabra, has been demonstrated to exert potential pharmacological benefits, such as antioxidant, antitumor, and anti-inflammatory activities. The present study aimed to investigate the involvement of ferroptosis in the pathogenesis of I/R and determine whether LCA can inhibit ferroptosis to prevent the myocardial I/R injury in rats. The effects of LCA on myocardial I/R injury were detected by examining the left ventricular-developed pressure and triphenyltetrazolium chloride staining. We conducted Western blotting analyses, ELISA assay, and quantitative real-time PCR to determine the levels of ferroptosis-related molecules. To demonstrate the cardioprotective effect of LCA in vitro, H9c2 and primary neonatal rat cardiomyocytes were co-treated with ferroptosis inducers (erastin, RSL3, or Fe-SP) and LCA for 16 and 24 h. Our ex vivo study showed that LCA increased the cardiac contractility, and reduced the infarct volume and ferroptosis-related biomarkers in rat hearts after I/R. Moreover, LCA reduced the levels of ferroptosis inducers-induced reactive oxygen species generation, lipid peroxidation, and ferroptosis-related biomarkers in cultured H9c2 cells and cardiomyocytes. LCA also reduced the Fe-SP-increased nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 protein levels in cultured cardiomyocytes. In the present study, we showed that the LCA-induced cardioprotective effects in attenuating the myocardial I/R injury were correlated with ferroptosis regulation, and provided a possible new therapeutic strategy for prevention or therapy of the myocardial I/R injury.

PVP/CS/Phyllanthus emblica Nanofiber Membranes for Dry Facial Masks: Manufacturing Process and Evaluations.

In the wake of increasing demands on skin health, we propose simple, natural, and safe dry facial masks that restrict melanin synthesis. Phyllanthus emblica (P. emblica) is made into powders via a low-temperature extraction and freeze-drying process to serve as a natural agent. Next, it is added to mixtures containing Polyvinylpyrrolidone (PVP) and Chitosan (CS), after which the blends are electrospun into PVP/CS/P. emblica nanofiber membrane dry facial masks using the electrospinning technique. The dry facial masks are evaluated using the calibration analysis method, extraction rate test, scanning electron microscopy (SEM), release rate test, tyrosinase inhibition assay, biocompatibility test, and anti-inflammatory capacity test. Test results indicate that when the electrospinning mixture contains 29.0% P. emblica, the nanofibers have a diameter of ≤214.27 ± 74.51 nm and a water contact angle of 77.25 ± 2.21. P. emblica is completely released in twenty minutes, and the tyrosinase inhibition rate reaches 99.53 ± 0.45% and the cell activity ≥82.60 ± 1.30%. Moreover, the anti-inflammatory capacity test results suggest that dry facial masks confine inflammatory factors. PVP/CS/P. emblica nanofiber dry facial masks demonstrate excellent tyrosinase inhibition and are hydrophilic, biocompatible, and inflammation-free. The dry facial masks are a suitable material that is worthwhile exploring and applying to the cosmetic field.

Age and Serum Adipocyte Fatty-Acid-Binding Protein Level Are Associated with Aortic Stiffness in Coronary Artery Bypass Graft Patients.

Old age has been proven to be related to progressed arterial or aortic stiffness. Aortic stiffness is an independent predictor of all-cause and cardiovascular disease mortalities in patients who have undergone coronary artery bypass grafting (CABG) surgery. Higher serum concentrations of adipocyte fatty-acid-binding protein (A-FABP) could be considered a predictor of aortic stiffness in patients with hypertension or diabetes mellitus. This study aims to investigate the relationships between A-FABP and aortic stiffness in patients who have received CABG. A total of 84 CABG patients were enrolled in our study from September 2018 to May 2019. Serum A-FABP levels were determined using a commercial enzyme immunoassay. Carotid−femoral pulse wave velocity (cfPWV) > 10 m/s was defined as aortic stiffness. Of the 84 CABG patients, 28 (33.3%) with aortic stiffness had a higher average age; exhibited higher rates of diabetes; and had higher serum creatinine, C-reactive protein, and A-FABP levels compared to controls. Multivariable logistic regression revealed that serum A-FABP levels (odds ratio (OR) = 1.068, 95% confidence interval (CI) 1.017−1.121, p = 0.008) and age (OR = 1.204, 95% CI 1.067−1.359, p = 0.003) were independent predictors of aortic stiffness. Multivariable stepwise linear regression revealed significant positive correlations of age and A-FABP levels with cfPWV values. Serum A-FABP level is positively correlated with cfPWV values, and a high serum A-FABP level is associated with aortic stiffness in patients who have undergone CABG.

