Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment     

DAI Pei  GAO Fen  GAO Hong-wei  WANG Yuan  FENG Gao-jie  ZHANG Qin-feng  BAI Rui  QIN Wei-wei  LI Hong  SONG Xiao-su 《园艺学报》2019,35(2):212-217

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.  相似文献   

Effect of 4-chloro-2-methylphenoxyacetic acid and 2,4-dimethylphenol on human erythrocytes     

Bo?ena Bukowska  Katarzyna GoszczyńskaWirgiliusz Duda 《Pesticide biochemistry and physiology》2003,77(3):92-98

The effects of exposure of human erythrocytes to different concentrations of 4-chloro-2-methylphenoxyacetic acid (MCPA) and its metabolite—2,4-dimethylphenol (2,4-DMP) were studied. The investigations concerned mainly the content of glutathione (GSH and GSSG), glutathione peroxidase (GSH-Px), glutathione transferase (GST), and the level of adenine energy charge (AEC). Reactive oxygen species (ROS) such as hydroxyl radical, superoxide anion, hydrogen peroxide, and nitric oxide are produced during normal processes in the cell. Under normal conditions, antioxidant systems of the cell minimize damage caused by ROS. When ROS generation increases to an extent that it overcomes the cellular antioxidant systems, the result is oxidative stress. We observed that MCPA and 2,4-DMP decreased the level of GSH in erythrocytes in comparison with control. MCPA did not affect glutathione peroxidase and glutathione transferase activity, while 2,4-DMP increased their activity. 2,4-DMP decreased the level of ATP and increased the content of ADP and AMP, leading to the fall of the level of AEC. MCPA and 2,4-DMP transform hemoglobin into methemoglobin, thus preventing oxygen transport. Comparison of the toxicity of MCPA and 2,4-DMP revealed that the most prominent changes occurred in human erythrocytes incubated with 2,4-DMP.  相似文献   

Guanidinoacetic acid supplementation in broiler chickens fed on corn-soybean diets affects performance in the finisher period and energy metabolites in breast muscle independent of diet nutrient density     

M. Majdeddin  H. Kermanshahi  S. De Smet  J. Michiels 《British poultry science》2018,59(4):443-451

(1) Guanidinoacetic acid (GAA) is the single immediate endogenous precursor of creatine (Cr). It was hypothesised that dietary GAA would have different effects on performance and energy metabolites in breast muscle depending on the nutrient density (ND) of corn-soybean-based diets.

(2) A total of 540 one-day-old male Ross 308 broilers were allocated to 9 dietary treatments with 6 replicates (10 birds each) in a 3 × 3 factorial arrangement with three levels of ND (low, 2800; medium, 2950 and high, 3100 kcal metabolizable energy (ME)/kg; and with the other nutrients being constant relative to ME) and supplemented with three levels of GAA (0, 0.6 and 1.2 g/kg) in a 42-d feeding trial.

(3) In the starter and grower periods, increasing levels of ND improved body weight (BW), average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR), with the exception of ADFI in the starter period. GAA supplementation did not affect performance characteristics. All performance indicators responded markedly to increasing ND in the finisher period, whereas the highest GAA level reduced ADFI compared to the unsupplemented control (156 vs. 162 g/d) and concomitantly FCR (1.81 vs. 1.93). No interactive effects were noted for any performance trait. The high ND diet resulted in more breast meat yield on d42, associated with higher fat content and darker colour compared to the other ND levels. The GAA supplementation did not affect carcass and breast traits. At the end of the experiment, Cr was elevated when feeding GAA at 1.2 g/kg (5455 vs. 4338 mg/kg fresh muscle).

(4) To conclude, ND had a substantial effect on performance and carcass traits, whereas any effect of GAA was limited to FCR in the finisher period and independent of diet ND level.  相似文献   

Lipid-rich bovine serum albumin improves the viability and hatching ability of porcine blastocysts produced in vitro     

Chie SUZUKI  Yosuke SAKAGUCHI  Hiroyoshi HOSHI  Koji YOSHIOKA 《The Journal of reproduction and development》2016,62(1):79-86

The effects of lipid-rich bovine serum albumin (LR-BSA) on the development of porcine blastocysts produced in vitro were examined. Addition of 0.5 to 5 mg/ml LR-BSA to porcine blastocyst medium (PBM) from Day 5 (Day 0 = in vitro fertilization) significantly increased the hatching rates of blastocysts on Day 7 and the total cell numbers in Day-7 blastocysts. When Day-5 blastocysts were cultured with PBM alone, PBM containing LR-BSA, recombinant human serum albumin or fatty acid-free BSA, addition of LR-BSA significantly enhanced hatching rates and the cell number in blastocysts that survived compared with other treatments. The diameter, ATP content and numbers of both inner cell mass and total cells in Day-6 and Day-7 blastocysts cultured with PBM containing LR-BSA were significantly higher than in blastocysts cultured with PBM alone, whereas LR-BSA had no effect on mitochondrial membrane potential. The mRNA levels of enzymes involved in fatty acid metabolism and β-oxidation (ACSL1, ACSL3, CPT1, CPT2 and KAT) in Day-7 blastocysts were significantly upregulated by the addition of LR-BSA. The results indicated that LR-BSA enhanced hatching ability and quality of porcine blastocysts produced in vitro, as determined by ATP content, blastocyst diameter and expression levels of the specific genes, suggesting that the stimulatory effects of LR-BSA arise from lipids bound to albumin.  相似文献   

