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腺苷2a受体抑制剂抑制人非小细胞肺癌细胞系A549增殖和迁移 预览
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作者 邵松军 段玲弟 +3 位作者 方海燕 袁国航 郭兵 张湘燕 《基础医学与临床》 CSCD 2019年第1期37-41,共5页
目的探讨腺苷2a受体抑制剂(A2aRi)对人非小细胞肺癌细胞系A549增殖、迁移和凋亡的影响及其机制。方法将细胞分为对照组及干预组,干预组予不同浓度的A2aRi处理;用MTT法检测细胞增殖;划痕实验检测细胞迁移;24h后,免疫荧光观察A2aR和caspa... 目的探讨腺苷2a受体抑制剂(A2aRi)对人非小细胞肺癌细胞系A549增殖、迁移和凋亡的影响及其机制。方法将细胞分为对照组及干预组,干预组予不同浓度的A2aRi处理;用MTT法检测细胞增殖;划痕实验检测细胞迁移;24h后,免疫荧光观察A2aR和caspase3在细胞中表达定位;用Western blot和RT-qPCR检测A2aR、caspase3蛋白和A2aRmRNA的表达。结果A2aRi能抑制A549细胞的增殖(P<0.01),其迁移能力减弱,A2aR红色荧光强度较对照组减弱,caspase3绿色荧光强度较对照组增强,两者均在胞质中表达。Caspase3蛋白表达增加(P<0.01),A2aR蛋白表达减少(P<0.01);A2aRmRNA表达下调(P<0.05)。结论A2aR在人非小细胞肺癌细胞系A549中高表达,A2aRi能抑制A549细胞增殖和减弱其迁移能力。 展开更多
关键词 非小细胞肺癌 腺苷 腺苷受体 CASPASE3
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替格瑞洛影响冠心病患者血浆腺苷浓度的研究进展
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作者 徐汝明 马丽萍 《第二军医大学学报》 CAS CSCD 北大核心 2019年第6期664-667,共4页
替格瑞洛是一种新型口服抗血小板药物,通过直接作用于二磷酸腺苷(ADP)受体P2Y12抑制ADP介导的血小板聚集,发挥抗血小板功能。除抑制血小板聚集外,替格瑞洛还表现出生物多效性,如增加冠状动脉血流、限制心肌梗死面积和诱发呼吸困难等。... 替格瑞洛是一种新型口服抗血小板药物,通过直接作用于二磷酸腺苷(ADP)受体P2Y12抑制ADP介导的血小板聚集,发挥抗血小板功能。除抑制血小板聚集外,替格瑞洛还表现出生物多效性,如增加冠状动脉血流、限制心肌梗死面积和诱发呼吸困难等。研究认为替格瑞洛的生物多效性与腺苷密切相关,但目前有关替格瑞洛对血浆腺苷浓度影响的研究结果并不一致,可能与血浆腺苷浓度的不同测定方法(尤其是终止液配比)、不同测定时间点等有关。规范血浆腺苷浓度的测定方法以及动态监测服用替格瑞洛后血浆腺苷浓度的变化,有助于进一步明确替格瑞洛对血浆腺苷浓度的影响。 展开更多
关键词 冠心病 替格瑞洛 腺苷 血浆浓度
常压室温等离子体和紫外诱变选育腺苷高产菌株 预览
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作者 袁红梅 薛正莲 杨心萍 《发酵科技通讯》 CAS 2019年第2期100-105,共6页
腺苷是一种内源性核苷类物质,在生理生化过程中起着重要的调控作用。使用常压室温等离子体(ARTP)诱变、UV-LiCl诱变和复合诱变,将诱变后的菌株涂布在含最低致死浓度的6-巯基嘌呤(6-MP)抗性平板和不含该抗性的平板上,35℃培养1~2d后对其... 腺苷是一种内源性核苷类物质,在生理生化过程中起着重要的调控作用。使用常压室温等离子体(ARTP)诱变、UV-LiCl诱变和复合诱变,将诱变后的菌株涂布在含最低致死浓度的6-巯基嘌呤(6-MP)抗性平板和不含该抗性的平板上,35℃培养1~2d后对其抗性正突变株进行发酵筛选和传代稳定性考察,获得1株腺苷高产菌株A-Z-L-2,传代6次后其腺苷产量达到20.512g/L,较出发菌株BS-0腺苷产量提高了16.12%,且该菌株具有较好的传代稳定性。 展开更多
关键词 腺苷 枯草芽孢杆菌 6-巯基嘌呤 常压室温等离子体 紫外 正突变
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Intraperitoneal injection adenovirusPmultirnai-ldhcOkay Plateau pika (Ochotona curzoniae) Skeletal MuscleLdhcGene of Silence effect 预览
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作者 Yangzhi Deng 《探索-植物与动物科学》 2019年第1期25-29,共5页
Intraperitoneal injection is a kind of simple and convenient to drug of style in order to verify intraperitoneal injection adenovirusPmultirnai-ldhcThe plateau pika (Ochotona curzoniae (Ochotona curzoniac) Skeletal Mu... Intraperitoneal injection is a kind of simple and convenient to drug of style in order to verify intraperitoneal injection adenovirusPmultirnai-ldhcThe plateau pika (Ochotona curzoniae (Ochotona curzoniac) Skeletal MuscleLdhcGene Silence of feasibility will27Only plateau pika (Ochotona curzoniae) divided into interference group, Shell Group and blank control group each group9Only individual interference group and Shell Group respectively Injection0.65 mLAdenovirusPmultirnai-ldhcAnd adenovirusPmultirnai-NSBlank control group injected with the same amount of physiological saline injection after7/dDetection Skeletal Muscle inLdhcGeneMRNAAnd protein of Expression Level Determination the lactate dehydrogenase (LDH) Activity and lactic acid (LD) And three phosphoric acid adenosine (ATP) Of content. Results show that and blank control group compared interference group inMRNAAND PROTEIN LEVEL,LdhcGene expression respectively reduce41.73%And15.76%;Lactate dehydrogenase (LDH) Activity, lactic acid (LD) AndATPContent respectively reduce23.98%,51.08%And19.29%. Results (intraperitoneal injection adenovirusPmultirnai-ldhcCan effective silence Skeletal Muscle inLdhcGene expression. 展开更多
