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Science China(Life Sciences) 2015年第6期58卷 531-540页

作者： CHEN Li TAO Yong JIANG YanRong Department of Ophthalmology People's Hospital Peking University Key Laboratory of Vision Loss and Restoration Ministry of Education Beijing Key Laboratory of Diagnosis and Therapy of Retinal and Choroid Diseases

This paper aims to observe the changes of the inflammatory cytokines in microglial BV2 cells stimulated by apelin, and inves- tigate the mechanism of inflammatory cytokines secretion after apelin stimulation. Immunofluorescence and quantitative re- al-time PCR were performed to observe expression of TNF-α, IL-Iβ, IL-10, MIP-let, and MCP-1 in BV2 cells. Western blot was used to investigate the expression of phosphorylation PI-3K/Akt and phosphorylation Erk signaling pathways in BV2 cells after stimulation by apelin. Furthermore, PI-3K/Akt inhibitor （LY294402） and Erk inhibitor （U0126） were used as antagonists to detect the secretion mechanisms of cytokines in BV2 cells stimulated by apelin. Exogenous recombinant apelin activated the expression of TNF-α, IL-1β, MCP-1 and MIP-1α in BV2 cells by the detection of fluorescence expression and mRNA. apelin also unregulated the protein expression of p-PI-3K/Akt and p-Erk in BV2 cells induced by apelin. LY294002 and U0126 in- hibited activation of p-PI-3K/Akt and p-Erk expression by Western blot and attenuated the expression of inflammation factors in BV2 cells by fluorescence staining. This study demonstrates that apelin is a potential activator of inflammation factors through the PI3K/Akt and Erk signaling pathway and is potential therapeutically relevant to inflammatory responses of micro- glia cells.

关键词： apelin inflammation cytokines BV2 PI-3K/Akt MEK/Erk

apelin-APJ Effects of Ginsenoside-Rb1 Depending on Hypoxia-Induced Factor 1α in Hypoxia Neonatal Cardiomyocytes

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Chinese Journal of Integrative Medicine 2015年第2期21卷 139-146页

作者：孔宏亮李占全赵淑梅袁龙苗志林刘莹关汝明 Cardiology Center The People's Hospital of Liaoning Province Shenyang Normal University

Objective： To investigate whether ginsenoside-Rbl （Gs-Rb1） inhibits the apoptosis of hypoxia cardiomyocytes by up-regulating apelin-APJ system and whether the system is affected by hypoxia-induced factor 1α （Hif-1α）. Methods： Neonatal rat cardiomyocytes were randomly divided into 6 groups： a control group, a simple COCl2 group, a simple Gs-Rbl group, a CoCl2 and Gs-Rbl hypoxia group, a CoCl2 and 3-（5＇-hydroxymethyl-2＇-furyl）-1-benzylindazole （YC-1） group, a CoCl2 and YC-1 group and a Gs-Rbl group, in which YC-1 inhibits the synthesis and accelerates the degradation of Hif-la. The concentration of COCI2, Gs-Rbl and YC-1 was 500 μmol/L, 200 μ mol/L and 5 μmol/L, respectively; the apoptosis ratio was analyzed with a flow cytometer; and apelin, APJ and Hif-1 c= were assayed with immunocytochemistry, Western blot assays and reverse transcription polymerase chain reaction （RT-PCR）. Results：（1） The anti-apoptosis effect of Gs-Rbl on hypoxia cardiomyocytes was significantly inhibited by YC-1; （2） Hypoxia significantly up-graded the expression of mRNA and protein of apelin; this effect was further reinforced by Gs-Rbl and significantly inhibited by YC-1; （3） Gs-Rb1 further strengthened the expression of APJ mRNA and APJ proteins once hypoxia occurred, which was significantly inhibited by YC-1; （4） Gs-Rb1 significantly increased the expression of Hif-1α, which was completely abolished by YC-1; （5） There was a negative relationship between AR and apelin （or APJ, including mRNA and protein）, a positive correlation between apelin （or APJ） protein and Hif-1a protein, in hypoxia cardiomyocytes. Conclusion： The apelin-APJ system plays an important role in the anti-apoptosis effect of Gs- Rb1 on hypoxia neonatal cardiomyocytes, which was partly adjusted by Hif-1α.

