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1.
旨在研究AMPK激活剂二甲双胍（metformin，Met）和阿卡地新（acadesine，AICAR）对绵羊精液冷冻保存效果的影响。本研究首先在冷冻基础稀释液中分别添加不同浓度（0、100、200、300、400、500 μmol·L-1）的Met和AICAR，冷冻解冻后根据精子活力、运动性能和结构完整性指标筛选出最佳的添加浓度（400 μmol·L-1 Met、200 μmol·L-1 AICAR）；然后分别使用不同的冷冻稀释液（对照组：稀释液；Met组：含400 μmol·L-1 Met的稀释液；AICAR组：含200 μmol·L-1 AICAR的稀释液）冷冻精液，解冻后检测精子中AMPK蛋白表达、顶体酶活性、代谢指标、线粒体功能以及抗氧化酶活性。结果表明，稀释液中添加400 μmol·L-1 Met和200 μmol·L-1AICAR均可显著提高解冻后精子活力、运动性能及精子结构完整性（P<0.05），其中400 μmol·L-1 Met组精子总活力达43.20%，顶体完整率为91%，质膜完整率为46%。与对照组相比，Met组和AICAR组解冻后精子中AMPK磷酸化水平显著升高（P<0.05）；顶体酶活性显著提高（P<0.05）；丙酮酸水平显著下降（P<0.05），乳酸脱氢酶活性、乳酸以及ATP含量均显著升高（P<0.05）；与对照组相比，Met和AICAR组稀释液更有利于维持线粒体膜电位（P<0.05），提高ATP酶（P<0.05）以及抗氧化酶的活性（P<0.05）。添加适当浓度的AMPK激活剂可以提高绵羊精液冷冻保存的效果。 相似文献
2.
本试验旨在研究植物提取物白藜芦醇(RES)对牛皮下脂肪细胞凋亡率以及去乙酰化酶1(SIRT1)/腺苷一磷酸激活的蛋白激酶(AMPK)信号通路关键因子的mRNA和蛋白质表达量的影响。选取18月龄鲁西黄牛的皮下前体脂肪细胞,在细胞分化的第0天,更换为RES浓度分别为0(对照)、100、200和400μmol/L的培养液处理48 h,每组设3个重复。应用Hoechst33342染色检测细胞凋亡的形态学变化,流式细胞分析仪检测细胞凋亡率,荧光定量PCR(q PCR)和免疫印迹试验(Western-blot)分别检测SIRT1/AMPK信号通路上的关键因子SIRT1、A MPKα、叉头转录因子1(Fox O1)、B细胞淋巴瘤/白血病-2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、促凋亡蛋白Bcl-2相关X蛋白(Bax)的mRNA和蛋白质表达量,并利用油红O染色鉴定脂肪细胞。结果表明,与对照组相比:不同浓度RES处理后的牛皮下脂肪细胞凋亡率均极显著增高(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3、Bax的mRNA表达量均显著或极显著提高(P0.05或P0.01),Bcl-2的mRNA表达量极显著降低(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3的蛋白质表达量均显著或极显著提高(P0.05或P0.01);200和400μmol/L RES处理后,Fox O1的mRNA和蛋白质表达量显著或极显著提高(P0.05或P0.01),Bax的蛋白质表达量极显著提高(P0.01),Bcl-2的蛋白质表达量极显著降低(P0.01)。100μmol/L RES处理后时,Fox O1的mRNA和蛋白质表达量以及Bcl-2、Bax的蛋白质表达量均与对照组差异不显著(P0.05)。由此可见,RES通过激活SIRT1/AM PK信号通路,同时激活通路下游的Fox O1,促进了牛皮下脂肪细胞的凋亡,为通过营养调控技术降低牛皮下脂肪沉积提供了一定的理论基础。 相似文献
3.
