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Abstract Background: Elevated serum-free fatty acid (FFA) levels induce insulin resistance (IR) or a protective mechanism to IR development in humans; it depends on FFA type. Objetive: This study explores the effects of oleic (OLA - unsatured) and palmitic (PAM - saturated) fatty acids on insulin action in mature adipocytes effect. Methods: Cells were incubated 18 h with or without OLA and PAM at 250 μM, and 500 μM. After the culture period, were measured: adipocyte viability, size, fatty acids mobilisation, insulin signalling proteins, and glucose uptake. Results: Adipocytes exhibited optimal viability tolerances regardless of the kinds of fatty acids used for treatment. However, adipocytes were hypertrophic after OLA and PAM stimuli. Additionally, lipogenesis (lipid synthesis), and lipolysis (lipid breakdown) were significantly increased by treatment with OLA, or PAM (500 μM) compared to control. Moreover, OLA results showed that there was no significant reduction in signalling cascades, except for a downstream proinflammatory response. Instead, PAM hypertrophic adipocytes were insulin resistant with alteration of proinflammatory and stress markers. Conclusions: Current findings suggest that PAM induces insulin resistance, mitochondrial and reticulum stress on fat cells compared to those treated with OLA that, protects adipocytes to all those alterations.
Resumen Introducción: Los niveles elevados de ácidos grasos libres (AGL) en suero inducen resistencia a insulina (RI) o un mecanismo de protección del desarrollo de RI en humanos, esto depende del tipo de AGL. Objetivo: Este estudio explora los efectos de los ácidos grasos oleico (insaturados - OLA) y palmítico (saturados - PAM) sobre la insulina en adipocitos maduros. Métodos: Las células se incubaron 18 h con o sin OLA y PAM a 250 μM y 500 μM. Después del período de cultivo, se evaluó en adipocitos: viabilidad, tamaño, movilización de ácidos grasos, proteínas de señalización de insulina y absorción de glucosa. Resultados: Los adipocitos mostraron viabilidad óptima independientemente de los tipos de ácidos grasos utilizados en el tratamiento. Los adipocitos eran hipertróficos tras estimulo con OLA y PAM. La lipogénesis (síntesis de lípidos) y la lipólisis (degradación de lípidos) aumentaron significativamente con el tratamiento con OLA o PAM (500 μM) en comparación con el control. En los resultados de OLA no se evidenció una reducción significativa en las cascadas de señalización de insulina, a excepción de una respuesta proinflamatoria posterior. En cambio, los adipocitos hipertróficos tratados con PAM presentaron resistencia a la insulina y alteración de los marcadores proinflamatorios y de estrés. Conclusiones: Nuestros hallazgos sugieren que PAM induce resistencia a la insulina, estrés mitocondrial y del retículo en las células grasas en comparación con aquellos tratados con OLA, AGL que, en cambio, protegen a los adipocitos de todas esas alteraciones.
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Insulin Resistance , Adipocytes , Palmitic Acid , Oleic Acid , Fatty AcidsABSTRACT
@#To investigate the regulatory effects of insulin and oleic acid on serum metabolites in colon cancer, subcutaneous transplantation tumor model of colon carcinoma cell HCT116 was established. Nude mice were randomly divided into 4 groups: control (CON, vehicle); insulin treatment (INS, sc, 2.5 U/kg); oleic acid treatment (OA, ig, 2.0 g/kg); and insulin (sc, 2.5 U/kg) plus oleic acid (ig, 2.0 g/kg) treatment (IO). Non-target metabolomic analysis on the blood samples was performed by GC/MS and LC-IT-TOF/MS. Data pre-processing, including peaking, spectral deconvolution and peak alignment, was performed before data were imported to SIMCA-P for multivariate statistical analysis. Results showed that body weight of individuals in IO group was the lowest, but the tumor weight was the heaviest. Metabolic profiles of IO group were also different compared with the CON group, and the contributing metabolites were urea, arabinose, cholesterol, L-acetylcarnitine and sphingosine. There was no significant difference between OA or INS and CON. This study showed that the combination of insulin and oleic acid promoted colon cancer deterioration and caused metabolic disturbance in blood.Our study may provide theoretical foundation for the discovery of colon cancer biomarker and its early diagnosis.