关键词 INTRAPERITONEAL injection plateau pika LACTATE dehydrogenase LACTIC ACID three phosphoric ACID adenosine
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UPLC-MS/MS同时测定熟地黄中8个核苷类成分的含量
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作者 张艳丽 杨雁芸 +5 位作者 白志尧 曹彦刚 孙亚萍 刘通 克迎迎 吴亚 《药物分析杂志》 CAS CSCD 北大核心 2019年第4期608-614,共7页
目的:建立超高效液相色谱-三重四极杆质谱法(UPLC-MS/MS法)同时测定熟地黄中鸟苷、肌苷、尿苷、尿嘧啶、腺苷、胞苷、腺嘌呤、次黄嘌呤8个核苷的含量。方法:采用ACQUITY UPLC BEH C18(2.1 mm×50 mm,1.7μm)色谱柱,以0.1%甲酸溶液-... 目的:建立超高效液相色谱-三重四极杆质谱法(UPLC-MS/MS法)同时测定熟地黄中鸟苷、肌苷、尿苷、尿嘧啶、腺苷、胞苷、腺嘌呤、次黄嘌呤8个核苷的含量。方法:采用ACQUITY UPLC BEH C18(2.1 mm×50 mm,1.7μm)色谱柱,以0.1%甲酸溶液-甲醇为流动相梯度洗脱,流速0.2 mL·min-1,柱温30℃,进样量2μL,ESI+,多反应监测(MRM)对熟地黄饮片中8个核苷类成分进行含量测定。结果:所测8个核苷类成分在测定质量浓度范围内线性关系良好(R2>0.996 0),平均加样回收率为98.6%~102.0%,RSD为1.3%~2.9%。6批样品中鸟苷、肌苷、尿苷、尿嘧啶、腺苷、胞苷、腺嘌呤、次黄嘌呤的含量范围分别为0.045~71.838、3.220~11.253、84.613~102.508、1.516~4.893、6.426~68.003、0.138~17.437、0.423~1.953、0.142~1.216μg·g-1。结论:该方法适用于同时测定熟地黄饮片中鸟苷、肌苷、尿苷、尿嘧啶、腺苷、胞苷、腺嘌呤、次黄嘌呤8个核苷类成分含量,可用于熟地黄饮片的质量控制。 展开更多
关键词 熟地黄 核苷 鸟苷 肌苷 尿苷 尿嘧啶 腺苷 胞苷 腺嘌呤 次黄嘌呤 含量测定 质量控制 超高效液相色谱-串联质谱
HILIC-MS/MS技术同时检测蛹虫草中虫草素、虫草酸、腺苷和肌苷含量的方法学 预览
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作者 张铭雅 朱志铭 +5 位作者 姚迪 陈俏慧 鲍晓文 陈嘉伟 朱健伟 马博 《生物加工过程》 CAS 2019年第4期430-436,共7页
蛹虫草是我国具有极高药用价值的滋补中药材。对蛹虫草成分的含量进行准确的定性定量分析,可为相关产品质量控制、筛选高活性蛹虫草及进一步研究蛹虫草的功效提供技术手段。建立一种灵敏稳定的亲水作用色谱法串联质谱(HILICMS/MS)方法,... 蛹虫草是我国具有极高药用价值的滋补中药材。对蛹虫草成分的含量进行准确的定性定量分析,可为相关产品质量控制、筛选高活性蛹虫草及进一步研究蛹虫草的功效提供技术手段。建立一种灵敏稳定的亲水作用色谱法串联质谱(HILICMS/MS)方法,同时对蛹虫草中虫草素、虫草酸(D甘露糖醇)、腺苷和肌苷的含量进行测定。研究发现其线性范围分别为0.02~5μg/mL(R^2=0.9953)、0.2~50μg/mL(R^2=0.9994)、0.002~0.5μg/mL(R^2=0.9970)和0.002~0.5μg/mL(R^2=0.9950)。采用HILICMS/MS技术,测定结果精密度小于10.0%,准确度在8.0%左右,具有较好的特异性、灵敏度和重现性。 展开更多
关键词 蛹虫草 HILIC-MS/MS 虫草素 虫草酸 腺苷
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Acute drivers of neuroinflammation in traumatic brain injury 预览
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作者 Kathryn L.Wofford David J.Loane D.Kacy Cullen 《中国神经再生研究:英文版》 SCIE CAS CSCD 2019年第9期1481-1489,共9页
Neuroinflammation is initiated as a result of traumatic brain injury and can exacerbate evolving tissue pathology.Immune cells respond to acute signals from damaged cells,initiate neuroinflammation,and drive the patho... Neuroinflammation is initiated as a result of traumatic brain injury and can exacerbate evolving tissue pathology.Immune cells respond to acute signals from damaged cells,initiate neuroinflammation,and drive the pathological consequences over time.Importantly,the mechanism(s)of injury,the location of the immune cells within the brain,and the animal species all contribute to immune cell behavior following traumatic brain injury.Understanding the signals that initiate neuroinflammation and the context in which they appear may be critical for understanding immune cell contributions to pathology and regeneration.Within this paper,we review a number of factors that could affect immune cell behavior acutely following traumatic brain injury. 展开更多
关键词 traumatic BRAIN INJURY inflammation NEUROINFLAMMATION MICROGLIA macrophage ACUTE diffuse BRAIN INJURY cytokines ADENOSINE 5′-triphosphoate glutamate calcium
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Long-term adenosine A1 receptor activation-induced sortilin expression promotesα-synuclein upregulation in dopaminergic neurons 预览