Berberine protects cardiac cells against ferroptosis.

Objectives: Cardiovascular diseases are one of the primary causes of death. Cardiomyocyte loss is a significant feature of cardiac injury. Ferroptosis is iron-dependent cell death, which occurs due to excess iron and reactive oxygen species (ROS) accumulation causing lipid peroxidation, and subsequent cell death. Ferroptosis has been confirmed to mediate ischemia/reperfusion-induced cardiomyopathy and chemotherapy-induced cardiotoxicity. Berberine (BBR) has been proven to protect the heart from cardiomyopathies, including cardiac hypertrophy, heart failure, myocardial infarction, and arrhythmias. It protects cardiomyocytes from apoptosis and autophagy. However, the relation between BBR and ferroptosis is still unknown. This study aimed to confirm if BBR reduces cardiac cell loss via inhibiting ferroptosis. Materials and Methods: We used erastin and Ras-selective lethal small molecule 3 (RSL3) to establish a ferroptosis model in an H9c2 cardiomyoblast cell line and rat neonatal cardiomyocytes to prove that BBR has a protective effect on cardiac cells via inhibiting ferroptosis. Results: In H9c2 cardiomyoblasts, the results showed that BBR reduced erastin and RSL3-induced cell viability loss. Moreover, BBR decreased ROS accumulation and lipid peroxidation in cells induced with ferroptosis. Furthermore, quantitative polymerase chain reaction results showed that Ptgs2 mRNA was reduced in BBR-treated cells. In rat neonatal cardiomyocytes, BBR reduced RSL3-induced loss of cell viability. Conclusion: These results indicated that BBR inhibited ferroptosis via reducing ROS generation and reducing lipid peroxidation in erastin and RSL3-treated cardiac cells.

A Study on Preparation and Property Evaluations of Composites Consisting of TPU/Triclosan Membranes and Tencel®/LMPET Nonwoven Fabrics.

This study investigated eco-friendly antibacterial medical protective clothing via the nonwoven process and characteristic evaluations. Firstly, Tencel® fibers and low melting point polyester (LMPET) fibers (re-sliced and granulated from recycled PET bottles) were mixed at different ratios and then needle punched at diverse needle rolling depths. The influences of manufacturing parameters on the Tencel®/LMPET nonwoven fabrics were examined in terms of mechanical properties, water vapor transmission rate, and stiffness. Next, Tencel®/LMPET nonwoven fabrics were combined with thermoplastic polyurethane (TPU)/Triclosan antibacterial membranes that contained different contents of triclosan using melt processing technology. The resulting Tencel®/LMPET/TPU/Triclosan composites were characterized via different measurements; an optimal bursting strength of 86.86 N, an optimal horizontal tensile strength of 41.90 N, and an optimal stiffness along the MD and CD of 8.60 cm were recorded. Furthermore, the Tencel®/LMPET/TPU/Triclosan composites exhibited a distinct inhibition zone in the antibacterial measurement, and the hydrostatic pressure met the requirements of the EN 14126:2003 and GB 19082-200 disposable medical protective gear test standards.

A Study on Highly Effective Electromagnetic Wave Shield Textile Shell Fabrics Made of Point Polyester/Metallic Core-Spun Yarns.

In this study, stainless steel (SS) filaments are wrapped in Ge fibers to form core-spun yarns. The yarns along with 500 D polyester (PET) fibers undergo weaving, thereby forming functional woven fabrics. The experiment is composed of two parts:yarns and fabrics. The yarns are twisted with TPI of 8, 9, 10, 11, and 12, and then tested for tensile strength and tensile elongation. The yarns possess mechanical properties that are dependent on the TPI-the higher the TPI, the better the mechanical properties. The maximal mechanical properties occur when the core-spun yarns are made of 12 TPI where the maximal tensile strength is 5.26 N and the lowest elongation is 43.2%. As for the functional woven fabrics, they are made of Ge/SS core-spun yarns as the weft yarns and 500 D PET yarns as the warp yarns. The tensile strength, tensile elongation, negative ion release, electromagnetic interference shielding effectiveness (EMI SE), and air permeability tests are conducted, determining the optimal woven fabrics. The 12 TPI core-spun yarns provide the woven fabrics with the maximal tensile strength of 153.6 N and the optimal elongation at break of 10.08%. In addition, the woven fabrics made with 8 or 9 TPI core-spun yarns exhibit an optimal EMI SE of 41 dB, an optimal air permeability of 212 cm3/cm2/s, and an optimal release amount of negative ion of 550-600 ions/cc. The proposed woven fabrics have a broad range of applications, such as functional garments and bedding.