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OBJECTIVE: To investigate the effects and possible mechanism of trans-oleic acid (9t-C18:1) on proliferation inhibition and induction apoptosis in H9c2 cardiomyocyte. METHODS: H9c2 rat cardiomyocytes were cultured in vitro. High, medium and low (600, 300, 150 μmol•L-1) dose of 9t-C18:1 groups and the negative control (NC) group were administered to H9c2 cardiomyocytes. The effect of 9t-C18:1 on cell proliferation was tested using cell counting kit-8 (CCK-8) assay. Morphological changes of cells were observed by AO-EB staining. Intracellular reactive oxygen species (ROS) and apoptosis were detected by flow cytometry. The expression of Bcl-2 and Bax genes was detected by quantitative real time- polymerase chain reaction (QRT-PCR). The expression of Bcl-2 and Bax protein was determined by flow cytometry after immunofluorescence staining. RESULTS: The typical morphological characteristics of apoptosis were observed by fluorescence microscope. The result of CCK-8 assay indicated that 9t-C18:1 have an certainly inhibitory effect on the proliferation of H9c2 cells. ROS level and apoptosis rate were significantly increased. Bcl-2 gene and protein expression were down-regulated, and Bax gene and protein expression were up-regulated, compared with NC group(P<0.05, P<0.01). CONCLUSION: 9t-C18:1 can inhibit the proliferation and induce the apoptosis of H9c2 cardiomyocyte, and its mechanism may be related to promoting the mitochondrial apoptotic pathway.
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OBJECTIVE: To establish a mini-column centrifugation method to determine the encapsulation efficiency (EE) of novel solid lipid nanoparticles containing oleic acid-CAT3 conjugates (OA-CAT3-SLN) and normal CAT3 SLN (CAT3-SLN). METHODS: Sephadex G-50 mini-columns were used to separate the encapsulated drug and free drug in the solid lipid nanoparticles (SLN) with the help of centrifugation. The boundary point of the elution between the encapsulated drug and free drug was established by the elution curve. The encapsulated drug in the SLN was eluted with 1 mL water for three times. Then 2 mL of ethanol was used to elute the separated free drug for three times. The eluted CAT3 was quantified by the verified HPLC-UV method and used to calculate the EE. The method was verified with the recovery and repeatability test. Finally, the EE of three batches of OA-CAT3-SLN and CAT3-SLN were determined. RESULTS: The mini-column centrifugation method could effectively separate the free drug from the encapsulated drug in SLN. The column recovery was (99.64±1.97)% (n=9), and the result of EE repeatability test of OA-CAT3-SLN was (83.71±0.70)% (n=5). The EE of OA-CAT3-SLN and CAT3-SLN were (86.26±2.65)% and (72.22±4.52)%, respectively (n=3). CONCLUSION: The established separation method is simple and reliable, with high recovery and good repeatability, and can distinguish different preparation processes.
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Objective: To establish a rapid qualitative analysis method for fatty acids and esters in Coicis Semen by ultra-performance liquid chromatography triple quadrupole time-of-flight mass spectrometry (UPLC-Triple-TOF-MS). Methods: Agilent ZORBAX-SB C18 (250 mm × 4.6 mm, 5.0 μm) column was used. The mobile phase was acetonitrile-isopropanol (1:1) elution, the flow rate was 1 mL/min, the detection wavelength was 210 nm, the column temperature was 30 ℃, and the injection quantity was 10 μL. Electrospray ion source positive ion mode was adopted, and the scanning range was m/z 100-1 500. The sample data were collected by full scanning mode, and the fatty acid chemical composition of Coicis Semen was quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results: A total of 29 kinds of fatty acids and esters in Coicis Semen were detected, and the cracking rules of the compounds were analyzed. Through the mass-to-charge ratio of molecular ion peaks and fragment ions, the Scifinder and Reaxy network databases, and the literature, these compounds were different under the action of ion source by losing the structure of oleic acid, linoleic acid, palmitic acid, oxidizing oleic acid and the like. The mass-to-charge ratio of the fragment ions, and the name and structural formula of the 29 fatty acids and their ester compounds were inferred. Conclusion: The method of qualitative analysis of the fatty acids and esters of Coicis Semen established in this study is accurate, rapid and sensitive, which provides experimental basis for improving the quality control level of Coicis Semen and further elucidating the pharmacodynamic substance basis of Coicis Semen.