旨在探究产蛋各期番鸭肝腺苷酸活化蛋白激酶（AMPK）信号通路基因表达和肝脂肪酸组成,为番鸭肝脂质代谢应答产蛋提供机理研究。本研究选取开产前22周龄、产蛋初期30周龄、产蛋中期40周龄和产蛋末期60周龄母番鸭各15羽,全自动生化仪测定血脂水平,苏木精-伊红（HE）染色和油红O染色观察肝组织学结构,实时荧光定量PCR （qRT-PCR）检测肝AMPK通路基因表达,气相色谱-质谱联用法（GC-MS）检测产蛋各期肝脂肪酸组成。结果表明,总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、甘油三酯（TG）和极低密度脂蛋白胆固醇（VLDL-C）水平在40周龄显著高于22、30和60周龄（P<0.05）;高密度脂蛋白胆固醇（HDL-C）水平在30和40周龄显著低于22和60周龄（P<0.05）。肝HE和油红O染色切片显示,肝在22周龄呈实质状,至产蛋40和60周龄,肝脂滴沉积明显（P<0.05）。AMPKα1在22、30、40和60周龄呈本底低水平表达,并显著低于产蛋各期肉碱脂酰转移酶1（CPT1）和脂肪酸合成酶（FAS）的表达量（P<0.05）,FAS在22、40和60周龄均呈高水平表达（P<0.05）;羟甲基戊二酰辅酶A还原酶（HMGR）、肝细胞核因子4α（HNF4α）、乙酰辅酶A羧化酶1（ACC1）和固醇调控元件结合蛋白-1（SREBP1c）在40周龄表达量显著高于22和30周龄（P<0.05）。肝中饱和脂肪酸（SFA）、单不饱和脂肪酸（MUFA）和多不饱和脂肪酸（PUFA）主要由C16∶0、C18∶0、C18∶1和C20∶2 n-6构成,分别占总脂肪酸含量的32%、16%、30%和9%。C14∶0和C16∶0含量在40周龄显著高于22周龄（P<0.05）;C24∶0、C20∶2 n-6和PUFA含量在60周龄显著高于22和40周龄（P<0.05）。综上,番鸭产蛋期肝通过上调FAS等脂质合成基因表达,合成大量长链脂肪酸,沉积于肝,并增加血脂水平。 相似文献
4.
《The Journal of Applied Poultry Research》2016,25(3):370-378
The objective of this experiment was to evaluate the effects of dietary glutamine and glucose on pectoralis major meat quality in broilers under acute heat stress. Three hundred 35-day-old male broilers (Arbor Acres) were randomly divided into 5 groups. Broilers in the positive control group were kept in a thermoneutral environment (23 ± 1°C, RH 45%–55%) and fed a basic diet, whereas the experimental (2 × 2) group was exposed to acute heat stress (34 ± 1°C, RH 65%–75%) for 12 h and fed the basic diet that contained glutamine (0 and 10 g/kg) and glucose (0 and 10 g/kg). Compared with the positive control group, acute heat stress without glutamine and glucose supplementation increased (P < 0.05) drip loss, L* values, and AMPK concentrations, but it decreased (P < 0.05) water-holding capacity, moisture, pH, a* values, b* values, and glutamine concentrations in pectoralis major meat of chickens. However, dietary glutamine (10 g/kg) increased (P < 0.05) water-holding capacity, moisture, pH, a* values, b* values, and glutamine concentrations, while it decreased (P < 0.05) drip loss, L* values, and AMPK concentrations in heat-stressed chicken pectoralis major meat. Dietary glucose (10 g/kg) increased (P < 0.05) water-holding capacity, moisture, pH, and a* and b* values, while it decreased (P < 0.05) L* values and AMPK concentrations of pectoralis major meat in chickens exposed to acute high temperature. There were significant interactions between glutamine and glucose in the water-holding capacity, pH values, AMPK, and glutamine levels of pectoralis major meat in the heat-stressed chickens. The above results suggest that the addition of glutamine and glucose in the diet is necessary for broilers during acute heat stress condition. 相似文献
5.
本试验旨在考察洛伐他汀的降胆固醇作用是否由一磷酸腺苷激活的蛋白激酶(AMPK)所介导.选用健康的产蛋率、体重相近的47周龄新罗曼蛋鸡20只,分为2个处理,每个处理5个重复,每个重复2只鸡,单笼饲养,分别饲喂基础日粮(对照组)和试验日粮(基础日粮中添加0.06%洛伐他汀),正式试验期为3周.结果表明:1)添加0.06%洛伐他汀不影响产蛋鸡生产性能,并可以降低鸡蛋胆固醇含量.与对照组相比,按每克蛋黄胆固醇含量(mg/g蛋黄)计,第1、2、3周分别降低11.56%、10.88%和12.75%(P＜0.05);按全蛋总胆固醇(mg/个蛋黄)计,第1、2、3周分别降低16.01%、17.95%和19.79%(P＜0.05);2)添加0.06%洛伐他汀,使产蛋鸡肝脏AMPK活性提高5.69%(P＞0.05),β-羟基-β-甲基-戊二酸单酰辅酶A还原酶(HMGR)、乙酰辅酶A羧化酶(ACC)活性分别降低21.89%(P＜0.05)和9.72%(P＞0.05);3)细胞培养液中加入0.3 mmol/L洛伐他汀,产蛋鸡肝细胞AMPK活性提高5.02%(P＞0.05),同时加入0.3 mmol/L洛伐他汀和0.5 mmol/L Br8-AMP(AMPK的抑制剂)时,AMPK活性较洛伐他汀组降低4.57%(P＞0.05).由此可知,洛伐他汀的降胆固醇作用可能部分由AMPK介导. 相似文献
6.
