Oleic Acid Group Research Articles

Camellia euphlebia flower extract inhibits oleic acid-induced lipid accumulation via reduction of lipogenesis in HepG2 cells

IntroductionCamellia euphlebia is a medicinal and edible plant used to treat hepatic disease in Southern China. However, there have been no reports on the anti-obesity and hypolipidemic effects of Camellia euphlebia flower. This study evaluated the hepatic lipid-lowering potential of different preparations of Camellia euphlebia flower extracts using in vitro models. MethodsA model of hepatic steatosis in the HepG2 cells was induced by oleic acid. HepG2 cells were divided into control group, oleic acid group, oleic acid plus aqueous extract group and oleic acid plus ethanol extract group. The cytotoxicity was quantitatively assessed by methyl-thiazolyl-tetrazolium and lactate dehydrogenase release assay. Lipid accumulation and intracellular triglyceride level were also evaluated by oil red O staining and a commercially available assay kit respectively. Expressions of 3-hydroxy-3-methylglutaryl CoA reductase, fatty acid synthase, acetyl-CoA carboxylase and glycerol-3-phosphate acyl transferase genes in the HepG2 cells were examined by reverse transcription polymerase chain reaction. ResultsTreatment of HepG2 cells with 100 μg/mL aqueous or ethanol extract significantly reduced lipid accumulation and the level of triglyceride in the cells. Furthermore, pre-treatment of HepG2 cells with 100 μg/mL aqueous extract effectively down-regulated the mRNA levels of fatty acid synthase, 3-hydroxy-3-methylglutaryl CoA reductase and glycerol-3-phosphate acyl transferase, whereas 150 μg/mL ethanol extract only down-regulated the mRNA level of 3-hydroxy-3-methylglutaryl CoA reductase. ConclusionThese results provided support for the potential preventive effect of Camellia euphlebia flower on nonalcoholic fatty liver disease and could partly explain the basis of using Camellia euphlebia for the treatment of fatty liver.

The metabolomics responses of Chinese mitten-hand crab (Eriocheir sinensis) to different dietary oils

Nutritional values of animal diets depend on the type of oil in the feed ingredient, but the underlying metabolic mechanisms of dietary oil in animal feed have not been thoroughly studied in aquatic animals, especially in crustaceans. In the present study, we conducted assays on GC–MS-based metabolomics and nutritional parameters to investigate the metabolic mechanisms between juvenile Chinese mitten crabs (Eriocheir sinensis) fed olive oil containing 69% oleic acid (OA) and perilla oil containing 56% linolenic acid (LNA). Crab fed OA displayed faster growth, lower concentrations of hepatic glycogen, triglycerides and peroxidation products than those fed LNA. In the metabolomics assay, among 222 peaks isolated, 69 peaks were identified in serum. Among 13 significantly different metabolites between OA and LNA groups, six metabolites related to glycolysis and TCA (tricarboxylic acid) cycle (pyruvate, succinic acid, lactose, malic acid, D-glyceric acid and threitol), methionine, 2-keto-isovaleric acid (intermediate for valine and leucine synthesis) and 2-hydroxybutanoic acid (intermediate for glutathione synthesis) were higher in the OA group than in the LNA group. Only glutaconic acid (intermediate of ketogenic amino acids breakdown) was higher in the LNA group. This study indicates that crab in the OA group increased degradation of glucose and lipids to provide energy for growth as compared with crab in the LNA group. This is the first metabolomics study to identify the key pathways and crucial metabolites as biomarkers to differentiate the metabolic mechanisms of crustaceans fed contrasting dietary oils.

Characteristic analysis of Fe3O4 nanoparticles modified by oleic acid and undecylenic acid

ABSTRACTCarbonization of magnetic polymer microspheres is one of the methods for the preparation of magnetic carbon materials. Fe3O4 magnetic particle characteristics considerably influence the magnetic content and size distribution of magnetic polymer microspheres. The characteristics of Fe3O4 nanoparticles modified by oleic acid (OA) and undecylenic acid (UA) were analyzed by X-ray diffraction, Fourier transform infrared, scanning electron microscopy, dynamic laser light scattering, thermogravimetry/differential thermogravimetry, vibrating sample magnetometer, and water contact angle. Fe3O4 nanoparticles modified by OA and UA are nearly spherical and exhibit superparamagnetism. Fe3O4 particle size and saturation magnetization are slightly influenced by the OA and UA composition. OA and UA both are chemically adsorbed onto Fe3O4 as bidentate chelates. OA shows easier adsorption onto Fe3O4 than UA. OA groups have an expanded arrangement on OA@Fe3O4, whereas UA groups have a condensed arrangement on UA@Fe3O4. Particle lipophilicity decreases and particle clustering increases with decreasing OA content and increasing UA content on OA-UA@Fe3O4 nanoparticles.