Purinergic control of the quail rectum: modulation of adenosine 5'-triphosphate-mediated contraction with acetylcholine     

Shiina T  El-Mahmoudy A  Khalifa M  Draid M  Shimizu Y  Takewaki T 《Research in veterinary science》2007,82(2):246-251

Electrical field stimulation (EFS) induces frequency-dependent contractions of the longitudinal muscle of isolated quail rectum which were sensitive to tetrodotoxin. The aim of the present study was to investigate whether purinergic neurons are implicated in the response to nerve stimulation. The shape of the EFS-induced contractile response was different depending on stimulus frequency; low frequencies (0.5-2 Hz) induced fast monophasic contractions with a small subsequent relaxation; whereas higher frequencies (5-50 Hz) induced biphasic contractile response that comprised fast initial component (as in case of low frequency) and a slow delayed contractile component in addition to the relaxation that follows the fast contractile component. Prior application of atropine (10 microM) completely abolished the slow delayed component but significantly enhanced the fast initial contractile component. Physostigmine (1-10 microM) significantly enhanced the slow delayed component with an inhibitory effect on the initial fast component. The nonspecific purinergic receptor antagonist, suramin (100-500 microM) significantly inhibited the fast initial contractile component with no significant effect on the slow delayed one. Complete blockade of the fast component was achieved by prior application of a combination consisted of suramin (50 microM) and pyridoxicalphosphate-6-azophenyl 2',4'-disulphonic acid tetrasodium (PPADS; 10 microM). Exogenous applications of adenosine 5'-triphosphate and acetylcholine (10 microM each), produced contractile responses that mimicked those induced by EFS. These data suggest that ATP is the main noncholinergic excitatory transmitter controlling the contractile activity of the quail rectum; and that its action could be modulated by acetylcholine.  相似文献   

红莲型CMS水稻线粒体K+ATP通道特性的初步研究     

夏春皎  王卓  徐鑫  马亢  刘学群  王春台 《华中农业大学学报》2006,25(3):219-222

以水稻红莲型细胞质雄性不育(HL-CMS)不育系粤泰A(YTA)和保持系粤泰B(YTB)黄化苗线粒体为材料,研究了YTA和YTB离体线粒体KA TP通道对其诱导调节剂KCl、ATP、ADP和GTP等的响应特性。结果表明,KA TP通道诱导剂KCl,能明显诱导YTA和YTB线粒体膨胀,但YTA的膨胀程度较YTB的明显;KA TP通道的内源性抑制剂ATP对YTA和YTB线粒体膨胀起显著抑制作用;KA TP通道的内源性激动剂ADP和GTP引起的不育系YTA离体线粒体短暂收缩及之后的再膨胀程度均较YTB的明显;此外,比较YTA和YTB离体线粒体在Rh123一起孵育10 min后,KCl和解偶联剂FCCP处理引起的离体线粒体膜电位(Δψm)下降过程,发现前者的线粒体Δψm较后者的易于过早崩解。对水稻HL-CMS不育系YTA和可育的保持系YTB线粒体KA TP通道特性的比较研究表明,水稻HL-CMS不育系YTA线粒体KA TP通道对其诱导调节剂KCl、ATP、ADP和GTP等的响应较YTB的更敏感。  相似文献   

猪冷冻精液的研究   总被引：1，自引：0，他引：1  

曾维斌  王志刚  刘丑生  张兆旺  赵俊金 《中国畜牧兽医》2005,32(5):36-38

本试验以甘油、乙二醇为抗冻剂，并添加ATP或安息香酸咖啡因，以解冻后精子活力、顶体完整率、顶体膨胀率、顶体破损率、尾部畸形率和在37℃下的存活时间几个重要评定指标为依据，筛选了几种猪颗粒冻精的冷冻稀释液配方，并优化了其冷冻程序。结果表明，与对照组相比，在冷冻液中添加0．1mg／ml的ATP或2mg／ml的安息香酸咖啡因都能显著提高解冻后精子活力(分别提高0．13和0．18)，降低顶体膨胀率(两者都将近降低了11％)，并能延长其存活时间(分别延长10．5h和11．8h)；甘油和乙二醇两种抗冻剂配合使用，将显著提高冻后精子的活力与存活时间，其较优混合量为甘油3％，乙二醇0．5％。  相似文献   