【目的】探究咖啡酸苯乙酯（caffeic acid phenethyl ester,CAPE）通过AMPK/FOXO3a信号通路缓解氧化应激对猪精液的影响及其分子机制。【方法】选取8头成年（1—2岁）、健康状况良好长白种公猪进行鲜精采集,将质检合格的样品混池待用。试验设计如下：首先,在常温基础稀释液中分别添加0、0.02、0.04、0.06、0.08和0.10 g·L-1浓度的CAPE,将精液样品与各个浓度组依次混合后放置室温平衡,随即转入17 ℃电子恒温冰箱进行常温保存。全自动精子分析仪（CASA）测定1—5 d精子运动学参数（总活率与渐进性活力）,筛选出最佳适宜浓度为0.06 g·L-1。其次,将0.06 g·L-1 CAPE与400 μmol·L-1的H2O2建立氧化胁迫模型,在第1、3、5天分别采用CASA测定精子总活率与渐进性活力,荧光探针技术评估质膜完整性与顶体完整性,抗氧化试剂盒检测过氧化氢酶（catalase,CAT）与超氧化物歧化酶（superoxide dismutase,SOD）活性,在第5 天利用实时荧光定量 PCR技术测定AMPK/FOXO3a信号通路下游靶向基因（AMPK、FOXO3a、SOD1、SOD2、CAT）及凋亡基因BAX的mRNA表达水平,蛋白免疫印记技术测定AMPK、p-AMPK蛋白表达。【结果】（1）浓度筛选试验中,0.06 g·L-1的CAPE在保存第3 天显著提高精子的总活率并维持至第5天（P<0.05）,不仅如此,精子的渐进性活力在第1 天显著高于其他处理组（P<0.05）。（2）氧化胁迫试验中,H2O2处理组的精子质膜完整性、顶体完整性、总活率、渐进性活力、SOD与CAT活性在保存的第5天均极限显著低于空白组与CAPE+H2O2混合组（P<0.001）,而空白组与CAPE+H2O2混合组的总活率、顶体完整性、CAT与SOD活性不存在显著差异（P>0.05）。与CAPE+H2O2混合组相比,H2O2处理组AMPK、FOXO3a、SOD1、SOD2、CAT的mRNA表达水平显著降低（P<0.05）,BAX的表达水平显著升高（P<0.05）,而空白组与CAPE+H2O2混合组的AMPK、SOD1、CAT与BAX不存在显著差异（P>0.05）。在蛋白表达水平中,CAPE+H2O2混合组与H2O2组相比,AMPK、p-AMPK 与p-AMPK/AMPK比率显著提升（P <0.05）。【结论】在常温基础稀释液中添加0.06 g·L-1CAPE可通过促进磷酸化AMPK的表达,诱导AMPK/FOXO3a信号下游抗氧化分子的转录,继而缓解H2O2介导的氧化危害对精液品质产生的影响,延长精液的保存时间,但CAPE保护精子氧化损伤更为深入的分子机制仍需作进一步探究。 相似文献
7.
AMPK对动物营养代谢的调节作用 总被引：4，自引：0，他引：4
AMPK是一种能被腺苷一磷酸(AMP)激活的蛋白激酶。该酶的作用底物包括β—羟基—β甲基—戊二酸单酰CoA还原酶(HMGR) ,乙酰辅酶A羧化酶(ACC) ,激素敏感脂酶(HSL) ,糖原合成酶。当细胞能量被耗竭、腺苷一磷酸/腺苷三磷酸比(AMP/ATP)增加时 ,AMPK被活化 ,继而通过调节上述底物酶而改变脂类和碳水化合物代谢 ,使其朝着抑制ATP消耗过程、促进ATP生成反应的方向进行 ,从而使细胞能量得到迅速恢复。AMPK的这种“燃料开关“作用在动物抵御和适应环境应激的过程中起着重要作用。AMPK可能参与家畜应激营养代谢及营养代谢疾病的调节 相似文献
8.
