安苑铭；李妍静；张城林；丛馨；吴立玲；窦豆
AIM:To explore whether YAP protein is important in induced pluripotent stem cell ( iPSC)-induced cardiovascular progenitor cell and/or vascular smooth muscle differentiation .METHODS:Using episomal vector based reprogramming , we generated human iPSCs from donor fibroblasts .We used both this iPSCs and human H 1 embryonic stem cells to differentiate into vascular smooth muscle cells (VSMCs) through cardiovascular progenitor cells (CVPC).Western blotting, qPCR and immunofluorescence microscopy were used to check the expression of YAP and related genes during this differentiation process .RESULTS:The results showed that iPSCs expressed pluripotent stem cell markers, such as Oct4, Nanog, Sox2, TRA-1-60 and SSEA3, and could form teratoma in SCID mice.YAP was highly expressed in pluripotent stem cells , but dramatically decreased when CVPC differentiation started .YAP gradually increased dur-ing CVPC three-day differentiation.The TAZ and YAP binding partner TEAD1, but not TEAD2 and TEAD4, have similar expression pattern in CVPC differentiation .Immunofluorescence result confirmed that YAP was activated and accumulated in nucleus .Interesting-ly, both YAP and phosphorylated YAP expression decreased to very low level after CVPC differentiated into VSMCs in 7 days.TEAD4 and TAZ also decreased, while TEAD1, TEAD2 and TEAD3 expression did not change during VSMC differentiation .CONCLU-SION:YAP and TEAD1 expression increased during CVPC differentiation , while YAP and TEAD4 expression decreased from CVPC to VSMCs differentiation , which suggested YAP might have different function during diverse cell differentiation .
作者： 刊期： 2016年第08期
AIM:The direct renin inhibitor aliskiren displays antihypertensive and antialbuminuric effects in humans and in animal models . Emerging evidence has shown that aliskiren localizes and persists in medullary collecting ducts even after treatment was discontinued . The purpose of the present study was to investigate whether aliskiren regulates renal aquaporin expression and improves urinary concen -trating defect induced by lithium .METHODS:The mice were either fed with normal chow or LiCl diet (40 mmol/kg dry food per day for first 4 days and 20 mmol/kg dry food per day for last 3 days ) for seven days .Some mice were intraperitoneally injected aliskiren ( 50 mg/kg BW per day in saline ) .RESULTS:Mice injected aliskiren developed decreased urine output and increased urine osmolal -ity when compared with controls .Aliskiren significantly increased protein abundance of AQP 2 and phosphorylated-S256 AQP2 in the kidney inner medulla .Immunohistochemistry and immunofluoresence showed increased apical and intracellular labeling of AQP 2 and pS256-AQP2 in collecting duct principal cells of kidneys in mice treated with aliskiren .Aliskiren treatment prevented urinary concen-trating defect in lithium-treated mice , and improved the downregulation of AQP 2 and pS256-AQP2 protein abundance in inner medulla of the kidney .In primary cultured rat inner medulla collecting duct cells , aliskiren dramatically increased AQP 2 protein abundance which was significantly inhibited either by PKA inhibitor H 89 or by adenylyl cyclase inhibitor MDL 12330, indicating an involvement of the cAMP signalling pathway in mediating aliskiren-induced increased AQP 2 expression .CONCLUSION: The direct renin inhibitor aliskiren upregulates AQP 2 protein expression in inner medullary collecting duct principal cells and prevents lithium -induced nephro-genic diabetes insipidus ( NDI) likely via PKA-cAMP pathways .