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作者 Yun-Cheng Lv An-Bo Gao +7 位作者 Jing Yang Li-Yuan Zhong Bo Jia Shu-Hui Ouyang Le Gui Tian-Hong Peng Sha Sun Francisco S.Cayabyab 《中国神经再生研究:英文版》 SCIE CAS CSCD 2020年第4期712-723,共12页
Prolonged activation of adenosine A1 receptor likely leads to damage of dopaminergic neurons and subsequent development of neurodegenerative diseases.However,the pathogenesis underlying long-term adenosine A1 receptor... Prolonged activation of adenosine A1 receptor likely leads to damage of dopaminergic neurons and subsequent development of neurodegenerative diseases.However,the pathogenesis underlying long-term adenosine A1 receptor activation-induced neurodegeneration remains unclear.In this study,rats were intraperitoneally injected with 5 mg/kg of the adenosine A1 receptor agonist N6-cyclopentyladenosine(CPA)for five weeks.The mobility of rats was evaluated by forced swimming test,while their cognitive capabilities were evaluated by Y-maze test.Expression of sortilin,α-synuclein,p-JUN,and c-JUN proteins in the substantia nigra were detected by western blot analysis.In addition,immunofluorescence staining of sortilin andα-synuclein was performed to detect expression in the substantia nigra.The results showed that,compared with adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine(5 mg/kg)+CPA co-treated rats,motor and memory abilities were reduced,surface expression of sortin andα-synuclein in dopaminergic neurons was reduced,and total sortilin and totalα-synuclein were increased in CPA-treated rats.MN9D cells were incubated with 500 nM CPA alone or in combination with 10μM SP600125(JNK inhibitor)for 48 hours.Quantitative real-time polymerase chain reaction analysis of sortilin andα-synuclein mRNA levels in MN9D cells revealed upregulated sortilin expression in MN9D cells cultured with CPA alone,but the combination of CPA and SP600125 could inhibit this expression.Predictions made using Jasper,PROMO,and Alibaba online databases identified a highly conserved sequence in the sortilin promoter that was predicted to bind JUN in both humans and rodents.A luciferase reporter assay of sortilin promoter plasmid-transfected HEK293T cells confirmed this prediction.After sortilin expression was inhibited by sh-SORT1,expression of p-JUN and c-JUN was detected by western blot analysis.Long-term adenosine A1 receptor activation levels upregulatedα-synuclein expression at the post-transcriptional level by affecting sort 展开更多
关键词 cognitive dysfunction dopaminergic neuron DYSKINESIA JNK/c-JUN pathway long-term adenosine A1 receptor activation neural regeneration neurodegenerative diseases SORTILIN Α-SYNUCLEIN
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腺苷对肺癌脑转移的影响及其可能机制 预览
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作者 唐明科 魏殿芳 +2 位作者 刘洋 张超越 秦丽娟 《临床与实验病理学杂志》 CAS CSCD 北大核心 2019年第1期19-22,共4页
目的探讨腺苷对肺癌脑转移的影响及其可能机制。方法采用Western blot法检测肺癌细胞内缺氧诱导因子-1(hypoxia inducible factor-1,HIF-1)和血脑屏障脑微血管内皮细胞上的紧密连接蛋白ZO-1的表达水平;应用ELISA法测定肺癌细胞培养液内... 目的探讨腺苷对肺癌脑转移的影响及其可能机制。方法采用Western blot法检测肺癌细胞内缺氧诱导因子-1(hypoxia inducible factor-1,HIF-1)和血脑屏障脑微血管内皮细胞上的紧密连接蛋白ZO-1的表达水平;应用ELISA法测定肺癌细胞培养液内腺苷的含量。采用荧光分析法检测体外血脑屏障模型的通透性改变;用Hemocytometer计数Transwell下室的A549肺癌细胞数。结果肺癌细胞中HIF-1的表达和肺癌细胞培养液内腺苷含量均于肺癌细胞缺氧12 h时达最高水平。与此同时,血脑屏障ZO-1蛋白的表达最低,血脑屏障通透性最大(7.11),透过血脑屏障模型的肺癌细胞数最多(84.6)。腺苷引起血脑屏障通透性增加,其变化趋势与缺氧肺癌细胞培养液的作用一致。结论缺氧可引起肺癌细胞释放腺苷增多,增多的腺苷导致血脑屏障紧密连接蛋白ZO-1表达减少,导致血脑屏障通透性增大,最终引起肺癌脑转移。 展开更多