关键词： ginsenosides-Rb1 cardiomyocytes hypoxia apelin APJ hypoxia-induced factor 1α

apelin inhibits motor neuron apoptosis in the anterior horn following acute spinal cord and sciatic nerve injuries

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Neural Regeneration Research 2011年第20期6卷 1525-1529页

作者： Zhiyue Li Weiguo Wang Qun Zhao Weiquan Ning Bin Yang Suling Zhang Siyin Feng Department of Orthopedics Third Xiangya Hospital Central South University Changsha 410013 Hunan Province China

Rat models of acute spinal cord injury and sciatic nerve injury were established. apelin expression in spinal cord tissue was determined. In normal rat spinal cords, apelin expression was visible; however, 2 hours post spinal cord injury, apelin expression peaked. apelin expression increased 1 day post ligation of the sciatic nerve compared with normal rat spinal cords, and peaked at 3 days. apelin expression was greater in the posterior horn compared with the anterior horn at each time point when compared with the normal group. The onset of neuronal apoptosis was significantly delayed following injection of apelin protein at the stump of the sciatic nerve, and the number of apoptotic cells after injury was reduced when compared with normal spinal cords. Our results indicate that apelin is expressed in the normal spinal cord and central nervous system after peripheral nerve injury. apelin protein can reduce motor neuron apoptosis in the spinal cord anterior horn and delay the onset of apoptosis.

关键词： apelin rat spinal Cord sciatic nerve injury cell apoptosis neural regeneration

High-throughput sequencing analysis of differentially expressed mi RNAs and target genes in ischemia/reperfusion injury and apelin-13 neuroprotection

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Neural Regeneration Research 2018年第2期13卷 265-271页

作者： Chun-mei Wang Xue-lu Yang Ming-hui Liu Bao-hua Cheng Jing Chen Bo Bai Neurobiology Institute Jining Medical University

miRNAs regulate a variety of biological processes through pairing-based regulation of gene expression at the 3＇ end of the noncoding region of the target miRNA, miRNAs were found to be abnormally expressed in ischemia/reperfusion injury models. High-throughput sequencing is a recently developed method for sequencing miRNAs and has been widely used in the analysis of miRNAs. In this study, ischemia/reperfusion injury models were intracerebroventricularly injected with 50 pg/kg apelin-13. High-throughput sequencing showed that 357 known miRNAs were differentially expressed among rat models, among which 78 changed to 〉 2-fold or 〈 0.5-fold. Quantita- tive real-time polymerase chain reaction was selected to confirm the expression levels of four miRNAs that were differentially expressed, the results of which were consistent with the results of high-throughput sequencing. Gene Ontology analysis revealed that the predicted targets of the different miRNAs are particularly associated with cellular process, metabolic process, single-organism process, cell, and binding. Kyoto Encyclopedia of Gene and Genome analysis showed that the target genes are involved in metabolic pathways, mitogen-ac- tivated protein kinase signaling pathway, calcium signaling pathway, and nuclear factor-KB signaling pathway. Our findings suggest that differentially expressed miRNAs and their target genes play an important role in ischemia/reperfusion injury and neuroprotection by apelin-13.

关键词： nerve regeneration RNA sequencing microRNA apelin ischemia/reperfusion injury neuroprotection neuropeptide high-throughput sequencing neural regeneration

Elevated high sensitive C-reactive protein and apelin levels after percutaneous coronary intervention and drug-eluting stent implantation

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2010年第8期11卷 548-552页

作者： Xin DU Jun-ping KANG Jia-hui WU Qiang LV Chao-shu TANG Chang-sheng MA Cardiovascular Division Beijing Anzhen Hospital Copital Medical University Beijing 100029 China Department of Physiology and Pathophysiology School of Basic Medical Sciences Peking University Beijing 100083 China

Objective:Percutaneous coronary intervention(PCI) triggers an acute inflammatory response,while sirolimus is known to have anti-inflammatory properties;the inflammatory system response to PCI after sirolimus-eluting stent placement remains *** purpose of this study is to determine the changes in high sensitive C-reactive protein(hs-CRP) and apelin after PCI procedure and drug-eluting stent implantation in patients with and without reduced left ventricular systolic ***:Forty-eight consecutive patients undergoing PCI at the Beijing Anzhen Hospital between July and September 2006 were ***-eluting stents were employed in all *** samples were drawn immediately before and 24 h after the *** hs-CRP and apelin levels were determined by enzyme ***:Paired t-test revealed a significant increase in both hs-CRP and apelin post-procedure(P=0.006 and Papelin levels compared to those with normal ventricular function [(46.8±10.8) vs.(72.0±8.4) pg/ml,Papelin increased to a level similar to the level of those with normal left ventricular systolic function 24 h after the PCI procedure [(86.7±11.6) vs.(85.1±6.1) pg/ml,P=0.72].Conclusions:hs-CRP and apelin levels increased after PCI and sirolimus-eluting stent *** with impaired left ventricular systolic function had significantly lower baseline apelin levels,which increased significantly after PCI.