Altered microRNA-9 Expression Level is Directly Correlated with Pathogenesis of Nonalcoholic Fatty Liver Disease by Targeting Onecut2 and SIRT1.

BackgroundMicroRNA-9 (miR-9) was detected in nonalcoholic fatty liver disease (NAFLD) patients to understand the role of miR-9 in NAFLD development.Material/MethodsBetween February 2014 and February 2015, 105 cases of NAFLD were recruited and confirmed by liver biopsy pathology, including patients with mild NAFLD (n=58) and moderate-severe NAFLD (n=47); nonalcoholic steatohepatitis (NASH) (n=53) and non-NASH (n=52); and 50 healthy participants were regarded as the healthy control group. MiR-9 expression was measured by qRT-PCR. For in vitro experiments, L-02 normal liver cells were divided into normal control group (cultured with original culture medium), dimethyl sulfoxide (DMSO) group (cultured with DMSO) and oleic acid group (cultured with oleic acid to induce fatty change), and MTT assay was used to measure the effect of different oleic acid concentrations on cell proliferation. Nile red staining was used to detect intracellular accumulation of lipid droplets. Further, synthetic miR-9 mimic and its control and miR-9 inhibitors and its control were independently transfected into L-02 cells.ResultsMiR-9 levels in the mild NAFLD group and moderate-severe NAFLD group were significantly higher than in the healthy control group (both P<0.05). Mean fluorescence intensity of lipid droplets increased with the duration of induction, and were dramatically higher in oleate-treated L-02 cells; intracellular triglyceride (TG) content was also higher. miR-9 levels significantly increased following oleate induction. Importantly, miR-9 levels were significantly elevated upon miR-9 mimic transfection. Conversely, miR-9 level was lowered with miR-9 inhibitors transfection. Additionally, Onecut2 and SIRT1 were identified as miR-9 targets.ConclusionsA positive relationship between miR-9 and steatosis was established with our results that miR-9 mimic transfection decreased intracellular lipid content. Finally, we identified 2 miR-9 targets, Onecut2 and SIRT1, which may be crucial players in NAFLD development.

Influence of Oleic Acid on Rumen Fermentation and Fatty Acid Formation In Vitro.

A series of batch cultures were conducted to investigate the effects of oleic acid (OA) on in vitro ruminal dry matter degradability (IVDMD), gas production, methane (CH4) and hydrogen (H2) production, and proportion of fatty acids. Rumen fluid was collected from fistulated goats, diluted with incubation buffer, and then incubated with 500 mg Leymus chinensis meal supplemented with different amounts of OA (0, 20, 40, and 60 mg for the CON, OA20, OA40 and OA60 groups, respectively). Incubation was carried out anaerobically at 39°C for 48 h, and the samples were taken at 12, 24 and 48 h and subjected to laboratory analysis. Supplementation of OA decreased IVDMD, the cumulative gas production, theoretical maximum of gas production and CH4 production, but increased H2 production. However, no effect was observed on any parameters of rumen fermentation (pH, ammonia, production of acetate, propionate and butyrate and total volatile fatty acid production). The concentrations of some beneficial fatty acids, such as cis monounsaturated fatty acids and conjugated linoleic acid (CLA) were higher (P < 0.05) from OA groups than those from the control group at 12 h incubation. In summary, these results suggest that the OA supplementation in diet can reduce methane production and increase the amount of some beneficial fatty acids in vitro.

Dual hit lipopolysaccharide & oleic acid combination induced rat model of acute lung injury/acute respiratory distress syndrome.