AMPK参与了运动中骨骼肌葡萄糖、糖原、脂肪酸及蛋白质等主要能源物质的代谢调节,其调节机制涉及数十种下游靶。AMPK能促使GLUT4转位入肌膜,也可调节GLUT4基因表达而促进葡萄糖的摄取和氧化;抑制糖原合成酶活性及激活磷酸果糖激酶而抑制肌糖原合成和促进分解;磷酸化并灭活ACCβ及激活MCD促进脂肪酸氧化;抑制mTOR信号通路和eEF2在翻译水平上抑制蛋白质合成。 相似文献
9.
WANG Qi WANG Changjian WEI Zongyou LU Hanxi YAO Xiaolei YANG Hua WANG Feng ZHANG Yanli 《畜牧兽医学报》1956,51(12):3033-3045
This study aimed to investigate the effects of AMPK activators metformin (Met) and acadesine (AICAR) on sheep semen cryopreservation. Firstly, Met and AICAR with different concentrations (0, 100, 200, 300, 400, 500 μmol·L-1) were added into the frozen diluent. After freezing and thawing, the optimal concentration (400 μmol·L-1 Met, 200 μmol·L-1 AICAR) were selected based on sperm motility, motion performance and membrane integrity; Secondly, the collected semen was froze using different frozen diluents (control group:diluent; Met group:diluent + 400 μmol·L-1 Met; AICAR group:diluent + 200 μmol·L-1 AICAR). After thawing, the sperm AMPK protein expression, acrosomal enzyme activity, metabolic index, mitochondrial function and antioxidant enzyme activity were examined. The results showed that the addition of 400 μmol·L-1 Met and 200 μmol·L-1 AICAR could significantly improve sperm motility, motion performance and membrane integrity(P<0.05) after thawing. In 400 μmol·L-1 Met group, the total sperm motility, acrosome integrity rate and plasma membrane integrity rate were 43.20%, 91% and 46%, respectively. Compared with the control group, the level of AMPK phosphorylation in the sperm of the Met and AICAR groups was significantly increased after thawing (P<0.05), the acrosome enzyme activity of sperm was significantly increased(P<0.05); the pyruvate level was significantly decreased (P<0.05), the lactate dehydrogenase activity, lactic acid content, and ATP content were significantly increased(P<0.05). Compared with the control group, the dilution of Met group and AICAR group was more conducive to maintain mitochondrial membrane potential (P<0.05), increase ATPase and antioxidant enzyme activity (P<0.05). The addition of appropriate concentrations of Met and AICAR could improve semen quality of cryopreservation in sheep. 相似文献
10.
Lakshi A. Dayarathne Sachithra S. Ranaweera Premkumar Natraj Priyanka Rajan Young Jae Lee Chang-Hoon Han 《Journal of veterinary science (Suw?n-si, Korea)》2021,22(6)
BackgroundNaringin and its aglycone naringenin are citrus-derived flavonoids with several pharmacological effects. On the other hand, the mechanism for the anti-diabetic effects of naringenin and naringin are controversial and remain to be clarified further.ObjectiveThis study examined the relationship between glucose uptake and AMP-activated protein kinase (AMPK) phosphorylation by naringenin and naringin in high glucose-treated HepG2 cells.MethodsGlucose uptake was measured using the 2-NBDG fluorescent D-glucose analog. The phosphorylation levels of AMPK and GSK3β (Glycogen synthase kinase 3 beta) were observed by Western blotting. Molecular docking analysis was performed to evaluate the binding affinity of naringenin and naringin to the γ-subunit of AMPK.ResultsThe treatment with naringenin and naringin stimulated glucose uptake regardless of insulin stimulation in high glucose-treated HepG2 cells. Both flavonoids increased glucose uptake by promoting the phosphorylation of AMPK at Thr172 and increased the phosphorylation of GSK3β. Molecular docking analysis showed that both naringenin and naringin bind to the γ-subunit of AMPK with high binding affinities. In particular, naringin showed higher binding affinity than the true modulator, AMP with all three CBS domains (CBS1, 3, and 4) in the γ-subunit of AMPK. Therefore, both naringenin and naringin could be positive modulators of AMPK activation, which enhance glucose uptake regardless of insulin stimulation in high glucose-treated HepG2 cells.ConclusionsThe increased phosphorylation of AMPK at Thr172 by naringenin and naringin might enhance glucose uptake regardless of insulin stimulation in high glucose treated HepG2 cells. 相似文献