关键词： High sensitive C-reactive protein (hs-CRP) apelin Percutaneous colonary intervention (PCl)

apelin： a double-edged sword in cardiac hypertrophy

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中国药理学通报 2015年第B11期31卷 161-162页

Aim As we all know, apelin acts as the endogenous ligand of APJ, being a member of G protein cou- pled receptors family, apelin/APJ system is involved in plentiful diseases and extremely responsible for the occur- rence and the development of cardiovascular diseases, among many kinds of heart diseases, it is the cardiac hyper- trophy that catches our attention. The myocardial expression of apelin/APJ decreased in rats with left ventrieular hypertrophy suggesting us there is a link between apelin and cardiac hypertrophy. Furthermore, it has been repor- ted that apelin is able to alleviate cardiac hypertrophy induced by Ang II, H202 and exercise. Nevertheless, our la- boratory discovered that apelin is certain to induce cardiac hypertrophy through PI3k-Akt-ERK1/2-p70S6K pathway or via up-regulating the levels of ROS to cause oxidative stress. The above-mentioned contradiction indicates us apelin may have dual effects in cardiac hypertrophy. Moreover, we also illuminate that apelin is involved in some diseases such as obesity, diabetes, hypertension, myoearditis and myocardial infarction, eoineidentally, all these diseases are associated with cardiac hypertrophy. Therefore, this review is aim to unveil the intricate relationship between apelin and cardiac hypertrophy.

关键词： apelin APJ Ang Ⅱ oxidative stress obesity hypertension myocardial infarction cardiac hypertrophy

apelin and vascular endothelial growth factor are associated with mobilization of endothelial progenitor cells after acute myocardial infarction

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The Journal of Biomedical Research 2012年第6期26卷 400-409页

作者： Jiaxin Ye Ping Ni Lina Kang Biao Xu Department of Cardiology Drum Tower Clinical College Affiliated with Nanjing Medical University Nanjing Jiangsu 210008 China Department of Cardiology Drum Tower Hospital (L.N.K.) Nanjing Jiangsu 210008 China

This study was designed to determine the levels of early endothelial progenitor cells （EPCs）, apelin, vascu- lar endothelial growth factor （VEGF） and stromal cell-derived growth factor-1 （SDF-1） after acute myocardial infarction （AMI）, and to investigate the relationships between these cytokines and early EPCs. Early EPCs, de- fined as CD133＋, KDR＋, and CD34~ cells, were quantified by flow cytometry. The levels of early EPCs and those cytokines in AMI patients were significantly different from those with coronary artery disease or controls （P 〈 0.05）. Plasma apelin levels were inversely correlated with Gensini score and early EPCs （both P 〈 0.01）. Early EPCs, VEGF and SDF-1 showed different patterns of changes in AMI patients during the first 24 h. The trend in the change of early EPCs was proportionally correlated with that of VEGF （P 〈 0.05）. AMI patients exhibited in- creased early EPCs with remarkably decreased apelin levels and enhanced VEGF levels.

关键词： apelin vascular endothelial growth factor (VEGF) stromal cell-derived growth factor-1 (SDF-1),endothelial progenitor cells (EPCs)

apelin/APJ:血管功能调节因子和药物治疗新靶点

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生物化学与生物物理进展 2015年第8期42卷 721-732页

作者：曹建刚李兰芳陈临溪南华大学药物药理研究所衡阳421001

apelin(APJendogenousligand)是血管紧张素Ⅱ1型受体相关蛋白(angiotensin receptor-like 1,APJ)的内源性配体.apelin/APJ系统在机体内广泛分布,在众多血管系统表达水平较高,如心血管系统、肺血管系统等.研究发现,apelin可调节血管张力... 详细信息

apelin(APJendogenousligand)是血管紧张素Ⅱ1型受体相关蛋白(angiotensin receptor-like 1,APJ)的内源性配体.apelin/APJ系统在机体内广泛分布,在众多血管系统表达水平较高,如心血管系统、肺血管系统等.研究发现,apelin可调节血管张力,促进血管平滑肌细胞增殖、视网膜血管新生以及单核细胞向内皮细胞黏附,促进肝门静脉和冠状动脉侧枝形成等.本文就apelin调节血管功能及其相关疾病(高血压、肺动脉高压、动脉粥样硬化、胶质瘤、肺癌、门静脉高压、糖尿病血管并发症等)进行综述,揭示了apelin与血管及其相关疾病的内在联系,表明apelin/APJ可作为血管疾病的治疗靶点.