Lay-Up Compound Matrices for Application of Medical Protective Clothing: Manufacturing Techniques and Property Evaluations.

Medical protective clothing is the first line of defense for medical staff, which makes the acquisition of protection and multiple function challenging. When it comes to contagious diseases, the physical properties of protective clothing are deemed the top priority and, subsequently, they have significant meaning for the structural design, production cost evaluation, convenient production, and innovation. In this study, nonwoven technology is employed to produce matrices in which mechanical properties are supported by Tencel fibers and recycled Kevlar fibers. Next, the electrostatic spinning is conducted to generate breathable and waterproof films. The nonwoven fabrics and membranes are combined to have diverse functions, forming lay-up compound matrices for medical protective clothing. Moreover, measurements are conducted to characterize the lay-up compound matrices in terms of the tensile strength, tearing strength, bursting strength, puncture resistance, stiffness, air-permeable property, surface resistance, comfort performance, sub-micron particulate filtration efficiency, and the penetration of synthetic blood. As for the nonwoven fabrics, the mechanical properties are significantly improved after Kevlar fibers are incorporated. The tensile strength is (62.6 ± 2.4) N along the machine direction (MD) and (50.1 ± 3.1) N along the cross machine direction (CD); the tearing strength is (29.5 ± 1.6) N along the MD and (43.0 ± 1.7) N along the CD; the bursting strength is (365.8 ± 5.0) kPa; and the puncture resistance is (22.6 ± 1.0) N. Moreover, the lay-up compound matrices exhibit a stiffness of (14.7 ± 0.2) cm along the MD and (14.6 ± 0.1) cm along the CD, a surface resistance of (2.85 × 109 ± 0.37 × 109) Ω, an air-permeable property of (45.4 ± 2.3) cm3/s/cm2, and sub-micron particulate filtration efficiency of over 98%. In the measurement for penetration of synthetic blood, the K40/PAN/TPU group prevents the synthetic blood from penetration. Hence, the incorporation of recycled Kevlar fibers and lay-up compound technique creates good physical properties, an appropriate comfort attribute, and functions, which suggests that this study provides a greater diversity and new concepts for the production of medical protective clothing.

Xanthohumol Protects the Rat Myocardium against Ischemia/Reperfusion Injury-Induced Ferroptosis.

Ferroptosis is an iron-dependent form of cell death caused by the inactivation of glutathione peroxidase 4 (GPX4) and accumulation of lipid peroxides. Ferroptosis has been found to participate in the ischemia-reperfusion (I/R) injury, leading to heart dysfunction and myocardial cell death. Xanthohumol (XN), a prenylated flavonoid isolated from Humulus lupulus, has multiple pharmacological activities, such as anti-inflammatory and antioxidant. This study is aimed at investigating whether XN could attenuate the I/R-induced ferroptosis in cardiomyocytes and the underlying mechanisms. Cardiomyocytes were treated with Fe-SP and RSL3, and the rat hearts were treated with I/R. The results from the present study show that XN was able to protect cardiomyocytes against Fe-SP- and RSL3-induced ferroptotic cell death by decreasing the production of lipid peroxidation and ROS, chelating iron, reducing the NRF2 protein level, and modulating the protein levels of GPX4. Moreover, XN significantly decreased the mRNA levels of ferroptosis markers, Ptgs2 and Acsl4, and the protein levels of ACSL4 and NRF2 and modulated the protein levels of GPX4 in I/R-treated hearts. The findings from the present study suggest that XN might have the therapeutic potential for the I/R-induced ferroptosis injury.

Gossypol Acetic Acid Attenuates Cardiac Ischemia/Reperfusion Injury in Rats via an Antiferroptotic Mechanism.