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Stearoyl-ACP Δ⁹ desaturase (SAD) catalyzes the synthesis of monounsaturated oleic acid or palmitoleic acid in plastids. SAD is the key enzyme to control the ratio of saturated fatty acids to unsaturated fatty acids in plant cells. In order to analyze the regulation mechanism of soybean oleic acid synthesis, soybean (Glycine max) GmSAD family members were genome-wide identified, and their conserved functional domains and physicochemical properties were also analyzed by bioinformatics tools. The spatiotemporal expression profile of each member of GmSADs was detected by qRT-PCR. The expression vectors of GmSAD5 were constructed. The enzyme activity and biological function of GmSAD5 were examined by Agrobacterium-mediated transient expression in Nicotiana tabacum leaves and genetic transformation of oleic acid-deficient yeast (Saccharomyces cerevisiae) mutant BY4389. Results show that the soybean genome contains five GmSAD family members, all encoding an enzyme protein with diiron center and two conservative histidine enrichment motifs (EENRHG and DEKRHE) specific to SAD enzymes. The active enzyme protein was predicted as a homodimer. Phylogenetic analysis indicated that five GmSADs were divided into two subgroups, which were closely related to AtSSI2 and AtSAD6, respectively. The expression profiles of GmSAD members were significantly different in soybean roots, stems, leaves, flowers, and seeds at different developmental stages. Among them, GmSAD5 expressed highly in the middle and late stages of developmental seeds, which coincided with the oil accumulation period. Transient expression of GmSAD5 in tobacco leaves increased the oleic acid and total oil content in leaf tissue by 5.56% and 2.73%, respectively, while stearic acid content was reduced by 2.46%. Functional complementation assay in defective yeast strain BY4389 demonstrated that overexpression of GmSAD5 was able to restore the synthesis of monounsaturated oleic acid, resulting in high oil accumulation. Taken together, soybean GmSAD5 has strong selectivity to stearic acid substrates and can efficiently catalyze the biosynthesis of monounsaturated oleic acid. It lays the foundation for the study of soybean seed oleic acid and total oil accumulation mechanism, providing an excellent target for genetic improvement of oil quality in soybean.
Subject(s)
Fatty Acid Desaturases , Genetics , Metabolism , Gene Expression Profiling , Oleic Acid , Phylogeny , Plant Proteins , Genetics , Seeds , Chemistry , Soybeans , Classification , GeneticsABSTRACT
AIM: To study the protective effects of ginsenoside Rg1 on HL-7702 cells injury induce by oleic acid (OA), and investigate its role in aldolase/AMPK/PINK1 signalling. METHODS: HL-7702 cells were cultured in vitro and divided into five groups: control group (C), oleic acid group (OA), OA+ginsenoside (Rg1) group, OA+compound C (CC) group, and OA+CC+Rg1 group. The viability of HL-7702 cells was determined by CCK8 assay and Hoechst staining. The apoptosis and mitochondrial membrane potentials of HL-7702 cells were measured using flow cytometry. ATP content in HL-7702 cells was observed. Western blot was used to detect the expression levels of Cleaved caspase-3, PINK1, MFN2, Aldolase and p-AMPK in HL-7702 cells. RESULTS:Compared with C group, the viability of cells in OA group was significantly decreased (P<0.05), the apoptotic rate and Cleaved caspase-3 expression were greatly increased (P<0.05), ATP level, mitochondrial membrane potentials (TMRE) and PINK1 expression were significantly decreased (P<0.05). Compared with OA group, the viability of cells in OA+Rg1 group was significantly increased (P<0.05), the apoptotic rate and Cleaved caspase-3 expression were greatly decreased (P<0.05), ATP level, mitochondrial membrane potentials (TMRE) and PINK1 expression were significantly increased (P<0.05). Compared with OA+Rg1 group, the viability of cells and p-AMPK expression level in OA+CC+Rg1 group was significantly decreased (P<0.05). Reducing the expression of aldolase in cells inhibited Rg1?s actions on PINK1 and p-AMPK and cell viability. CONCLUSION: Ginsenoside Rg1 can improve the injury of HL-7702 cells via regulating aldolase/AMPK/PINK1 signaling pathway.