Background & objectives:Despite advances in therapy and overall medical care, acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) management remains a problem. Hence the objective of this study was to develop a rat model that mimics human ALI/ARDS.Methods:Four groups of Wistar rats, 48 per group were treated with (i) intratracheal (IT) lipopolysaccharide (LPS) (5 mg/kg) dissolved in normal saline (NS), (ii) intravenous (iv) oleic acid (OA) (250 μl/kg) suspension in bovine serum albumin (BSA), (iii) dual hit: IT LPS (2 mg/kg) dissolved in NS and iv OA (100 μl/kg) and (iv) control group: IT NS and iv BSA. From each group at set periods of time various investigations like chest X-rays, respiratory rate (RR), tidal volume (TV), total cell count, differential cell count, total protein count and cytokine levels in bronchoalveolar lavage fluid (BALF), lung wet/dry weight ratio and histopathological examination were done.Results:It was noted that the respiratory rate, and tumour necrosis factor-α (TNF-α) levels were significantly higher at 4 h in the dual hit group as compared to LPS, OA and control groups. Interleukin-6 (IL-6) levels were significantly higher in the dual hit group as compared to LPS at 8 and 24 h, OA at 8 h and control (at all time intervals) group. IL-1β levels were significantly higher in LPS and dual hit groups at all time intervals, but not in OA and control groups. The injury induced in dual hit group was earlier and more sustained as compared to LPS and OA alone.Interpretation & conclusions:The lung pathology and changes in respiration functions produced by the dual hit model were closer to the diagnostic criteria of ALI/ARDS in terms of clinical manifestations and pulmonary injury and the injury persisted longer as compared to LPS and OA single hit model. Therefore, the ARDS model produced by the dual hit method was closer to the diagnostic criteria of ARDS in terms of clinical manifestations and pulmonary injury.

Vagal efferent stimulation protects against Mesobuthus tamulus venom-induced acute respiratory distress syndrome in rats

Mesobuthus tamulus (MBT) venom and oleic acid (OA) have been shown to produce acute respiratory distress syndrome (ARDS) involving different mechanisms. The role of vagally mediated anti-inflammatory pathway in ARDS is poorly understood. Therefore, the effects of vagal efferent stimulation on these two models of ARDS were examined. Experiments were performed on anesthetized adult rats. Parameters like ventilatory changes (respiratory frequency and minute ventilation), hypoxemic status (PaO2/FiO2 ratio; P/F ratio), survival time, pulmonary water content and histopathological evidences of lung injury were determined to assess the severity of ARDS. In addition, heart rate (HR) and mean arterial pressure (MAP) were monitored. Injection of OA/MBT venom produced respiratory alterations, hypoxemia, pulmonary edema and histopathological changes demonstrating the development of ARDS. In both the groups, animals died around 60 min. Tachypnea and hyperventilation were seen after OA while bradypnea and hypoventilation were seen after MBT venom. Pulmonary edema was absent in vagotomised animals in MBT venom group but not in OA group. Further, electrical stimulation of the cut peripheral ends of vagii prolonged the survival time and attenuated all the parameters of MBT venom-induced ARDS significantly. In case of OA, there was improvement in histopathological changes but the survival time of animals was not prolonged. Stimulation of α7-nicotinic receptors (by pretreatment with GTS-21) exacerbated OA as well as MBT venom-induced ARDS. The present results indicate that vagal efferent stimulation protects against MBT venom-induced ARDS.

Clinical significance of vascular endothelial growth factor levels in bronchoalveolar lavage fluid of patients with acute lung injury

Objective To test the levels of vascular endothelial growth factor (VEGF) in bronchoalveolar lavage fluid (BALF) during acute lung injury (ALI)/ acute respiratory distress syndome (ARDS) after the interference with anticoagulation and fibrinolytic in an attempt to find a possible treatment plan for ALI/ARDS. Methods Oleic acid-induced ALI models were used in this study. A total of 96 rats were randomly assigned oleic acid group, salvianolate group, urokinase group, and saline group according to the random number table, with 24 animals per group. In oleic acid group, oleic acid (0.2 ml/kg, caudally) was administered alone. In addition to the oleic acid, the rats in salvianolate and urokinase groups received caudal injection with 10 ml/kg of 0.06% salvianolate and 5, 000 U/kg of urokinase respectively. In saline group, saline solution with an equal volume of oleic acid was caudally administered alone. Effect of the early use of salvianolate and urokinase on levels of VEGF in BALF was measured. Clinically, patients with ALI/ARDS composing survivor and death groups were enrolled and assessed by acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and multiple organ dysfunction syndrome (MODS) score. Besides, healthy population was included as the controls to comparatively analyze the levels of VEGF in BALF in ALI/ARDS group. Results Of the rat models of ALI, early use of salvianolate or urokinase was associated with elevated level of VEGF in BALF(P<0.05), which subsequently attenuated the lung injury. In the clinical research, it was found that the level of VEGF in BALF in ALI/ARDS group was much lower than that in control group [(45.9±5.7)pg/ml vs (60.6±4.5)pg/ml] (P<0.01). The level of VEGF in BALF was negatively correlated with APACHEⅡor MODS scores(r=-0.542, -0.659, P<0.01). Moreover, patients in death group appeared lower VEGF level in BALF than those in survivor group(t=2.68, P<0.05). Conclusion VEGF level in BALF can be taken as an indicator of patient condition and outcome. Early use of anticoagulation or thrombolytic drugs may play a positive role on patients with ALI/ARDS. Key words: Acute lung injury; Vascular endothelial growth factors; Salvia miltiorrhiza; Urokinase