作者： 刊期： 2016年第08期
目的：观察超声心动图斑点追踪方法能否早期发现和诊断异丙基肾上腺素（isoprenaline，ISO）引起的心脏功能异常。方法：将成年C57雄性小鼠分为对照组、ISO给药后3 d组和ISO给药后7 d组3组（ n＝6）。 ISO组均为一次性给予ISO 5 mg／kg皮下注射，对照组给予生理盐水皮下注射，分别于给药后3d和7d应用传统超声心动图方法以及斑点追踪方法评价小鼠心脏功能。结果：包括径向应变（radial strain，RS）、径向应变率（radial strain rate，RSR）和纵向应变（longitudinal strain，LS）在内的心脏应变分析指标，均在ISO注射后3 d开始显著降低。另外，与心肌梗死的局灶性改变不同，ISO诱导的心肌肥厚在应变分析中表现为全心功能的异常。而相比之下，传统超声心动图仅能在ISO注射后7 d检测出E／E’显著升高，提示心脏舒张功能异常，而反映收缩功能的左室短轴缩短率（ FS），以及反映心脏舒张功能的另外2个指标E／A和E’／A’均无显著差异。此外，心脏应变异常仅发生在ISO诱导的病理性心肌肥厚中，而并不出现在跑步训练诱导的生理性心肌肥厚中。结论：本研究发现基于超声心动图斑点追踪技术的心脏应变分析对心脏功能障碍的早期诊断比传统超声心动图更为敏感，而且可以用于区分病理性与生理性心肌肥厚。
作者：安祥博；张幼怡；宋峣 刊期： 2016年第08期
目的：随着饮食习惯和生活方式改变，非酒精性脂肪肝发病率近十年来明显增高。 FXR在胆汁酸代谢及糖脂代谢中起重要作用， FXR硫氢化修饰后对糖脂代谢影响机制尚不清楚。方法：本研究主要用modified biotin switch assay 检测FXR硫氢化修饰。分别用CSE腺病毒和siRNA过表达和敲低H2 S观察FXR变化。 Real－time PCR和Weastern blot等方法检测FXR及下游信号通路关键分子mRNA和蛋白水平变化。用高脂喂养小鼠给予H2 S供体，观察肝脏形态学变化。结果：内源性和外源性H2 S都可使FXR发生硫氢化修饰，并增强其转录活性，抑制糖脂代谢关键分子SREBP1－C表达，其下游FAS、ACC、PEPCK、G6Pase等脂肪酸从头合成基因和糖异生基因受到抑制，动物实验部分高脂饮食小鼠给予H2 S供体腹腔注射后，肝脏HE染色和油红O染色均表明脂滴变小，脂肪肝减轻。结论：研究提示FXR硫氢化修饰后抑制脂质合成，减轻脂肪肝。
作者：徐文静；范静慧；杜从阔；林宪娟；郑凤娇；蔡君艳；耿彬 刊期： 2016年第08期
目的：观察短暂反复缺血预处理（IPC）对肾缺血再灌注损伤（RI／RI）的影响。方法：成年健康雄性SD大鼠，体重150～180 g，随机分为：假手术（sham）组；缺血再灌（I／R）组；缺血预处理1次＋缺血再灌（1＋I／R）组；缺血预处理2次＋缺血再灌（2＋I／R）组；缺血预处理3次＋缺血再灌（3＋I／R）组。各组动物经1％戊巴比妥钠腹腔麻醉，开腹，暴露双侧肾动静脉。假手术组只开腹不夹闭双侧肾动静脉；I／R组夹闭双侧肾动静脉45 min后松夹再灌；1＋I／R、2＋I／R和3＋I／R各组则分别夹闭双侧肾动、静脉5 min后再灌5 min 1次、2次和3次后再行夹闭双侧肾动静脉45 min后松夹再灌；再灌后逐层缝合腹膜，腹壁肌肉和皮肤。24 h后取血及双侧肾脏测定血清肌酐（ SCr）、血尿素氮（ BUN）以及肾组织丙二醛（ MDA）、超氧化物歧化酶（SOD）的水平。结果：与I／R组相比，1＋I／R、2＋I／R和3＋I／R组BUN和SCr均明显降低（P＜0．05），其中以3＋I／R组BUN和SCr降低程度为著；I／R组MDA含量较sham组显著升高；SOD活力则较sham组明显降低（P＜0．01）；与I／R组相比，1＋I／R、2＋I／R和3＋I／R组SOD活力均明显增加，而MDA含量则明显降低（P＜0．05），其中以3＋I／R组MDA含量降低得为显著。结论：短暂反复IPC可改善RI／RI大鼠肾功能，减轻肾组织损伤。
作者：仝飞；牛丽静；王慧娟；苗智慧；夏晓红 刊期： 2016年第08期