关键词： apelin APJ 血管平滑肌细胞自噬血管新生血管重构高血压动脉粥样硬化

PI3K信号通路介导apelin-13促进单核细胞-血管内皮细胞黏附的研究(英文)

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生物化学与生物物理进展 2011年第12期38卷 1162-1170页

作者：毛小环苏桃张先慧李芳秦旭平廖端芳李兰芳陈临溪南华大学药物药理研究所湖南省高等学校药物蛋白质组学重点实验室湖南省动脉硬化学重点实验室衡阳421001 湖南省环境生物职业技术学院衡阳421001

本室以前已经报道G蛋白偶联受体APJ的内源性配体多肽apelin-13促进单核细胞-血管内皮细胞黏附,本文研究PI3K信号途径是否参与apelin-13促进单核细胞-血管内皮细胞黏附,探讨apelin/APJ系统的细胞信号转导机制.MPO方法检测细胞黏附;Wester... 详细信息

本室以前已经报道G蛋白偶联受体APJ的内源性配体多肽apelin-13促进单核细胞-血管内皮细胞黏附,本文研究PI3K信号途径是否参与apelin-13促进单核细胞-血管内皮细胞黏附,探讨apelin/APJ系统的细胞信号转导机制.MPO方法检测细胞黏附;Western blot方法检测PI3K、VCAM-1的表达.Western blot方法结果显示,apelin-13(0、0.5、1、2、4μmol/L)浓度依赖性刺激血管内皮细胞PI3K磷酸化,以1μmol/L最为明显;1μmol/L apelin-13时间依赖性促进血管内皮细胞PI3K磷酸化,在30 min增加最为显著;PI3K抑制剂LY294002明显抑制apelin-13诱导的VCAM-1表达和单核细胞-血管内皮细胞黏附.上述结果表明,PI3K信号途径介导apelin-13促进单核细胞-血管内皮细胞黏附.

关键词： apelin APJ PI3K 血管内皮细胞单核细胞

apelin protects against cardiomyocyte apoptosis induced by glucose deprivation

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Chinese Medical Journal 2009年第19期122卷 2360-2365页

作者： ZHANG Zhi YU Bo TAO Gui-zhou Department of Cardiology First Affiliated Hospital China Medical University Shenyang Liaoning 110001 China Department of Cardiology First Affiliated Hospital of Liaoning Medical University Jinzhou Liaoning 121001 China

Background Apoptosis is a major cause of ischemic heart dysfunction. apelin, the endogenous ligand for the G-protein-coupled APJ receptor, has been reported to exert cardioprotective effects during myocardial injury. The aim of this study was to investigate the effects of apelin on apoptosis of rat cardiomyocytes induced by glucose deprivation （GD） and study the related signaling pathway. Methods apelin and APJ mRNA expression were determined by RT-PCR in neonatal rat cardiomyocytes during different durations of GD. Cardiomyocyte apoptosis was detected by annexin V-FITC/propidium iodide （PI） staining after GD for 12 hours with or without apelin-13 （10 and 100 nmol/L） pretreatment. Protein levels of Akt and the mammalian target of rapamycin （mTOR） as well as cell apoptosis were detected in the presence or absence of LY294002 （a phosphatidylinositol 3-kinases （PI3K） inhibitor） or rapamycin （a mTOR inhibitor）. Results apelin mRNA expression was up-regulated when cardiomyocytes were exposed to GD for 6, 12, 18, and 24 hours compared with the base level （P 〉0.05, P 〈0.01, P 〈0.01, P 〈0.01）. However, when cardiomyocytes were exposed to GD for up to 36 hours, apelin mRNA expression was 17% lower than the base level （P〈0.05）. APJ mRNA expression paralleled that of apelin. apelin-13 pretreatment at 100 nmol/L significantly inhibited GD-induced cardiomyocyte apoptosis （P 〈0.05） and increased Akt and mTOR phosphorylation （P 〈0.01, P 〈0.01）. At the same time apelin-13 （100 nmol/L） up-regulated Bcl-2 protein expression and down-regulated Bax and cleaved caspase-3 expression （P 〈0.01, P 〈0.05, P 〈0.05）. The anti-apoptotic effect of apelin-13 was blocked by LY294002 （P 〈0.01） but not by rapamycin. Conclusions The endogenous apelin-APJ system is compensatorily up-regulated and ultimately down-regulated following sustained myocardial ischemia. apelin protects against ischemic cardiomyocyte apoptosis via activation of the PI3K/Akt pathway.

关键词： apelin apoptosis cardiomyocyte phosphatidylinositol 3-OH kinase (PI3K)/Akt