Physicochemical and Antioxidant Characteristics of Safflower Seed Oil

The physicochemical characteristics of seed and oil of six varieties/lines of safflower cultivated in Iran and comparison of type and amount of oil and their classification to oleic (Saffola) or linoleic acid group were investigated. Varieties/lines of Iranian safflower including Goldasht (IL.III, a spring variety), Mahali Isfehan, Isfehan 14, Isfehan 28 (a spring line), Padide (LRV, a fall variety) and Sina (PI, a day land variety) were provided and characteristics of safflower seeds (oil, protein, ash, moisture and fiber contents) and their extracted oil (iodine, saponification and acid values, fatty acid profile, α-, β-, γ- and δ-tocopherol, tocotrienols and β-carotene) were studied. Results indicated that the studied samples were only linoleic safflower (not oleic safflower). Iranian safflower varieties/lines have the highest amount of ω -6 (71.99 - 75.78) in comparison with other oil sources which is a valuable and essential fatty acid. Goldasht and Mahali Isfehan lines as a natural antioxidant (α-tocopherol: 346.25 – 376.83%; β-tocopherol: 54.29 – 52.89%; β-carotene: 8.53 – 8.1%, respectively) were mixed at two levels of 2 and 5% with soybean oil (without any antioxidant) and their shelf lives were determined by peroxide, thiobarbituric acid (TBA) and Rancimat indices at 0, 5, 10 and 15 days and were compared with control. Statistical results did not show any antioxidant effect in shelf life of soybean oil at more days. This means that the safflower antioxidant compounds were not migrated completely to its oil. Keywords: Antioxidant characteristics, physicochemical characteristics, safflower, safflower oil, varieties/lines.

Regulatory effects of hydrogen sulfide on alveolar epithelial cell endoplasmic reticulum stress in rats with acute lung injury

The present study was undertaken to examine the regulatory effect of hydrogen sulfide (H2S) on endoplasmic reticulum stress in alveolar epithelial cells of rats with acute lung injury (ALI) induced by oleic acid (OA). Seventy-two male Sprague Dawley (SD) rats were divided into control group, oleic acid-induced ALI group (OA group), oleic acid-induced ALI with sodium hydrosulfide (NaHS) pretreatment group (OA+NaHS group), and sodium hydrosulfide treatment group (NaHS group). Rats of each group were further subdivided into 3 subgroups. Index of quantitative assessment of histological lung injury (IQA), wet/dry weight ratio (W/D) and H2S level of lung tissues were measured. The expressions of endoplasmic reticulum stress markers including glucose-regulated protein 78 (GRP78) and α-subunit of eukaryotic translation initiation factor-2 (elF2α) in lung tissues were measured by immunohistochemical staining and Western blotting. The IQA score and W/D ratio of lung tissues at the three time points significantly increased in rats injected with OA, but significantly decreased in other rats injected with OA and NaHS. The level of H2S in lung tissue at the three time points significantly decreased in rats injected with OA, but significantly increased in other rats injected with both OA and NaHS. GRP78 and elF2α decreased in rats injected with OA, but increased in other rats injected with both OA and NaHS, especially at 4-hour and 6-hour time points. The results suggested that H2S could promote alveolar epithelial cell endoplasmic reticulum stress in rats with ALI.