关键词 肺肿瘤 脑转移 腺苷 HIF-1 脑微血管内皮细胞 ZO-1蛋白
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Preventive Effect of Perioperative Management Using Oral Clopidogrel on Postoperative Rebleeding in Patients with Spontaneous Intracerebral Hemorrhage 预览
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作者 Tao Yang Zhong-Xiao Lin +1 位作者 Xi-Lin Zhang Xing-Ming Zhong 《脑血管病电子杂志(英文)》 2019年第1期11-14,共4页
Background: With the aging society in China and the increasing incidence of cardiovascular and cerebrovascular diseases,spontaneous cerebral hemorrhage with the use of oral clopidogrel has become common in patients. O... Background: With the aging society in China and the increasing incidence of cardiovascular and cerebrovascular diseases,spontaneous cerebral hemorrhage with the use of oral clopidogrel has become common in patients. Owing to its inhibition of platelet aggregation,oral clopidogrel results in a high incidence of postoperative intracranial rebleeding,and the prognosis is poor. The purpose of this study is to determine the effect of perioperative management of spontaneous intracerebral hemorrhage (SICH) using oral clopidogrel on postoperative rebleeding. Methods: From January 2010 to June 2018,64 patients with SICH who took clopidogrel orally and finally received surgical treatment were selected from Huzhou First People’s Hospital and the Second Affiliated Hospital of Wenzhou Medical University. The influencing factors of postoperative rebleeding were analyzed via a case-control study. Results: All patients with SICH were divided into two groups according to the standard time of drug withdrawal before surgery: drug withdrawal at < 3 days and ≥ 3 days. The incidence of postoperative rebleeding after drug withdrawal at ≥ 3 days was significantly lower than that after drug withdrawal at < 3 days. Thereafter,all patients were divided into two groups according to the adenosine diphosphate (ADP) inhibition rate detected using a thromboelastography monitor: ADP inhibition rate of < 60% and ≥ 60%. The incidence of postoperative rebleeding in the ADP inhibition rate of < 60% group was significantly lower than that in the ADP inhibition rate of ≥ 60% group. In addition,drug withdrawal at ≥ 3 days and ADP inhibition rate of < 60% had a synergistic effect on the prevention of postoperative rebleeding. Conclusions: Patients with drug withdrawal at ≥ 3 days can undergo active early surgery. The ADP inhibition rate of ≥ 60% can be used as the predictive index of high-risk postoperative rebleeding in patients with SICH taking oral clopidogrel. By reducing the ADP inhibition rate to < 60%,the incidence of postoperativ 展开更多
关键词 Spontaneous INTRACEREBRAL HEMORRHAGE CLOPIDOGREL ADENOSINE DIPHOSPHATE PERIOPERATIVE management
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HPLC法同时测定滋阴温阳活血颗粒中两种指标性成分的研究
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作者 文小燕 谭梅英 +1 位作者 容丽英 陈伟英 《湖南中医杂志》 2019年第6期139-141,共3页
目的:建立滋阴温阳活血颗粒中两种指标性成分(腺苷和山奈酚-3-O-β-D-葡萄糖苷)的同时测定方法。方法:采用HPLC法,选用phenomex Kinetex C18色谱柱(4. 6mm×250mm,5μm),0. 1%磷酸-乙腈为流动相,梯度洗脱,流速1. 0ml/min,柱温30℃,... 目的:建立滋阴温阳活血颗粒中两种指标性成分(腺苷和山奈酚-3-O-β-D-葡萄糖苷)的同时测定方法。方法:采用HPLC法,选用phenomex Kinetex C18色谱柱(4. 6mm×250mm,5μm),0. 1%磷酸-乙腈为流动相,梯度洗脱,流速1. 0ml/min,柱温30℃,进样量10μm,检测波长254nm;以腺苷和山奈酚-3-O-β-D-葡萄糖苷为指标性成分,共测定10批滋阴温阳活血颗粒。结果:10批颗粒中,腺苷的含量在2. 706~4. 132mg/g范围内,山奈酚-3-O-β-D-葡萄糖苷的含量在3. 692~4. 948mg/g范围内。结论:本研究所建立方法快速、准确、高效,能够对滋阴温阳活血颗粒的质量进行科学评价。 展开更多