AIM:To investigate regulatory roles of Apelin in adventitial remodeling and fibrosis in rats with transverse aortic constriction ( TAC) .METHODS:The male Sprague-Dawley rats with TAC were randomized to daily deliver either pyroglutamyl Apelin-13 ( 50μg/kg) or saline for 4 weeks.RESULTS:Histomorphometric analysis by HE and Masson Trichrome staining revealed increased medi -al and adventitial thicknesses , especially in the adventitia , in ascending aortas in rats with TAC when compared with the sham-operated rats.Downregulation of APJ receptor and elevations in phosphorylated mTOR and ERK 1/2 levels were observed in rats with TAC . There are marked increases in heart weight ( HW) , HW/body weight ratio , and aortic fibrosis in rats with TAC .The pressure over-load-mediated pathological adventitial remodeling was strikingly rescued by Apelin-13, associated with attenuation of aortic fibrosis and reduced mRNA expression of TGF-β1, fibronectin and collagen I .CONCLUSION:Our results demonstrate the importance of Apelin-13 in amelioration of aortic adventitial remodeling and fibrosis in rats with TAC via modulation of the mTOR /ERK signaling , thus indi-cating potential therapeutic strategies by enhancing Apelin /APJ action for preventing pressure overload-and fibrosis-associated cardio-vascular disorders .
作者： 刊期： 2016年第08期
目的：VEGF－VASH1（vasohibin－1）通路在血管新生过程的精细调节中发挥重要作用，转录因子锌指蛋白667（zinc finger protein 667， ZNF667）具有促进血管新生作用。本研究主要探讨ZNF667对上述通路中VEGF和VASH1表达的调控，以期阐明ZNF667促进血管新生的分子机制。方法：采用LAD建立慢性心肌缺血小鼠模型，采用HE染色和CD31免疫组化分析缺血心肌组织形态学变化及微血管形成。采用Matrigel、划痕和Transwell分析HUVEC管型形成和迁移；采用Western blot、ELISA和定量PCR分别检测ZNF667和VASH1蛋白质和mRNA表达；mRNA测序分析ZNF667过表达HUVEC的mRNA差异表达；染色质免疫沉淀（ chromatin immunoprecipitation ， ChIP）检测ZNF667与VEGF和VASH1启动子区结合情况。结果：免疫组化和HE染色显示与假手术组相比，缺血心肌中组织损伤加重伴随有CD31＋微血管数目增加，同时心肌组织中VEGF和ZNF667蛋白和mRNA表达呈时间依赖性增加，而VASH1表达降低。 mRNA测序、ELISA和定量PCR显示ZNF667过表达可促进HUVEC中VEGF表达而抑制VASH1表达。 VASH1过表达可抑制VEGF和ZNF667的促HUVEC管型形成和迁移作用。 ChIP显示ZNF667与VEGF（－346～－350 bp；－265～－269 bp）和VASH1（－170～－175 bp）基因启动子区结合。结论：HUVEC中ZNF667靶向调节VEGF－VASH1通路促进管型形成和迁移作用，这一调节机制可能参与缺血心肌中微血管新生过程。
作者：陈亦菲；邹江；王念；刘可；张华莉；王慷慨；肖献忠 刊期： 2016年第08期