Study of synergistic effect of free fatty acid and iron on the establishment of nonalcoholic fatty liver disease model

To establish nonalcoholic fatty liver disease (NAFLD) model induced by free fatty acid (FFA) and iron, and to explore the synergistic effect of FFA and Fe(2+) on the pathogenesis of NAFLD and mechanisms. Human liver carcinoma cell HepG2 was respectively treated with 0.250, 0.500, 1.000 mmol/L oleic acid, 0.500 mmol/L oleic acid+0.125 mmol/L Fe(2+), 0.500 mmol/L oleic acid+0.250 mmol/L Fe(2+), and 0.500 mmol/L oleic acid+0.500 mmol/L Fe(2+). Human liver carcinoma cell HepG2 was normally cultured in the control group. Lipid accumulation of cells were observed by oil red O staining and the determination of the triglyceride (TG) contents by GPO-PAP, then the expression of key genes involved in fatty acid β-oxidation (fatty acyl CoA synthetase-1 (ACSL-1), carnitine acyl transferase 1 (CPT-1a), fatty acid synthetase (FAS)) was determined using RT-PCR. The differences of TG content and ACSL-1, CPT-1a, FAS, mRNA relative value were analyzed among different groups. The results of oil red O staining indicated that the contents of lipid droplets were obviously elevated with the increase of Fe(2+) concentration in human liver carcinoma cell HepG2 treated with 0.500 mmol/L oleic acid and different concentrations of Fe(2+). The TG contents of HepG2 cell in control group, 0.250, 0.500, 1.000 mmol/L oleic acid groups, 0.500 mmol/L oleic acid+0.125 mmol/L Fe(2+) group, 0.500 mmol/L oleic acid+0.250 mmol/L Fe(2+) group, 0.500 mmol/L oleic acid+0.500 mmol/L Fe(2+) group respectively were (90.0 ± 1.6), (131.7 ± 5.4), (153.7 ± 3.0), (254.1 ± 4.0), (164.5 ± 6.0), (180.1 ± 7.7), (235.6 ± 4.5) nmol/mg (F = 396.00, P < 0.05). The expression levels of ACSL-1 mRNA in 0.500 mmol/L oleic acid group, 0.500 mmol/L oleic acid+0.125 mmol/L Fe(2+) group, 0.500 mmol/L oleic acid +0.250 mmol/L Fe(2+) group, 0.500 mmol/L oleic acid +0.500 mmol/L Fe(2+) group respectively were (0.94 ± 0.02), (0.89 ± 0.04), (0.85 ± 0.02), (0.74 ± 0.04) (F = 50.00, P < 0.05); the mRNA levels of CPT-1a were (0.89 ± 0.03), (0.79 ± 0.05), (0.67 ± 0.04), (0.51 ± 0.05) (F = 79.00, P < 0.05); the mRNA levels of FAS were (1.31 ± 0.05) , (1.44 ± 0.03), (1.51 ± 0.05), (1.56 ± 0.06 ) (F = 79.70, P < 0.05). The NAFLD liver cell model could be established by oleic acid and Fe(2+) in HepG2 cells. FFA and iron might be involved in the pathogenesis of NAFLD through the intervention of fatty acid β-oxidation.

Xuebijing injection improves the respiratory function in rabbits with oleic acid-induced acute lung injury by inhibiting IL-6 expression and promoting IL-10 expression at the protein and mRNA levels.

Xuebijing injection is a complex herbal medicine, and clinical and experimental studies have shown that it has a significant effect on acute respiratory distress syndrome and multiple organ dysfunction syndrome. However, the majority of studies regarding Xuebijing injection have focused on serum inflammatory factors, and few studies have been carried out from the perspective of the protein and mRNA expression of inflammatory cytokines. In this study, 60 healthy rabbits of mixed gender were randomly assigned to a normal control group (CG), oleic acid group (model group; MG) and oleic acid + Xuebijing injection group (treatment group; TG). Rabbits of the CG were treated with normal saline through the ear vein, rabbits of the MG were injected with oleic acid (0.4 ml/kg) and rabbits of the TG received 0.4 ml/kg oleic acid + 10 ml/kg Xuebijing injection. Blood samples were collected from the common carotid artery of all rabbits of all groups 1 h after the ear vein was injected with the corresponding reagent, and was used to measure the arterial partial pressure of oxygen (PaO2) and of carbon dioxide (PaCO2). The activity of myeloperoxidase (MPO) was tested, and the protein and mRNA expression levels of interleukin (IL)-6 and IL-10 were determined. Rabbits of the MG exhibited evident respiratory dysfunction (PaO2 and PaCO2 were low), histopathological lung damage and overactive inflammatory responses (the expression of the proinflammatory cytokine IL-6 and the anti-inflammatory cytokine IL-10 was increased at the protein and mRNA levels). Following the administration of the Xuebijing injection, the inflammatory response of the rabbits was significantly reduced. Xuebijing injection raised PaO2 and PaCO2, weakened the activity of MPO in the lung tissue, downregulated the expression of the proinflammatory cytokine IL-6 and further increased the expression of the anti-inflammatory cytokine IL-10. These results demonstrated that Xuebijing injection improved the respiratory function of rabbits with acute oleic acid-induced lung injury by inhibiting IL-6 expression and promoting IL-10 expression.