关键词 滋阴温阳活血颗粒 腺苷 山奈酚-3-O-β-D-葡萄糖苷 含量测定
针刺预处理对心肌缺血再灌注大鼠Akt/eNOS通路的影响 预览
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作者 徐文博 唐关敏 +4 位作者 钱钢 胡惠林 汪世军 金丹丹 翟昌林 《中国中医药科技》 CAS 2019年第1期5-7,114共4页
目的:研究针灸预处理通过激活Akt/eNOS通路对大鼠心肌缺血再灌注的保护作用。方法:将40只雄性SD大鼠随机分为4组,假手术组(Sham组)、缺血再灌注组(I/R组)、针刺预处理组(AP组)、针刺对照组(AC组),其中针刺预处理组造模前7 d即每天给予电... 目的:研究针灸预处理通过激活Akt/eNOS通路对大鼠心肌缺血再灌注的保护作用。方法:将40只雄性SD大鼠随机分为4组,假手术组(Sham组)、缺血再灌注组(I/R组)、针刺预处理组(AP组)、针刺对照组(AC组),其中针刺预处理组造模前7 d即每天给予电针"内关穴"处理,而AC组则予电针"外关穴"处理,建立心肌缺血再灌注模型,心脏压力导管检测系统测定各组大鼠心功能指标,比色法测定血清腺苷浓度,Western blot检测心肌细胞Akt/eNOS通路的活化情况。结果:与I/R组比较,AP组心功能明显改善,Akt/eNOS信号通路磷酸化水平显著增加(P <0. 01),血清腺苷含量明显提升(P <0. 01);而AC组与I/R组比较心功能改善不明显,Akt/eNOS、腺苷的表达比较均无统计学意义(均P> 0. 05)。结论:针灸"内关穴"预处理可通过激活Akt/eNOS通路发挥心肌缺血再灌注损伤的保护作用,其机制可能与激活腺苷表达相关。 展开更多
关键词 心肌缺血再灌注 针刺预处理 内关穴 Akt/eNOS 腺苷 大鼠
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腺苷的主要功能和发酵法生产研究进展 预览
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作者 袁红梅 薛正莲 +3 位作者 杨心萍 宋词 章杰 张伟豪 《发酵科技通讯》 CAS 2019年第3期150-153,共4页
腺苷是一种具有重要生理生化功能的核苷类物质,可以合成ATP,ADP,cAMP,SAM,阿糖腺苷和嘌呤霉素等物质,还可以直接作用于心脑血管、中枢神经、肾脏和肺等系统。综述了腺苷的功能和腺苷的微生物发酵生产途径和育种思路,讨论了发酵法生产腺... 腺苷是一种具有重要生理生化功能的核苷类物质,可以合成ATP,ADP,cAMP,SAM,阿糖腺苷和嘌呤霉素等物质,还可以直接作用于心脑血管、中枢神经、肾脏和肺等系统。综述了腺苷的功能和腺苷的微生物发酵生产途径和育种思路,讨论了发酵法生产腺苷的意义及存在的问题。生物法中微生物发酵法生产腺苷成本低、条件温和、提取率高,对环境几乎不造成危害,是今后腺苷生产的主要方法。 展开更多
关键词 腺苷 功能 合成 发酵 代谢
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基于石墨烯的电化学基因传感器
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作者 张韩洁 侯晓琳 +3 位作者 郑梅琼 连玲娜 龚微微 杨剑萍 《分析试验室》 CAS CSCD 北大核心 2019年第4期489-494,共6页
石墨烯及其复合物是构建新型电化学生物传感器的理想材料,为临床诊断、食品安全、环境监测等研究领域提供了材料基础。本文简述了几种有效的石墨烯制备方法:剥离法、化学气相沉积法、固态碳源催化法和氧化还原法等;论述了基于石墨烯的... 石墨烯及其复合物是构建新型电化学生物传感器的理想材料,为临床诊断、食品安全、环境监测等研究领域提供了材料基础。本文简述了几种有效的石墨烯制备方法:剥离法、化学气相沉积法、固态碳源催化法和氧化还原法等;论述了基于石墨烯的电化学基因生物传感器的应用研究,如在脱氧核糖核酸(DNA)、核酸类似物(PNA)、核糖核酸(RNA)和腺苷的检测应用。本文为基于石墨烯的生物传感器的构建提供了纳米材料制备方法和生物传感策略等方面的参考,并展望了基因生物传感器的未来发展及应用方向。 展开更多
关键词 石墨烯 脱氧核糖核酸 核糖核酸 腺苷 生物传感器
Ginsenoside Rb1 Ameliorates Autophagy of Hypoxia Cardiomyocytes from Neonatal Rats via AMP-Activated Protein Kinase Pathway
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作者 DAI Sheng-nan HOU Ai-jie +4 位作者 ZHAO Shu-mei CHEN Xiao-ming HUANG Hua-ting CHEN Bo-han KONG Hong-liang 《中国结合医学杂志:英文版》 SCIE CAS CSCD 2019年第7期521-528,共8页
Objective: To investigate whether ginsenoside-Rb1(Gs-Rb1) improves the CoCl2-induced autophagy of cardiomyocytes via upregulation of adenosine 5’-monophosphate-activated protein kinase(AMPK) pathway. Methods: Ventric... Objective: To investigate whether ginsenoside-Rb1(Gs-Rb1) improves the CoCl2-induced autophagy of cardiomyocytes via upregulation of adenosine 5’-monophosphate-activated protein kinase(AMPK) pathway. Methods: Ventricles from 1-to 3-day-old Wistar rats were sequentially digested, separated and incubated in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum for 3 days followed by synchronization. Neonatal rat cardiomyocytes were randomly divided into 7 groups: control group(normal level oxygen), hypoxia group(500 μmol/L CoCl2), Gs-Rb1 group(200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2), Ara A group(500 μmol/L Ara A + 500 μmol/L CoCl2), Ara A+ Gs-Rb1 group(500 μmol/L Ara A + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2), AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide(AICAR)+ 500 μmol/L CoCl2], and