目的：感染负荷被认为是动脉粥样硬化（ AS）新的独立危险因素。金黄色葡萄球菌（ S．aureus）是临床常见的致病菌之一。本课题组前期研究发现，金黄色葡萄球菌超抗原样蛋白－5（SSL5）可以激活血小板。我们推测，SSL5可能通过激活血小板而诱发炎症反应，探讨其机制可以为阐明感染负荷在AS中的作用提供新的实验证据。方法：体外培养人外周血单核细胞及THP－1细胞，以SSL5激活血小板所产生的微粒（ SSL5－PMPs）作用于上述细胞。结果：SSL5－PMPs呈时间和剂量依赖性地促进单核细胞IL－1β、TNF－α、MCP－1和MMP－9的表达；并促进MCP－1诱导的单核细胞迁移；阻断CD40L与CD40的相互作用，可以部分抑制SSL5－PMPs诱导单核细胞产生炎症介质；以siRNA下调单核细胞CD40或TRAF6基因的表达，导致SSL5－PMPs诱导单核细胞炎症介质的产生减少，并抑制NF－κB p65亚单位的磷酸化及核转位；阻断TLR4信号通路对SSL5－PMPs诱导单核细胞释放炎症介质没有影响。结论：SSL5可以激活血小板并产生PMPs；SSL5－PMPs与单核细胞结合，且主要与外周血中的具有促炎作用的单核细胞结合，促进炎性细胞因子的释放，CD40－TRAF6－NF－κB信号通路主要参与了这一过程。本研究为阐明感染负荷的致动脉粥样硬化机制提供了依据。
作者：胡厚源；贝俊杰；肇炜博 刊期： 2016年第08期
AIM:To investigate the regulation mechanism for insufficient KChIP 2 expression induces Ito,f downregulation and arrhythmogene-sis in cardiac hypertrophy .METHODS:Bidirectional manipulations of MG 53 expression were performed by adenoviral overexpression of MG53 or knockdown of MG53 with RNA interference in neonatal rat ventricular myocytes with or without PE stimulation .Ito,f was re-corded with patch clamp in whole-cell mode 48 h after adenoviral transfection .Then the WT or MG53 knockout ( MG53 -/-) mouse model of left ventricular hypertrophy induced by transverse aortic constriction ( TAC) were used to detect the susceptibility to ventricu-lar arrhythmia.RESULTS: Here, we show muscle-specific MG53 regulates KChIP2 expression and Ito,f densities, where they are downregulated in hearts from MG53 knockout mice and MG53 knockdown rat cardiomyocytes , but upregulated in MG53 overexpressed cells.MG53 expression is decreased in phenylephrine ( PE)-induced cardiomyocyte hypertrophy and restoration of MG 53 rescues PE-induced downregulation of KChIP2 and Ito,f.Furthermore, MG53 is decreased in a mouse model of hypertrophy induced by transverse aortic constriction and ablation of MG 53 increases the susceptibility to ventricular arrhythmia by exaggerating Ito,f remodeling.CON-CLUSION:These findings establish MG53 as a novel regulator of Ito,f and its central role in arrhythmogenesis in hypertrophy .