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Background & objectives: Stearoyl-CoA desaturase 1 (SCD1) is a key lipogenic enzyme responsible for endogenous synthesis of monounsaturated fatty acids (MUFA) and plays a key role in various pathophysiology, including fatty liver diseases. In this experimental study the impact of vitamin A deficiency was assessed on SCD1 regulation in relation to kidney biology, under high fructose (HFr) diet-fed condition in rats. Methods: Forty male weanling (21 day old) Wistar rats were divided into four groups control, vitamin A-deficient (VAD), HFr, VAD with HFr consisting of eight rats each, except 16 for the VAD group. The groups received one of the following diets: control, VAD, HFr and VAD with HFr for 16 wk, except half of the VAD diet-fed rats were shifted to HFr diet, after eight week period. Results: Feeding of VAD diet (alone or with HFr) significantly reduced the kidney retinol (0.51, 0.44 ?g/g vs. 2.1 ?g/g; P < 0.05), while increased oleic (C18:1) and total MUFA levels (23.3, 22.2% and 27.3, 25.4% respectively vs. 14.7 and 16.6%; P < 0.05) without affecting the SCD1, both at protein and mRNA levels, when compared with HFr. Comparable, immunohistological staining for SCD1 was observed in the distal convoluted tubules. Despite an increase in MUFA, morphology, triglyceride content and markers of kidney function were not affected by VAD diet feeding. Interpretation & conclusions: Feeding of VAD diet either alone or under HFr condition increased the kidney oleic acid (C18:1) levels and thus total MUFA, which corroborated with elevated SCD1 activity index, without affecting its expression status. However, these changes did not alter the kidney morphology and function. Thus, nutrient-gene regulation in kidney biology seems to be divergent.
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In this study, magnetic nanoparticles (MNPs) coated with a combination of oleic acid and chitosan were synthesizedby ex situ and in situ coprecipitation methods. Morphology and particle size, crystal structure and crystallite size,chemical structure, and magnetic saturation were characterized by scanning electron microscope (SEM), X-raydiffraction (XRD), Fourrier transform infrared, and vibrating sample magnetometer (VSM), respectively. SEMimages showed that better spherical morphology is obtained by ex situ co-precipitation method. The XRD patternidentified that nanoparticles containing Fe3O4 and γ-Fe2O3. The particles and crystallite size of the nanoparticles tendedto decrease with increasing oleic acid to the optimum composition. Further functionalization through the chitosanaddition (crosslinked by Tripolyphosphate/sulfate) is contributed to the hydrophilicity properties of nanoparticles.Through VSM analysis, MNPs-oleic acid-chitosan showed superparamagnetic behavior with magnetic saturationreaching 32.63 emu/g. There was a linear correlation between magnetic saturation and Fe3O4 content of nanoparticles.Drug loading and drug release were carried out by using Doxorubicin. These nanoparticles showed a high drug loadingefficiency with lower chitosan composition. Loading efficiency of Doxorubicin is related to the conjugation withcarboxylic groups and hydrophobic sites from oleic acid and MNPs
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Resumen Introducción. El hígado juega un papel importante en la homeostasis lipídica, especialmente en la síntesis de ácidos grasos y triglicéridos. Una amplia variedad de modelos celulares ha sido utilizada para investigar el metabolismo lipídico hepático y para elucidar detalles específicos de los mecanismos bioquímicos del desarrollo y progresión de enfermedades relacionadas, brindando información para tratamientos que reduzcan su impacto. Los modelos celulares hepáticos poseen un alto potencial en la investigación del metabolismo de lípidos y de agentes farmacológicos o principios activos que permiten la reducción de la acumulación de lípidos. Objetivo. Comparar algunos modelos celulares hepáticos utilizados para el estudio del metabolismo lipídico, sus características y los resultados más relevantes de investigación en ellos. Materiales y métodos. Se realizó una búsqueda sistemática en bases de datos sobre los modelos celulares hepáticos de mayor uso para el estudio del metabolismo de lípidos. Resultados. Se exponen los cinco modelos celulares más utilizados para este tipo de investigaciones, destacando su origen, aplicación, ventajas y desventajas al momento de estimular el metabolismo lipídico. Conclusión. Para seleccionar el modelo celular, el investigador debe tener en cuenta cuáles son los requerimientos y el proceso que desea evidenciar, sin olvidar que los resultados obtenidos solo serán aproximaciones de lo que en realidad podría suceder a nivel del hígado como órgano.
Abstract Introduction: The liver plays an important role in lipid homeostasis, especially in the synthesis of fatty acids and triglycerides. A wide variety of cell models have been used to investigate liver lipid metabolism and to elucidate specific details of the biochemical mechanisms involved in the development and progression of related diseases, providing information for treatments that reduce their impact. Liver cell models have a high potential for the investigation of lipid metabolism, as well as pharmacological agents or active principles that allow the reduction of lipid accumulation. Objective: To compare some liver cell models used for studying lipid metabolism, their characteristics and the most relevant research results. Materials and methods: A systematic search of databases was performed on the most commonly used liver cell models for the study of lipid metabolism. Results: The five most commonly used cell models for this type of research are presented in this paper, highlighting their origin, application, advantages and disadvantages when stimulating lipid metabolism. Conclusion: In order to select a cell model, researchers should take into account the requirements and the process they wish to demonstrate, without forgetting that the results obtained will only be approximations of what could actually happen in the liver as an organ.
