特异性敲除Apelin基因对单侧输尿管梗阻小鼠肾间质纤维化的影响

目的观察多肽Apelin基因缺失对单侧输尿管梗阻(UUO)小鼠肾间质纤维化的影响,探讨在梗阻性肾病肾纤维化形成中Apelin的作用和可能机制。方法利用CRISPR/Cas9技术构建Apelin基因敲除小鼠(KO)共20只,同时选取野生型小鼠(WT)20只,均为8周龄雄性C57BL/6J种系,采用随机数字表法分为:KO+Sham组、KO+UUO组、WT+Sham组、WT+UUO组,每组各10只。分别对小鼠行单侧输尿管结扎(UUO)或假手术(Sham)。术后2周将小鼠处死,取血浆和手术侧肾脏进行检测。采用Masson染色观察肾脏纤维化程度,免疫组织化学染色检测肾脏基质成分纤维连接蛋白(FN)、Ⅰ型胶原(Col-I)的分布和表达情况。采用Western blotting法检测肾小管上皮标志物E-钙黏素(E-cadherin)、间充质标志物α-平滑肌肌动蛋白(α-SMA),以及致纤维化细胞因子--转化生长因子-β1(TGF-β1)及其受体TGFβ-R1的蛋白表达量。采用酶联免疫吸附法(ELISA)检测小鼠血浆肌酐(Cr)、尿素氮(BUN)水平。结果KO+UUO组小鼠肾脏可见明显的肾小管萎缩和肾间质纤维化,基质成分FN和Col-I表达量显著升高。WT+UUO组小鼠肾脏的纤维化程度和基质的表达量均低于KO+UUO组,差异具有统计学意义(P<0.05)。KO+Sham组和WT+Sham组肾脏无明显的纤维化改变和基质的沉积。KO+UUO组小鼠,可见显著的肾小管上皮-间充质转分化(EMT)表现,E-cadherin减少,α-SMA增多,TGF-β1和TGFβ-R1受体的表达量也明显升高。与KO+UUO组相比,WT+UUO组肾小管上皮E-cadherin的表达较多,α-SMA、TGF-β1和TGFβ-R1受体的表达均较低,差异具有统计学意义(P<0.05)。而WT+Sham组和KO+Sham组小鼠上述标志物的表达趋于正常水平。KO+UUO组小鼠血浆Cr、BUN的水平最高,WT+UUO组小鼠上述标志物的水平均低于KO+UUO组,差异具有统计学意义(P<0.05)。KO+Sham组小鼠血浆Cr、BUN与WT+Sham组相比,差异无统计学意义(P>0.05)。结论Apelin在正常状态下对肾功能无明显影响,但在输尿管梗阻状态下其基因缺失可能是引起肾间质纤维化加重的因素。     

ELA-Apelin-APJ系统在血管重构稳态与血管疾病中的调控作用及相关药物研发进展

Apelin受体（Apelin receptor,APJ）是一种G蛋白偶联受体,其内源性配体包括Elabela （ELA）和Apelin,在血管重构稳态和损伤修复中发挥重要作用。ELA-Apelin-APJ系统参与血管的生成、分化及纤维化,以及细胞凋亡、炎症反应、氧化应激及血压调控,其活性和表达异常参与动脉粥样硬化、高血压、肺动脉高压、脑卒中等血管疾病的发病过程。概述ELA-Apelin-APJ系统在血管重构稳态中的调控作用及其作为潜在药物靶点的研究新进展,以期为血管研究提供参考。     

合并高血压的冠心病患者术前血清Apelin浓度与非体外循环冠状动脉旁路移植术后新发房颤的相关性研究*

【】 目的：研究合并高血压的冠心病患者术前血清Apelin检测值与非体外循环冠状动脉旁路移植(off-pump coronary artery bypass grafting , OPCABG)术后新发房颤(post operation atrial fibrillation, PoAF)的相关性,探讨血清Apelin值对PoAF的预测作用。方法：回顾性分析我院2014年10月1日至12月31日间的OPCABG手术病例,仅纳入合并高血压的冠心病患者共154例,收集围术期资料。使用统计学方法分析导致OPCABG-PoAF的相关危险因素,进行单因素和多因素logistic回归分析筛选独立危险因素。结果：吸烟、总胆固醇、左室舒张末期容积和术前血清Apelin值与OPCABG- PoAF具有相关性,其中总胆固醇(B=0.874,Wald =8.506,P=0.004,OR=2.396 、95%CI：1.332-4.310)、左室舒张末期容积(B=0.017,Wald =6.305,P=0.012,OR=1.018 、95%CI：1.004-1.031)和术前血清Apelin值(B=0.000,Wald =4.107,P=0.043,OR=1.000 、95%CI：0.999-1.000)分别是是OPCABG- PoAF的独立危险因素。结论：血清Apelin值降低是OPCABG- PoAF的独立危险因素,术前血清Apelin值检测对OPCABG- PoAF具有预测作用。     

Apelin triggers macrophage polarization to M2 type in head and neck cancer

Cancer comes after cardiovascular diseases in terms of mortality rate in the world. Chemotherapy, radiotherapy and surgical interventions are the current cancer treatment. Recently, it has been observed that immunotherapeutic approaches provide a significant improvement when used along with these interventions. The mononuclear system mainly consists of macrophages that play an active role in the pathology of many diseases because of having high plasticity capacities. Previous research suggested that they can be used as an alternative to cancer treatment. Aim was to investigate the effect of apelin on macrophage polarization in the tumor microenvironment.Mouse macrophage cell line RAW264.7 cells and head and were chosen for this study. The apelin expression was knockdown in neck cell carcinoma cell line SCCL MT1 cells using shRNA technique. SCCL MT1 cells having normal or suppressed apelin expression were co-cultured with mouse macrophage RAW264.7 cells. The effect of co-culturing on the expression of inflammatory genes in RAW264.7 cells was investigated.Suppressed apelin expression in SCCL MT1 cells resulted in elevated pro-inflammatory response in co-cultured macrophages. Expression of the IL1β, IL6, and TNFα genes significantly increased, however anti-inflammatory cytokine levels were significantly decreased. However, in the control group, a downregulation was determined in pro-inflammatory genes, while an increase was observed in anti-inflammatory genes. The protein levels of these cytokines in concordance with the RT-PCR analysis.As a result of this study, apelin released from cancer cells was found to affect macrophage polarization. These results indicated that the apelin peptide may cause the intense presence of M2-type macrophages in the tumor niche, and the therapeutic approaches targeting of apelin in cancer cells may have a potential role in macrophage polarization.