作者： 刊期： 2016年第08期
目的：葡萄糖转运体4（GLUT4）在球囊损伤血管新生内膜中高表达，而其转位过程依赖于肌动蛋白（actin）细胞骨架的调节。平滑肌蛋白22α（ smooth muscle protein 22α，SM22α）是一种actin细胞骨架相关蛋白，其在增殖性血管疾病中表达下调。本研究观察了SM22α是否参与血管损伤或者PDGF刺激诱导的GLUT4表达和转位活性升高。方法：用PDGF－BB刺激血管平滑肌细胞（ vascular smooth muscle cell ， VSMC），观察GLUT4膜转位和细胞骨架的变化；用荧光葡萄糖2－NBDG检测葡萄糖摄取；用特异性siRNA敲低内源性SM22α表达；BrdU实验检测细胞增殖；高效液相色谱法检测组织葡萄糖含量。结果：PDGF－BB诱导VSMCs GLUT4转位和葡萄糖摄取依赖于皮层F－actin聚合，而敲低SM22α促进这一过程。损伤新生内膜处GLUT4表达显著增加，PDGF－BB刺激促进细胞GLUT4表达和葡萄糖消耗，抑制GLUT4活性则显著降低细胞增殖活性。相对于WT组， SM22α－／－小鼠颈总动脉2－NBDG摄取显著增加，结扎后28 d新生内膜明显增厚，损伤动脉组织GLTU4转位和葡萄糖含量均明显升高。结论：PDGF－BB诱导的GLUT4转位和糖摄取参与VSMCs 增殖。缺失SM22α可诱导皮层细胞骨架聚合，增强PDGF－BB诱导的GLUT4膜转位和糖摄取及代谢活性。 SM22α是一种新的增殖相关糖代谢调节因子。
作者：赵丽丽；陈鹏；谢肖立；窦永青；聂磊；林燕玲；李晓坤；苗穗兵；董丽华；尹亚娟；张丹丹；宋昱；韩梅 刊期： 2016年第08期
目的：血管内皮损伤是糖尿病血管并发症的重要病理基础，早期研究发现Ca2＋稳态失衡参与内皮损伤，但是何种钙通道参与尚不十分明确。高糖可诱导内皮细胞上钙池操控性钙内流增加，同时内质网应激水平升高，因此本研究旨在研究Orai1通道对内质网应激反应的调控在糖尿病血管内皮损伤中的作用。方法：在动物及细胞水平利用Western blot 和real－time PCR检测糖尿病状态下Orai1通道的表达变化及内质网应激反应水平；进一步利用Orai1 shRNA腺病毒抑制该通道表达，在细胞水平观察其对高糖状态下内质网应激反应相关蛋白及内皮细胞磷酸化eNOS、NO生成的影响；在动物水平，利用血管张力测定仪检测Orai1通道对糖尿病小鼠血管内皮依赖性舒张反应的影响。结果：糖尿病状态下，Orai1表达及内质网应激水平均显著升高，抑制Orai1的表达可减少内质网应激标志物ATF4、CHOP、BiP等的表达，同时逆转内皮细胞NO生成水平及内皮依赖性血管舒张功能障碍。结论：Orai1通道参与糖尿病内皮损伤，其机制与调控内质网应激反应有关。
作者：杨慧；邝素娟；饶芳；薛玉梅；单志新；林秋雄；杨敏；吴书林；邓春玉 刊期： 2016年第08期
目的：探讨核仁素对心肌梗死后巨噬细胞极化的影响及其作用机制。方法：采用小鼠心肌梗死模型；采用心肌内注射核仁素RNA干扰的慢病毒载体，从整体水平观察核仁素低表达对小鼠心梗后死亡率的影响及对巨噬细胞极化的影响；采用流式细胞术检测巨噬细胞极化情况。结果：心肌梗死1 d后，核仁素表达减少，3 d后表达增加，5 d后明显升高，达（2．73±0．47）倍，7 d后有所下降。小鼠心肌梗死2 d、5 d后，心肌中巨噬细胞明显增多；心肌梗死2 d后心肌中M1型巨噬细胞占77．71％，而心肌梗死5 d后心肌中M2型巨噬细胞占82．13％。核仁素低表达可抑制心肌梗死5 d后M2型巨噬细胞的极化，但对巨噬细胞的侵润无明显影响，可明显减少心肌梗死后28 d的存活率。核仁素过表达可使巨噬细胞极化相关基因NOTCH1和STAT6的mRNA水平表达上调，而核仁素低表达可下调NOTCH1和STAT6的mRNA表达水平。结论：核仁素可调控心梗后巨噬细胞的极化，核仁素低表达可增加小鼠心梗后死亡率。
作者：蒋碧梅；吕青兰；李媛彬；刘梅冬；刘可；涂自智；肖献忠 刊期： 2016年第08期