Nanoparticle lattices and surfaces

Nanomaterials The challenge of resolving the details of the surfaces or assemblies of colloidal semiconductor nanoparticles can be overcome if several characterization methods are used (see the Perspective by Boles and Talapin). Boneschanscher et al. examined honeycomb superlattices of lead selenide nanocrystals formed by the bonding of crystal faces using several methods, including high-resolution electron microscopy and tomography. The structure had octahedral symmetry with the nanocrystals distorted through “necking”: the expansion of the contact points between the nanocrystals. Zherebetskyy et al. used a combination of theoretical calculations and spectroscopic methods to study the surface layer of lead sulfide nanocrystals synthesized in water. In addition to the oleic acid groups that capped the nanocrystals, hydroxyl groups were present as well. Science , this issue p. [1377][1], p. [1380][2]; see also p. [1340][3] [1]: /lookup/doi/10.1126/science.1252642 [2]: /lookup/doi/10.1126/science.1252727 [3]: /lookup/doi/10.1126/science.1256197

Long-range orientation and atomic attachment of nanocrystals in 2D honeycomb superlattices.

Oriented attachment of synthetic semiconductor nanocrystals is emerging as a route for obtaining new semiconductors that can have Dirac-type electronic bands such as graphene, but also strong spin-orbit coupling. The two-dimensional (2D) assembly geometry will require both atomic coherence and long-range periodicity of the superlattices. We show how the interfacial self-assembly and oriented attachment of nanocrystals results in 2D metal chalcogenide semiconductors with a honeycomb superlattice. We present an extensive atomic and nanoscale characterization of these systems using direct imaging and wave scattering methods. The honeycomb superlattices are atomically coherent and have an octahedral symmetry that is buckled; the nanocrystals occupy two parallel planes. Considerable necking and large-scale atomic motion occurred during the attachment process.

Hydroxylation of the surface of PbS nanocrystals passivated with oleic acid.

Controlling the structure of colloidal nanocrystals (NCs) is key to the generation of their complex functionality. This requires an understanding of the NC surface at the atomic level. The structure of colloidal PbS NCs passivated with oleic acid has been studied theoretically and experimentally. We show the existence of surface OH(-) groups, which play a key role in stabilizing the PbS(111) facets, consistent with x-ray photoelectron spectroscopy as well as other spectroscopic and chemical experiments. The role of water in the synthesis process is also revealed. Our model, along with existing observations of NC surface termination and passivation by ligands, helps to explain and predict the properties of NCs and their assemblies.

MicroRNAs mediate oleic acid-induced acute lung injury in rats using an alternative injury mechanism.

Intravenous (IV) infusion of oleic acid (OA) distributes OA microemboli in the pulmonary capillaries, which results in severe vascular congestion, hemorrhage vascular congestion, interstitial edema, intravascular coagulation and bleeding. The immune response to acute lung injury (ALI) is known to be associated with rapid and widespread changes in microRNA (miRNA) expression in the lung. The present study of a model of rat lung injury aimed to investigate how the lung miRNA profile changes to mediate ALI. For the induction of ALI, OA (200µl/kg) suspended in 20% ethyl alcohol was injected through the tail vein for 20min. Lung tissue samples were acquired at 3, 6 and 24h, and miRNA microarray and quantitative polymerase chain reaction were performed using these samples. The activation of phosphatase and tensin homolog (PTEN), protein kinase B (Akt), extracellular signal-regulated kinases (ERK) and c-Jun N-terminal kinases (JNK) were analyzed by western blot analysis. There were 75 miRNAs that demonstrated >1.5‑fold changes in expression levels. miR-101a was highly upregulated at 3h. miR-21 was upregulated in the OA group throughout the 24h following OA challenge. miR-1 was the most downregulated miRNA at 24h. In order to examine the expression levels of PTEN and Akt as targets of miR-21, western blot analysis was performed. At 3h, the levels of PTEN were attenuated in the OA group as compared with those in the control group; however, p-Akt/Akt levels were increased at 3h for the OA group. PTEN and p-Akt/Akt were significantly higher in the OA group at 3h and were rapidly decreased at 6h. The immunohistochemical stain of α-smooth muscle actin in the bronchial and alveolar wall increased 24h after OA‑induced ALI. These results indicated that the profile of miRNAs dynamically changed throughout the OA-induced ALI process, and mitogen-activated protein kinase activation, PTEN/Akt pathway alteration and smooth muscle actin activation were observed in this ALI model.