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Objective:To prepare magnetic solid lipid nanoparticles co-loaded quercetin and resveratrol(QR-MSLN),develop the reasonable characterization method,and investigate its inhibitory effect on transplanted hepatocarcinoma H22 tumor in mice. Method:Magnetic Fe3O4 particles coated with oleic acid(OA-Fe3O4) were synthesized and its structure was confirmed by Fourier transform infrared spectrometer(FT-IR).QR-MSLN was prepared by emulsion ultrasonic dispersion method,its morphology was examined by transmission electron microscopy,its particle size was determined by laser particle sizer.Concentration of Fe in the preparation was determined by phenanthroline spectrophotometry.The entrapment efficiency,saturation magnetization,in vitro release behavior were investigated by ultrafiltration centrifugation,vibrating sample magnetometer(VSM) and dialysis method,respectively.Mouse tumor model transplanted with hepatoma H22 ascites tumor was established and antitumor effect of QR-MSLN on H22 bearing mice were observed in the presence of an applied magnetic field. Result:Morphology of QR-MSLN was round,and black magnetic particles could be observed inside it,its particle size was (171.9±2.2) nm,the concentration of Fe was (1.40±0.46) g·L-1.The preparation exhibited apparent superparamagnetism and the saturation magnetization was 7.75 A·m2·kg-1.The entrapment efficiencies of quercetin and resveratrol in QR-MSLN were 99.10% and 80.83%,respectively.QR-MSLN had a significantly higher effect of tumor inhibition than SLN(containing quercetin and resveratrol) and free drug(PConclusion:QR-MSLN has uniform particle size and good magnetic response,and shows remarkable antitumor effect on H22 bearing mice.
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Objective: To explore the effect and mechanism of genistein on oleic acid-induced lipid accumulation in HepG2 cells. Method: Lipid accumulation model in HepG2 cells was induced by different concentrations of oleic acid for 24 h, and 12.5, 25, 50 μmol·L-1genistein and oleic acid acted on cells for 24 h. Cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. Double staining with Nile red and DAPI was used to observe the intracellular lipid droplets. Intracellular triglyceride (TG) content was determined by kit. The protein expression levels of triglyceride lipase(ATGL),hormone-sensitive fatty acid(HSL),phosphorylation HSL(p-HSL),silent information regulator 1(STRT1),peroxisome proliferator-activated receptor α(PPARα),carnitine palmityl transferase 1(CPT-1) in oleic acid-induced HepG2 cells were detected by Western blot. Result: 0.5 mmol·L-1 oleic acid and 12.5, 25, 50 μmol·L-1 genistein had no significant effect on cell viability after treated cells for 24 h. Compared with normal group, the TG content and lipid droplets in oleic acid-induced HepG2 cells was significantly increased (PPPPα, and CPT-1 compared with model group (PPConclusion: Genistein can significantly improve the lipid accumulation in oleic acid-induced HepG2 cells, and its mechanism may be related to up-regulating the protein expression levels of ATGL, p-HSL/HSL, SIRT1, PPARα, CPT-1, and thus promoting lipid hydrolysis and oxidative metabolism.
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@# Objective： ：To study the regulatory effect of mogrol (MO) on lipid metabolism of hepatic cancer cells and its molecular mechanism. Methods: Oleic acid (OA) was used to induce fat accumulation in hepatocellular carcinoma HepG2 cells and to establish a steatosis cell model. CCK-8 method was used to detect the cytotoxicity of MO to HepG2 cells, and an experimental working concentration without obvious cytotoxicity of MO was chosen. After being treated with different concentrations of MO, lipid accumulation in the cells was observed by oil red O staining, and the contents of triglyceride (TG) and cholesterol (TC) in the cells were measured. Key genes involving in lipid metabolism were screened out by high-throughput transcriptome sequencing qPCR was used to detect the mRNA expressions of ,SREBP-1c and FASN, while Western Blot was used to detect the protein expressions of p-AMPKα, SREBP-1c and FASN in cells of model group and treatment group. Results: After OA induction, a large amount of lipids accumulated in HepG2 cells, the contents of TG and TC increased significantly. Three key genes (SREBP-1c, FASN and p-AMPK α) involving in lipid metabolism of hepatic cancer cells were screened out. After OA induction, the mRNA expressions of SREBP-1c and FASN increased, the protein expression of p-AMPK α decreased while the protein expressions of SREBP-1c, FASN and other proteins increased significantly. After intervention with working concentration of MO, intracellular lipid accumulation, contents of TG and TC, mRNA expressions of SREBP-1c, FASN and protein expressions of SREBP-1c, FASN decreased significantly, while the expression of p-AMPKα increased. Conclusion: Mogrol can inhibit the synthesis of fatty acids by activating the expression level of AMPK signaling pathway related factors SREBP-1c and FASN, so as to play the role of regulating lipid metabolism.