Myocardial ischemia/reperfusion (I/R) injury has been associated with ferroptosis, which is characterized by an iron-dependent accumulation of lipid peroxide to lethal levels. Gossypol acetic acid (GAA), a natural product taken from the seeds of cotton plants, prevents oxidative stress. However, the effects of GAA on myocardial I/R-induced ferroptosis remain unclear. This study investigated the ability of GAA to attenuate I/R-induced ferroptosis in cardiomyocytes along with the underlying mechanisms in a well-established rat model of myocardial I/R and isolated neonatal rat cardiomyocytes. H9c2 cells and cardiomyocytes were treated with the ferroptosis inducers erastin, RSL3, and Fe-SP. GAA could protect H9c2 cells against ferroptotic cell death caused by these ferroptosis inducers by decreasing the production of malondialdehyde and reactive oxygen species, chelating iron content, and downregulating mRNA levels of Ptgs2. GAA could prevent oxygen-glucose deprivation/reperfusion-induced cell death and lipid peroxidation in the cardiomyocytes. Moreover, GAA significantly attenuated myocardial infarct size, reduced lipid peroxidation, decreased the mRNA levels of the ferroptosis markers Ptgs2 and Acsl4, decreased the protein levels of ACSL4 and NRF2, and increased the protein levels of GPX4 in I/R-induced ex vivo rat hearts. Thus, GAA may play a cytoprotectant role in ferroptosis-induced cardiomyocyte death and myocardial I/R-induced ferroptotic cell death.

Palmitic Acid Methyl Ester Enhances Adipogenic Differentiation in Rat Adipose Tissue-Derived Mesenchymal Stem Cells through a G Protein-Coupled Receptor-Mediated Pathway.

Adipogenic differentiation from stem cells has become a research target due to the increasing interest in obesity. It has been indicated that adipocytes can secrete palmitic acid methyl ester (PAME), which is able to regulate stem cell proliferation. However, the effects of PAME on adipogenic differentiation in stem cell remain unclear. Here, we present that the adipogenic differentiation medium supplemented with PAME induced the differentiation of rat adipose tissue-derived mesenchymal stem cells (rAD-MSCs) into adipocyte. rAD-MSCs were treated with PAME for 12 days and then subjected to various analyses. The results from the present study show that PAME significantly increased the levels of adipogenic differentiation markers, PPARγ and Gpd1, and enhanced adipogenic differentiation in rAD-MSCs. Furthermore, the level of GPR40/120 protein increased during induction of adipocyte differentiation in rAD-MSCs. Cotreatment with PAME and a GPR40/120 antagonist together inhibited the PAME-enhanced adipogenic differentiation. Moreover, PAME significantly increased phosphorylation of extracellular signal-regulated kinases (ERK), but not AKT and mTOR. Cotreatment with PAME and a GPR40/120 antagonist together inhibited the PAME-enhanced ERK phosphorylation and adipogenic differentiation. PAME also increased the intracellular Ca2+ levels. Cotreatment with PAME and a Ca2+ chelator or a phospholipase C (PLC) inhibitor prevented the PAME-enhanced ERK phosphorylation and adipogenic differentiation. Our data suggest that PAME activated the GPR40/120/PLC-mediated pathway, which in turn increased the intracellular Ca2+ levels, thereby activating the ERK, and eventually enhanced adipogenic differentiation in rAD-MSCs. The findings from the present study might help get insight into the physiological roles and molecular mechanism of PAME in regulating stem cell differentiation.

Palmitic Acid Methyl Ester Induces G2/M Arrest in Human Bone Marrow-Derived Mesenchymal Stem Cells via the p53/p21 Pathway.