不同来源ATP表征冷鲜羊肉新鲜度     

苏媛媛  张德权  古明辉  张春娟  李少博  郑晓春  陈丽 《中国农业科学》2022,55(19):3841-3853

【目的】通过系统研究冷鲜羊肉不同来源的三磷酸腺苷（adenosine triphosphate,ATP。包括肉中ATP、微生物ATP、肉表面ATP）在贮藏期间的变化规律,筛选能够表征冷鲜羊肉新鲜度变化的ATP指标,构建菌落总数和挥发性盐基氮预测模型,探究冷鲜羊肉新鲜度的预测新方法。【方法】以小尾寒羊背最长肌为试验材料,在空气密封包装0℃条件下分别贮藏0、1、3、5、7、9、11、13、15、17和21 d,分析冷鲜羊肉贮藏期间新鲜度指标（pH、色泽、挥发性盐基氮、菌落总数）与3种来源ATP（肉中ATP、微生物ATP、肉表面ATP）的变化,利用数据统计评价不同来源ATP的变化规律,并构建新鲜度指标的预测模型。【结果】冷鲜羊肉贮藏期间新鲜度指标菌落总数、挥发性盐基氮均呈现上升趋势,并均在贮藏17 d时超过国家标准限值;肉中ATP呈现不断下降趋势,微生物ATP与肉表面ATP均呈现上升趋势,与新鲜度指标变化趋势保持一致;冷鲜羊肉贮藏期间,肉中ATP、微生物ATP、肉表面ATP含量与菌落总数、挥发性盐基氮的相关系数（R）分别为-0.399、0.910、0.943和-0.357、0.725、0.907。肉表面ATP预测冷鲜羊肉菌落总数的最优模型为Boltzmann拟合模型,其公式为TVC（lg cfu/g）=7.649-4.069/(1+exp(x-5.807)/0.632)（R ²=0.903,P<0.001）,肉表面ATP预测冷鲜羊肉挥发性盐基氮的最优模型为Expedc1拟合模型,其公式为TVB-N（mg/100 g）=2.493*exp(x/3.745)+3.057（R ²=0.888,P<0.001）。【结论】本研究明确了冷鲜羊肉表面ATP与菌落总数、挥发性盐基氮存在显著正相关性,确证了肉表面ATP可以作为冷鲜羊肉新鲜度表征指标;并构建了菌落总数和挥发性盐基氮最优预测模型,为冷鲜羊肉新鲜度快速检测提供了新的思路。  相似文献   

A study of goldfish oocyte meiosisin vitro: effects of 2,4-dinitrophenol and adenosine-5-triphosphate     

Hamid R. Habibi  Charles A. Lessman 《Fish physiology and biochemistry》1986,1(4):197-205

Germinal vesicle migration (GVM) and/or dissolution (GVD) were measured in goldfish oocytes, treated with 17α, 20β dihydroxyprogesterone (DHP) and other compounds considered to effect the cytoskeleton and oxidative phosphorylation,in vitro. Administration of DHP reinitiated meiotic maturation, increasing GVM and GVD in goldfish oocytes. Addition of 2,4-dinitrophenol (DNP) to the incubation medium significantly inhibited DHP-induced GVM and GVD. The DNP effect was found to be partially reversible after 24 h and could be reversed fully after a further delay of approximately 24h. Treatment of goldfish oocytes with demecolcine (DE; a colchicine derivative also known as colcemid) induces GVM to the micropyle without effecting GVD; while Cytochalasin-B which inhibits microfilament polymerization impairs both GVM and GVD. Administration of DNP, significantly inhibited DE-induced GVM, suggesting that GVM as well as GVD are dependent upon the process of oxidative phosphorylation. Addition of adenosine-5′ -triphosphate (ATP) at low concentrations (0.01–0.1 mM) did not effect DHP-induced or DNP-inhibited GVD in goldfish oocytes. The present results are consistent with the idea that migration of the oocyte nucleus during meiosis reinitiation has an energy requirement and involves participation by the cytoskeleton.  相似文献   

Are Primary Cultures of Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) Ventricular Cardiomyocytes Metabolically Viable?     

J.P?Goldstein  A.?Scott  S.?CurrieEmail author 《Fish physiology and biochemistry》2004,30(2):109-117

The objectives of this study were to establish a protocol for the isolation of metabolically viable ventricular cardiomyocytes from the rainbow trout and to determine which measures may best reflect viability 24 h after isolation. Cardiomyocytes were isolated by enzymatic digestion and maintained in cell suspension. Viability was assessed using Trypan blue dye exclusion, ATP content, oxygen consumption and lactate dehydrogenase (LDH) leakage into the medium. Viability, assessed by these measures did not significantly change over the time period of this study. ATP content did correlate significantly with oxygen consumption but not with Trypan blue exclusion. We conclude that primary cultured cardiomyocytes remain metabolically viable for at least 24 h after isolation. Also, it appears that ATP content and oxygen consumption most adequately reflect metabolic cell viability. To be confident with a culture, however, a combination of viability measures is necessary when isolating cardiomyocytes from fish.  相似文献