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OBJECTIVE： To study the improvement effect and mechanism of methylated urolithin A on oleic acid-induced lipid accumulation in human liver cancer Huh-7 cells. METHODS： Oleic acid was adopted to induce lipid accumulation model cells. Huh-7 cells were divided into control group （culture medium）， model group （1 mmol/L oleic acid）， low-dose group （1 mmol/L oleic acid+10 μmol/L methylated urolithin A） and high-dose group （1 mmol/L oleic acid+20 μmol/L methylated urolithin A）. Oil red O staining was used to observe lipid accumulation in cells. Triglyceride（TG） enzyme assay was applied to determine the TG content in cells. PCR was employed to detect the mRNA expression of FASN， SREBP-1， PPAR-α and PPAR-γ in cells. Western blotting was used to determine the protein expression of FASN in cells. RESULTS： After induced by oleic acid， a large amount of lipid droplet accumulated around the cells； the intracellular lipid and TG content， mRNA expression levels of FASN， SREBP-1 and PPAR-γ， protein expression levels of FASN were increased significantly， while mRNA expression level of PPAR-α was decreased significantly （P＜0.01）. After intervened with methylated urolithin A， lipid droplet around the cells decreased significantly； the contents of lipid and TG in cells were decreased significantly， while the mRNA expression levels of FASN， SREBP-1 and PPARγ and protein expression level of FASN were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Methylated urolithin A can improve oleic acid-induced lipid accumulation in Huh-7 cells， the mechanism of which may be associated with inhibiting fat synthesis， promoting lipid metabolism and down-regulating the expression of metabolism-related factors as FASN， SREBP-1 and PPAR-γ.
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Yuhua91 is a new peanut variety with high oleic acid content bred by Qingdao Agricultural University. The crossing was conducted with Luhua11 as female parent and with Kainong1715, an F435-type variety with high oleic acid content as male parent. The real F1 hybrids were screened by sequencing on PCR amplification products, and those homozygotes with bb genotype in F2 populations were screened by the same sequencing method as above. The content of oleic and linoleic acid was measured on the kernels harvested from F2 single plants by near infrared ray method, and those kernels whose content of oleic was above 80%, oleic and linoleic acid ratio was above 10.0 were obtained and planted into a row, with pedigree method for subsequent selection breeding. Yuhua91 has some characters of small pod, light and obvious pod texture, 148.06 g per 100 pods, 63.31 g per 100 kernels, 75.15% shelling percentage, long elliptic seed kernel, pink seed coat, without crack, white endotesta. Its content of protein, oil, oleic acid, linoleic acid and palmitic acid was 26.57%, 52.72%, 80.40%, 2.50% and 5.57% respectively. Yuhua91 has other characters of strong seedlings, compact pod areas, and moderate resistance to leaf spot disease and bacterial wilt. Average pod yield is 215.79 kg per Mu, 15.27% higher than the control variety Huayu20. Average seed kernels yield is 157.33 kg per Mu, 21.64% higher than the control variety Huayu20. Yuhua 91 has been registered on department of agriculture in 2018, and the registration No. is GPD peanut (2018) 370210, fit for growing in Shandong Province.