Stimulating effect of essential oils from Ligusticum chuanxiong Hort on HaCaT cells and skin

Objective To investigate the skin stimulation effect of essential oils from Ligusticum chuanxiong Hort. Methods HaCaT cells were cultured in vitro.Using MTT and ELISA methods,we examined the effect of the essential oil (different concentrations) on HaCaT cell proliferation and prostaglandin E_2(PGE_2) levels in culture supernatants of HaCaT cells,and the results were compared with those of oleic acid,a classic permeation enhancer.The cumulative skin stimulation effect was determined by visual scoring in guinea pigs and the histological changes were determined by light microscopy.Results The HaCaT cell viabilities of the 0.005%,0.015%,and 0.025%essential oil groups were 1.79-,1.65-,and 1.48-fold that of the 0.005%oleic acid group,respectively,and there was no significant difference between the 0.05%essential oil group and 0.005%oleic acid group.The 0.005%,0.015%,0.025%,and 0.05%essential oil influenced the supernatant PGE_2 levels by (0.99±0.08)%,(1.63±0.09)%,(0.98±0.09)%,and(0.04±0.01)%,respectively,which were all significantly lower than the influence of 0.005%oleic acid([4.23±0.68]%,P0.05).Only slight erythema was observed after continuous administration the essential oil(different concentrations) for 7 days,with no edema or skin uplift,and the erythema caused by 15%essential oil was lower than that caused by 5%oleic acid.Only 15%essential oil exhibited the mechanical injury of the stratum corneum.And 5%oleic acid group showed stripped and lost stratum corneum over large areas. Conclusion Chuanxiong oil is less cytotoxic and less stimulating to the skin compared with oleic acid,and may become an excellent skin permeation enhancer.

Expression of aquaporin 3 and aquaporin 9 is regulated by oleic acid through the PI3K/Akt and p38 MAPK signaling pathways

To study the effect of oleic acid (OA) on expression of aquaglyceroporin genes, AQP3 and AQP9, in hepatocyte steatosis and to investigate the underlying molecular mechanisms using an in vitro system. HepG2 cells were treated with OA at different concentration to establish in vitro models of nonalcoholic hepatocyte steatosis. The corresponding extents of hepatic steatosis modeling were assessed by oil red O staining and optical density (OD) measurements of the intracellular fat content. The model lines were then treated with inhibitors of the PI3K/Akt and p38 MAPK signaling pathway factors and effects on AQP3/9 expression was measured by real time RT-PCR and western blotting. The fat concentration, indicative of hepatic steatosis, increased in conjunction with increased concentrations of OA (0 less than 250 less than 500 mumol/L). OA exposure also down-regulated AQP3 mRNA and up-regulated AQP9 mRNA levels in a concentration-dependent manner. The most robust changes in expression occurred in response to the 500 mumol/L concentration of OA for both AQP3 (0.47+/-0.18; t = 4.5450, P less than 0.05) and AQP9 (1.57+/-0.21; t = 3.0306, P less than 0.05). Treatment with OA + PI3K pathway inhibitor (LY294004) significantly decreased AQP9 mRNA expression (4.55+/-0.62) as compared to the control group (1.00+/-0.10; t = 9.7909, P less than 0.01), that 500 mumol/L OA group (2.43+/-0.53; t = 4.5018, P less than 0.05), and the LY294002 group (1.90+/-0.16; t = 7.1683, P less than 0.01). Treatment with p38 MAPK pathway inhibitor (SB230580) significantly increased the OA-suppressed level of AQP3 mRNA to the level detected in the control group (1.27+/-0.11; t = 5.7455, P less than 0.01) and decreased the OA-stimulated AQP9 mRNA (0.38+/-0.09; t = 6.5727, P less than 0.01). No significant changes in mRNA expression of AQP3/9 were observed with inhibition of the ERK1/2 and JNK signal transduction pathways. The OA-induced changes in protein expression levels of AQR3 and AQP9 followed a similar trend of the genes. Finally, OA suppressed the level of phosphorylated Akt (from 0.21+/-0.02 to 0.13+/-0.03; t = 3.8431, P less than 0.05) but elevated the level of phosphorylated p38 (from 0.58+/-0.06 to 1.02+/-0.10; t = 12.5289, P less than 0.01). Again, OA treatment produced no significant affect on ERK1/2 and JNK phosphorylation. OA down-regulates AQP3 expression by stimulating the p38 MAPK signaling pathway, and up-regulates the AQP9 by blocking the PI3K/Akt pathway and activating the p38 MAPK signaling pathway.