AICAR+Gs-Rb1 group(1 mmol/L AICAR + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl2). Cel s were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay and cardiac troponin I(cTnI) levels were detected by enzyme-linked immunosorbent assay(ELISA). AMPK activity was assessed by 2’,7’-dichlorofluorescein diacetate(DCFH-DA) ELISA assay. The protein expressions of Atg4 B, Atg5, Atg6, Atg7, microtubule-associated protein 1 A/1 B-light chain 3(LC3), P62, and active-cathepsin B were measured by Western blot. Results: Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes(P<0.01). However, the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A(P<0.01). Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes. The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A(P<0.01) and was not affected by AICAR(P=0.983). Gs-Rb1 up-regulated Atg4B, Atg5, Beclin-1, Atg7, LC3B Ⅱ, the LC3BⅡ/Ⅰ ratio and cathepsin B activity of hypoxia cardiomyocytes(P<0.05), each of these protein levels was significantly enhanced by Ara A(all P<0.01), but was not affected by AICAR(all P 展开更多
关键词 CARDIOMYOCYTES ginsenosides-Rb1 HYPOXIA ADENOSINE 5'-monophosphate-activated protein KINASE autophagic flux
腺苷负荷对健康小鼠心肌灌注的影响
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作者 黄凌霙 邓又斌 +6 位作者 刘娅妮 王译斌 张隽 田洁 王佳玉 毛宇航 王心宇 《中华超声影像学杂志》 CSCD 北大核心 2019年第5期444-450,共7页
目的应用心肌造影超声心动图(myocardial contrast echocardiography,MCE)评价腺苷负荷前后小鼠心肌灌注的变化。方法采用Vevo2100型高分辨小动物超声仪,在静息状态下和静脉输注腺苷后对26只健康C57BL /6小鼠行心肌造影超声心动图检查... 目的应用心肌造影超声心动图(myocardial contrast echocardiography,MCE)评价腺苷负荷前后小鼠心肌灌注的变化。方法采用Vevo2100型高分辨小动物超声仪,在静息状态下和静脉输注腺苷后对26只健康C57BL /6小鼠行心肌造影超声心动图检查。连续获取小鼠胸骨旁左心长轴切面和乳头肌短轴切面动态图像。所有动态图像存盘以脱机分析,得到静息及腺苷负荷状态下左室心肌的灌注定量参数。结果 26只实验动物成功完成腺苷负荷检查。常规超声心动图获取小鼠心率以及左室各结构与功能参数,静息与负荷状态下,以上参数差异无统计学意义(P>0.05);腺苷负荷状态下,左室长轴切面前间隔及后壁中间段、乳头肌短轴切面前壁、侧壁、下壁和间隔壁上升期造影剂强度随时间变化的斜率(β值)较静息状态下明显升高,差异具有统计学意义(均P<0.05);心肌各节段平台期造影剂的峰值强度(A值)则未见明显增加(均P>0.05);心肌各节段局部心肌血流量(myocardial blood flow,MBF)较静息状态下明显增加(均P<0.05)。结论初步得到小鼠腺苷负荷前后心肌血流灌注的生理变化特点,为今后腺苷负荷超声心动图在小鼠冠心病等心血管疾病模型中的应用提供了依据。 展开更多
关键词 超声心动描记术 压力 腺苷 小鼠 心肌灌
β-Cyclodextrin polymer based fluorescence enhancement method for sensitive adenosine triphosphate detection
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作者 Chunxia Song Yuxin Xiao +2 位作者 Kunpeng Li Xiaoyu Zhang Ying Lu 《中国化学快报:英文版》 SCIE CAS CSCD 2019年第6期1249-1252,共4页
Adenosine triphosphate plays a crucial role in regulation of many biological pathways and has been used as an indicator for many diseases.In this pape r,based on the fact that β-cyclodextrin polymer(polyβ-CD)could s... Adenosine triphosphate plays a crucial role in regulation of many biological pathways and has been used as an indicator for many diseases.In this pape r,based on the fact that β-cyclodextrin polymer(polyβ-CD)could significantly enhance pyrene fluorescence through supramolecular assembly(host-gest interaction),a sensitive and facile method for adenosine triphosphate detection has been developed.A 3’-pyrene-labelled ATP-binding aptamer was employed as the fluorescence probe,which could be digested by exonuclease I to obtain mononucleotides,with pyrene attached on.The pyrene attached on mononucleotides could easily enter the hydrophobic cavity of polyβ-CD,accompanied with prominent fluo rescence enhancement.While ATP was introduced,ATP and its aptamer could combine together and the obtained hairpin complex could not be cleaved by exonuclease Ⅰ,The