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Arachis , Oleic Acid , Plant Breeding , SeedsABSTRACT
Objective@#To establish a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the detection of serum oleic acid (OA), and preliminarily evaluate the role of OA in insulin resistance (IR) of type 2 diabetes (T2DM). @*Methods@#OA-［ 13 C 5 ］ was used as isotope-labeled internal standard, and the ion pairs of OA and OA-［ 13 C 5 ］ were 281.3/281.3 and 286.3/286.3, respectively. The ultrapure water was used as mobile phase A and methanol: acetonitrile (1∶1, v/v) as mobile phase B in a ZORBAX SB-Aq C18 reversed phase column. Meanwhile, the gradient elution system with a flow rate of 0.3 mL/min was used. According to the CLSI guidelines (EP15-A3), the reliability of the established method was evaluated by detecting the performance indicators such as precision, trueness, linear range, stability and carrying contamination rate. Serum OA levels were detected by the established HPLC-MS/MS method in 109 patients with clinically diagnosed T2DM and 100 healthy controls. The insulin resistance index (HOMA-IR) was calculated to evaluate IR, and the relationship between OA and IR was further analyzed. @*Results@#The established HPLC-MS/MS method for the detection of serum OA had good specificity and linearity in the range of 10-1 000 μmol/L (y=0.007 55x+0.004 83,r=0.997 7), and the low limit of quantification (LLOQ) was 10 μmol/L. It also had good precision, and the within-run coefficient of variation (CV) and total CV were not more than 1.62% and 1.73%, respectively, indicating that the method was suitable for the detection of serum OA. The serum OA levels in T2DM patients ［(425.58 ± 220.17) μmol/L］ were significantly higher than that in the healthy controls ［(113.20±58.00) μmol/L］, and serum OA levels were significantly correlated with HOMA-IR in T2DM patients and healthy controls. The area under the receiver operating characteristic (ROC) curve (AUC) of OA for the diagnosis of IR was 0.689. When the cut-off value identified by Youden index was 235.8 μmol/L, the sensitivity and specificity were 70.4% and 63%, respectively. When OA combined with fasting blood glucose (FBG) to diagnose IR, the AUC increased to 0.806, which was significantly higher than that of OA (P＜0.05). @*Conclusion@#A scientific and efficient HPLC-MS/MS method for the quantitative detection of serum OA is established successfully, which provides a reliable method for the dynamic monitoring of the changes of OA levels in the patients with metabolic diseases.
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OBJECTIVE: To screen pressure sensitive adhesive system and penetration enhancer of rutaecarpine transdermal patch. METHODS: The patch was prepared by solvent evaporation method. The ratio between pressure sensitive adhesive (Eudragit E100), crosslinking agent (succinic acid) and plasticizer (dibutyl sebacate) of the rutaecarpine patch were screened through Box-Behnken design by using adhesion (stick power, shear strength, peel force) as the index. The ratio between pressure sensitive adhesives and chemical enhancers (azone and oleic acid) were screened by adhesion and permeation experiments in vitro using custom design, which were carried out by using improved Franz diffusion cells through excised mice skin. RESULTS: The optimized formulation of rutaecarpine transdermal patch consisted of 83% pressure sensitive adhesives (63.5% Eudragit E100, 5.5% succinic acid, 14% dibutyl sebacate), 10% azone, 6.4% oleic acid and 0.6% rutaecarpine. The stick power, shear strength, and peel force of the patch were 15 steel balls, (10.97 ± 0.32) h and (0.16 ± 0.02) kN•M-1, respectively. The cumulative permeation amount and permeation rate of the patch were (29.71 ± 1.19) μg•cm-2 and (1.36 ± 0.10) μg•cm -2•h -1, respectively. CONCLUSION: The optimized rutaecarpine patch show increased permeation and appropriate adhesion. This study provides the basis for future research.
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Objective@#To explore the effects of oleic acid and palmitic acid on lipid deposition and mTOR/S6K1/SREBP-1c pathways in HepG2 cells.@*Methods@#The model of steatosis was established with induction of oleic acid and palmitic acid and was intervened by rapamycin. The changes in lipid droplets were observed after staining the cells with oil Red O. Intracellular triglyceride (TG) contents in cells were measured by TG kit. mTOR, S6K1, and SREBP-1c mRNA expression levels were detected using QRT-PCR. Western blot was used to determine protein expression levels of mTOR, S6K1 and SREBP-1c.@*Results@#Both fatty acids increased lipid droplets in HepG2 cells. Fatty degeneration with elevated TG occurred with significant changes in oleic acid group lipids. Rapamycin alleviated lipid deposition caused by oleic acid and palmitic acid and inhibited their induction of increased expression of mTOR, S6K1, and SREBP-1c. QRT-PCR and Western blot results showed that mRNA and protein expressions of mTOR, S6K1, and SREBP-1c in oleic acid and palmitic acid group were significantly higher than the control group (P < 0.05). The increase was more pronounced in the palmitic acid group (P < 0.05); however, after rapamycin intervention, the expression of mRNA and protein in the three groups were significantly lower (P < 0.05), and the change in palmitic acid group was more pronounced (P < 0.05).@*Conclusion@#Oleic acid and palmitic acid can induce lipid deposition in HepG2 cells and increase expression of every component of mTOR/S6K1/SREBP-1c pathway; however, Oleic acid-induced lipid deposition is more pronounced, and the mTOR, S6K1, and SREBP-1c pathway change is more obvious in palmitic acid. Rapamycin has high potent inhibitory effect on palmitic acid-induced lipid deposition. These results specify that lipid synthesis involved in the mTOR/S6K1/SREBP-1c pathways are mainly associated to palmitic acid in HepG2 cells, whereas other signaling pathway may mediate oleic acid-induced lipid synthesis.