Bone marrow-derived mesenchymal cells (BM-MSCs) are able to differentiate into adipocytes, which can secrete adipokines to affect BM-MSC proliferation and differentiation. Recent evidences indicated that adipocytes can secrete fatty acid metabolites, such as palmitic acid methyl ester (PAME), which is able to cause vasorelaxation and exerts anti-inflammatory effects. However, effects of PAME on BM-MSC proliferation remain unclear. The aim of this study was to investigate the effect of PAME on human BM-MSC (hBM-MSC) proliferation and its underlying molecular mechanisms. hBM-MSCs were treated with PAME for 48 h and then subjected to various analyses. The results from the present study show that PAME significantly reduced the levels of G2/M phase regulatory proteins, cyclin-dependent kinase 1 (Cdk1), and cyclin B1 and inhibited proliferation in hBM-MSCs. Moreover, the level of Mdm2 protein decreased, while the levels of p21 and p53 protein increased in the PAME-treated hBM-MSCs. However, PAME treatment did not significantly affect apoptosis/necrosis, ROS generation, and the level of Cdc25C protein. PAME also induced intracellular acidosis and increased intracellular Ca2+ levels. Cotreatment with PAME and Na+/H+ exchanger inhibitors together further reduced the intracellular pH but did not affect the PAME-induced decreases of cell proliferation and increases of the cell population at the G2/M phase. Cotreatment with PAME and a calcium chelator together inhibited the PAME-increased intracellular Ca2+ levels but did not affect the PAME-induced cell proliferation inhibition and G2/M cell cycle arrest. Moreover, the half-life of p53 protein was prolonged in the PAME-treated hBM-MSCs. Taken together, these results suggest that PAME induced p53 stabilization, which in turn increased the levels of p53/p21 proteins and decreased the levels of Cdk1/cyclin B1 proteins, thereby preventing the activation of Cdk1, and eventually caused cell cycle arrest at the G2/M phase. The findings from the present study might help get insight into the physiological roles of PAME in regulating hBM-MSC proliferation.

Intermittent Hypoxia Induces Autophagy to Protect Cardiomyocytes From Endoplasmic Reticulum Stress and Apoptosis.

Intermittent hypoxia (IH), characterized as cyclic episodes of short-period hypoxia followed by normoxia, occurs in many physiological and pathophysiological conditions such as pregnancy, athlete, obstructive sleep apnea, and asthma. Hypoxia can induce autophagy, which is activated in response to protein aggregates, in the proteotoxic forms of cardiac diseases. Previous studies suggested that autophagy can protect cells by avoiding accumulation of misfolded proteins, which can be generated in response to ischemia/reperfusion (I/R) injury. The objective of the present study was to determine whether IH-induced autophagy can attenuate endoplasmic reticulum (ER) stress and cell death. In this study, H9c2 cell line, rat primary cultured cardiomyocytes, and C57BL/6 male mice underwent IH with an oscillating O2 concentration between 4 and 20% every 30 min for 1-4 days in an incubator. The levels of LC3, an autophagy indicator protein and CHOP and GRP78 (ER stress-related proteins) were measured by Western blotting analyses. Our data demonstrated that the autophagy-related proteins were upregulated in days 1-3, while the ER stress-related proteins were downregulated on the second day after IH. Treatment with H2O2 (100 µM) for 24 h caused ER stress and increased the level of ER stress-related proteins, and these effects were abolished by pre-treatment with IH condition. In response to the autophagy inhibitor, the level of ER stress-related proteins was upregulated again. Taken together, our data suggested that IH could increase myocardial autophagy as an adaptive response to prevent the ER stress and apoptosis.

Preparation of a Periodic Polystyrene Nanosphere Array Using the Dip-Drop Method with Post-deposition Etching and Its Application of Improving Light Extraction Efficiency of InGaN/GaN LEDs.

In this study, we synthesized a periodic polystyrene nanosphere (PS NS) array using the dip-drop method with post-deposition etching to improve the light extraction efficiency (LEE) of InGaN/GaN light-emitting diodes (LEDs). The dip-drop method has advantages such as simple procedure, inexpensive equipment, room temperature deposition, and easy implementation in LEDs. The arrangement of PS NSs on an indium-tin-oxide (ITO)-coated glass substrate depends on the average dip-drop speed and the concentration of the PS NS suspension. The periodic PS NS array can modulate the in-plane wave vector of emission light from a semiconductor to free space and thus increase the escape probability. The calculated and experimental results indicated that the light output intensity of the InGaN/GaN LEDs can be improved by using the periodic PS NS array as a window layer; this array comprises PS NSs with a diameter of 100 nm separated with periods of 100 and 100 nm in the x and y directions. Because of the improved LEE, the InGaN/GaN LEDs with the optimal PS NS array window layers exhibited a 38% increase in light output intensity compared with the conventional InGaN/GaN LEDs under 20-mA driving current.