Oleic Acid Inhibits the KATP Channel Subunit Kir6.1 and the KATP Current in Human Umbilical Artery Smooth Muscle Cells

BackgroundThe objective of the present study was to determine the effect of various concentrations of oleic acid (OA) on KATP channel expression and the potential relationship to exogenous nitrogen monoxide and protein kinase C levels. MethodsHuman umbilical artery smooth muscle cells (HUASMCs), between the 7th and 10th passages, were divided into control group, OA group (final OA concentration of 0, 50, 100 or 200 μmol/L), nitric oxide (NO) intervention group, protein kinase C inhibitor group or GF-109203X (GFX) intervention group. Western immunoblotting was used to detect the protein expression of the KATP channel subunit Kir6.1. Also, quantitative real-time polymerase chain reaction analysis to determine Kir6.1 messenger RNA levels and whole-cell patch clamping to measure KATP currents were performed. ResultsThe results suggested that OA inhibited Kir6.1 protein and messenger RNA expression in HUASMCs. Under a high concentration of potassium (140 mmol/L), 100 μmol/L OA significantly reduced ATP-sensitive potassium current density, whereas a low extracellular concentration of potassium (5.4 mmol/L) did not influence KATP density. Pretreatment with either exogenous NO or GFX weakened the OA-induced inhibition of KATP in HUASMCs. ConclusionsThe study demonstrated that OA inhibited Kir6.1, a KATP channel subunit, in HUASMCs, and indirectly inhibited the KATP current. In addition, the results indicated that NO and/or GFX partially reversed OA inhibition in HUASMCs.

Polyunsaturated fatty acid relatively decreases cholesterol content in THP-1 macrophage-derived foam cell: partly correlates with expression profile of CIDE and PAT members

BackgroundPolyunsaturated fatty acids (PUFAs) have positive effect on the regulation of plasma lipids. But the mechanism for them to modulate lipid homeostasis in macrophage is still unclear. In this study, we employed PUFA to pretreat macrophages and evaluated the variations of lipid droplet (LD) content, lipid composition, and expressions of LD-associated genes in macrophage-derived foam cells.MethodTHP-1-derived macrophages or human peripheral blood monocyte-derived macrophages were pre-treated with four non-esterified fatty acids (NEFAs) separately: saturated fatty acid (SFA)-palmitic acid (PA), monounsaturated fatty acids (MUFAs)-oleic acid (OA), PUFAs-linoleic acid (LA) and eicosapentaenoic acid (EPA). Intracellular lipid content and cholesterol efflux were analyzed in THP-1 macrophage-derived foam cells. Related gene expressions were detected by quantitative real-time PCR.ResultsPUFA pre-treatment reduced cholesterol content in foam cells and increased cholesterol efflux to lipid-free apoAI in conditioned medium compared with PA or OA group. Cell death-inducing DFF45 like effector (CIDE) and Perilipin-Adipophilin-TIP47 (PAT) family members, as LD-associated proteins, showed specific gene expression profiles after PUFA pre-treatment. These results may help to explain the process of lipid metabolism within foam cells.ConclusionPUFA (LA or EPA) had a potential protective effect against cholesterol accumulation. The specific expressions of CIDE and PAT genes may provide clues to explore the protective mechanism of PUFA in foam cells.