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Objective@#To establish a new model of hepatic steatosis cells by optimizing the original ethanol or high fat, the present study proposed an in vitro hepatocyte steatosis model for the study of fatty liver.@*Methods@#Oil red O staining was used to observe the effects of fetal bovine serum, oleic acid and ethanol on lipid accumulation in human liver cell line L02 in a concentration- and time-dependent manner. RT-PCR was used to detect the mRNA expression levels of PPAR-γ and AP-2, and the suitable conditions for the establishment of hepatocyte steatosis model were screened out. A t-test was used for comparison between the two groups, and one-way Analysis of Variance (ANOVA) was used in more than three groups.@*Results@#Oil red O staining showed the number of reddish-orange lipid droplets in L02 cells gradually increased with the increase of fetal bovine serum, oleic acid and ethanol in a concentration - and time-dependent manner. Compared with 0.00% oleic acid and 2% ethanol, the count value of red particle was 100.00% ± 17.63% at the beginning and after 24 h, 0.003% oleic acid and 2% ethanol jointly acted in L02 cells. After incubation for 48 hours with 2% ethanol and serum-free DMEM medium, the accumulation of lipid droplets was the highest with a count value of 802.38%+71.06%(t = 42.36, P < 0.001). RT-PCR analysis showed the lipid accumulation induced by this method was positively correlated with the mRNA expression of PPAR-γ and AP-2.@*Conclusion@#L02 cells were successfully exposed to high fat and ethanol, and the hepatocyte steatosis model was established and optimized, suggesting that the occurrence of hepatic cell steatosis was related to the up-regulation of PPAR-γ and AP-2.
ABSTRACT
Objective: To study the dose-effect relationship of Yuning ointment and its decomposed recipes in the treatment of oleic acid induced acne in mice. Methods: Oleic acid was administrated to the back (2 cm ×2 cm) of the mice (once a day) for 21 days to induce acne. At d22, the gradient dosage of Anemone flaccida crude drug (1. 06-1 060. 23 mg?kg-1?d-1,k=3. 16), Yuning oint-ment without Anemone flaccida crude drug (4. 73-1 767. 75 mg?kg-1?d-1, k=3. 16) and Yuning ointment (2. 84-2 827. 28 mg?kg-1?d-1, k=3. 16) was respectively administrated to the back of mice for 14 days. The pathological changes of skin were observed by hematoxylin-eosin (HE) staining. The diameter of sebaceous glands and the ratio of follicular keratinization area were morphomet-rically analyzed. The serum levels of IL-1, IL-6 and TNF-α were detected by ELISA assay. The median effective dosages (ED50) of A-nemone flaccida in the three prescriptions were regressed by Prism 5. 01 software to determine the prescription dose-effect. Results: All the therapy groups were with significantly relieved pathological changes of sebaceous glands hypertrophy and follicular keratinization, and decreased serum levels of IL-1, IL-6 and TNF-α in a dose-dependent manner. The dose-response curves showed an "S" shape. A-mong the three therapy groups, the effect of Yuning ointment was the best. The ED50of Yuning ointment regressed by Anemone flaccida dose was 0. 28-fold for improving sebaceous glands hypertrophy, 0. 14-fold for inhibiting follicular keratinization, and 0. 15-, 0. 49-and 0. 24-fold for decreasing serum levels of IL-1, IL-6 and TNF-α. . Regressed by Yuning ointment without Anemone flaccida, the ED50of Yuning ointment was lower than Yuning ointment without Anemone flaccid in terms of improving pathological changes and inhibiting the secretion of cytokines. Conclusion: Yuning ointment can prevent and treat acne through regulating immune function. And the prescrip-tion compatibility can enhance the effects of Anemone flaccida.