pyrene labelled on the probe could not enter the cavity of polyβ-CD belong to the complex’steric hindrance,accompanied with the weak pyrene’fluorescence.So we could quantify the concentration of adenosine triphosphate facilely by measuring the fluorescence intensity of the system.The detection limit of this method for adenosine triphosphate was 11 μmol/L(S/N=3),The developed method showed sufficient selectivity and could successfully assay adenosine triphosphate in biological samples.The developed method provides a potential platform for biological micromoles assay. 展开更多
关键词 Β-CYCLODEXTRIN polymer SUPRAMOLECULAR assembly FLUORESCENCE Enhancement ADENOSINE TRIPHOSPHATE
Protein kinase A inhibition induces EPAC-dependent acrosomal exocytosis in human sperm
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作者 Diana Itzhakov Yeshayahu Nitzan Haim Breitbart 《亚洲男性学杂志:英文版》 SCIE CAS CSCD 2019年第4期337-344,共8页
To in teract with the egg, the spermatozo on must un dergo several biochemical and motility modifications in the female reproductive tract, collectively called capacitation. Only capacitated sperm can undergo acrosoma... To in teract with the egg, the spermatozo on must un dergo several biochemical and motility modifications in the female reproductive tract, collectively called capacitation. Only capacitated sperm can undergo acrosomal exocytosis, near or on the egg, a process that allows the sperm to penetrate and fertilize the egg. In the present study, we investigated the invoIvement of cyclic adenosine monophosphate (cAMP)-dependent processes on acrosomal exocytosis. Inhibition of protein kinase A (PKA) at the end of capacitation induced acrosomal exocytosis. This process is cAMP-dependent;however, the addition of relatively high concentration of the membrane-permeable 8-bromo-cAMP (8Br-cAMP, 0.1 mmol l^-1) analog induced significant inhibition of the acrosomal exocytosis. The induction of acrosomal exocytosis by PKA inhibition was significantly inhibited by an exchange protein directly activated by cAMP (EPAC) ESI09 inhibitor. The EPAC selective substrate activated AE at relatively low concentrations (0.02-0.1 μmol l^1), whereas higher concerttrations (>5 pmol l^-1) were inhibitory to the AE induced by PKA inhibition. Inhibition of PKA revealed about 50% increase in intracellular cAMP levels, conditions under which EPAC can be activated to induce the AE. Induction of AE by activating the actin severing?protein, gelsolin, which causes F-actin dispersion, was inhibited by the EPAC inhibitor. The AE induced by PKA inhibition was mediated by phospholipase C activity but not by the Ca^2+-channel, CatSper. Thus, inhibition of PKA at the end of the capacitation process induced EPAC/phospholipase C-dependent acrosomal exocytosis. EPAC mediates F-actin depolymerization an d/or activation of effectors down stream to F-actin breakdown that lead to acrosomal exocytosis. 展开更多
关键词 acrosomal EXOCYTOSIS exchange PROTEIN directly activated by cyclic ADENOSINE MONOPHOSPHATE PROTEIN kinase A sperm
UPLC-MS/MS法同时测定感冒宁颗粒5种成分含量
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作者 柳春娣 张卫 +4 位作者 施晓卉 刘晓燕 韩怡 吕玲燕 蒋志涛 《国际中医中药杂志》 2019年第1期62-66,共5页
目的建立同时测定感冒宁颗粒中腺苷、绿原酸、咖啡酸、黄芩苷、汉黄芩素含量的方法。方法采用超高效液相色谱-串联质谱(ultra performance liquid chromatography tandem mass spectrometry, UPLC-MS/MS)联用技术,色谱柱为Agilent ZOBAX... 目的建立同时测定感冒宁颗粒中腺苷、绿原酸、咖啡酸、黄芩苷、汉黄芩素含量的方法。方法采用超高效液相色谱-串联质谱(ultra performance liquid chromatography tandem mass spectrometry, UPLC-MS/MS)联用技术,色谱柱为Agilent ZOBAX SB C18 (2.1 mm×150 mm,5 μm),流动相为甲醇-2 mmol/L乙酸铵水溶液,等度洗脱,流速0.2 ml/min,柱温35 ℃,进样量2 μl;采用电喷雾化离子源,正负离子切换模式,多反应监测模式进行定量分析,离子源温度300 ℃,干燥气温度400 ℃,干燥气流量为20 L/min,雾化气压力55 psi,毛细管电压4 000 V。结果感冒宁颗粒中腺苷、绿原酸、咖啡酸、黄芩苷、汉黄芩素检测质量浓度线性范围分别为0.20~12.80 ng (r=0.999 6)、1.00~64.00 ng (r=0.999 8)、0.40~25.60 ng (r=0.999 6)、4.00~256.00 ng (r=0.999 2)、0.20~12.8 ng (r=0.999 1),线性关系良好;检测限分别为0.02、0.10、0.04、0.40、0.02 ng,定量限分别为0.04、0.20、0.08、0.80、0.04 ng;精密度、重复性、稳定性的RSD均≤3.5%;平均加样回收率在99.30%~100.75%之间,RSD为2.09%~3.17%。结论所建立的方法简单、快速、灵敏、准确,可同时测定感冒宁颗粒中5种成分的含量,可用于感冒宁颗粒的质量控制。 展开更多
关键词 质量控制 感冒宁颗粒 超高效液相色谱-串联质谱法 腺苷 绿原酸 咖啡酸 黄芩苷 汉黄芩素
Treadmill training improves neurological deficits and suppresses neuronal apoptosis in cerebral ischemic stroke rats 预览
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作者 Li-Mei Cao Zhi-Qiang Dong +1 位作者 Qiang Li Xu Chen 《中国神经再生研究:英文版》 SCIE CAS CSCD 2019年第8期1387-1393,共7页
RehabilNation training is believed to be beneficial to patients with stroke, but its molecular mechanism is still unclear. Rat models of cerebral ischemic stroke were established by middle cerebral artery occlusion/re... RehabilNation training is believed to be beneficial to patients with stroke, but its molecular mechanism is still unclear. Rat models of cerebral ischemic stroke were established by middle cerebral artery occlusion/reperfusion, and then received treadmill training of different intens让ies, twice a day for 30 minutes for 1 week. Low-intensity training was conducted at 5 m/min, with a 10-minute running, 10-minute rest, and 10-minute running cycle. In the moderate-intensity training, the intensity gradually increased from 5 m/min to 10 m/min in 5 minutes, with the same rest cycle as above. In high-intensity training, the intensity gradually increased from 5 m/min to 25 m/min in 5 minutes, with the same rest cycle as above. The Bederson scale was used to evaluate the improvement of motor function. Infarct volume was detected using 2,3,5-triphenyltetrazolium chloride staining. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining was applied to detect the apoptosis of nerve cells in brain tissue. Western blot assay was employed to analyze the activation of cyclic adenosine monophosphate (cAMP)/protein kinase A and Akt/glycogen synthase kinase-3卩 signaling pathways in rat brain tissue. All training intensities reduced the neurological deficit score, infarct volume, and apoptosis in nerve cells in brain tissue of stroke rats. Training intensities activated the cAMP/protein kinase A and Akt/glycogen synthase kinase-3 beta signaling pathways. This activation was more obvious with higher training intensities. These changes were reversed by intracerebroventricular injection of protein kinase A inhibitor Rp-cAMP. Our findings indicate that the neuroprotective effect of rehabilitation training is achieved via activation of the cAMP/ protein kinase A and Akt/glycogen synthase kinase-3 beta signaling pathways. This study was approved by the Ethics Committee of Animal Experimentation in Shanghai No. 8 Peoples Hospital, China. 展开更多
关键词 nerve REGENERATION ischemic stroke TREADMILL training neuronal DEFICIT apoptosis cyclic adenosine MONOPHOSPHATE protein kinase A GLYCOGEN synthase kinase-3^ NEUROPROTECTIVE effect neural REGENERATION
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