AIM:MicroRNAs ( miRNAs) were recognized to play significant roles in cardiac hypertrophy .But, it remains unknown whether cyclin/Rb pathway is modulated by miRNAs during cardiac hypertrophy .This study investigates the potential roles of microRNA-1 (miR-1) and microRNA-16 (miR-16) in modulating cyclin/Rb pathway during cardiomyocyte hypertrophy .METHODS:An animal model of hypertrophy was established in a rat with abdominal aortic constriction (AAC).In addition, a cell model of hypertrophy was also achieved based on PE-promoted neonatal rat ventricular cardiomyocyte .RESULTS:miR-1 and-16 expression were markedly de-creased in hypertrophic myocardium and hypertrophic cardiomyocytes in rats .Overexpression of miR-1 and -16 suppressed rat cardiac hypertrophy and hypertrophic phenotype of cultured cardiomyocytes .Expression of cyclins D1, D2 and E1, CDK6 and phosphorylated pRb was increased in hypertrophic myocardium and hypertrophic cardiomyocytes , but could be reversed by enforced expression of miR-1 and -16.CDK6 was validated to be modulated post-transcriptionally by miR-1, and cyclins D1, D2 and E1 were further validated to be modulated post-transcriptionally by miR-16.CONCLUSION: Attenuations of miR-1 and -16 provoke cardiomyocyte hypertrophy via derepressing the cyclins D1, D2, E1 and CDK6, and activating cyclin/Rb pathway.
作者： 刊期： 2016年第08期
目的：探讨棕榈酸激活上皮细胞钠通道（ epithelial sodium channel ， ENaC）的分子机制，以及H2 S对抗棕榈酸引起的ENaC异常激活的作用和机制。方法：应用肾皮质集合管上皮细胞，采用膜片钳技术研究H2 S对抗棕榈酸引起ENaC异常激活的保护作用和分子机制；应用激光共聚焦显微镜技术观察棕榈酸能否调节细胞内钙水平和细胞内ROS水平变化。结果：棕榈酸引起的细胞ENaC活性升高可以被NaHS抑制；棕榈酸引起的细胞内ROS水平升高可以被NaHS抑制，且应用NADPH抑制剂APO可以抑制棕榈酸引起的ENaC活性升高；棕榈酸可以引起细胞内钙的升高；应用钙离子螯合剂BAPTA／AM或IP3受体抑制剂APB可以抑制棕榈酸引起的ENaC活性升高；胰岛素受体抑制剂HNMPA和PI3K抑制剂LY204002也可以抑制棕榈酸引起的ENaC活性升高；DTT可以模拟NaHS对棕榈酸引起ENaC异常激活的保护作用。结论：棕榈酸通过诱导胰岛素受体磷酸化，引起细胞内钙释放，进而激活NADPH升高细胞内活性氧水平，引起ENaC异常激活。气体信号分子H2 S通过氧化还原反应抑制棕榈酸引起的ENaC异常激活。
作者：王秋石；梁辰 刊期： 2016年第08期
目的：动物实验表明高盐摄入可降低循环及肾脏中肾胺酶的表达水平。本研究拟探讨钠、钾摄入对成人血清和尿中肾胺酶表达的影响。方法：42名（28～65岁）来自中国北方农村的受试者参与了这项研究。所有受试者依次接受低盐饮食7 d（氯化钠3 g／d），高盐饮食7 d（氯化钠18 g／d），高盐补钾饮食7 d（氯化钠18 g＋氯化钾4．5 g／d）。血清及尿中肾胺酶水平用ELISA试剂盒进行检测。结果：低盐饮食期，血清中肾胺酶水平较基线期显著升高。低盐转向高盐饮食期时，血清肾胺酶水平随之下降，但同时给予补钾后，可阻止高盐所致的肾胺酶水平下降。尿中肾胺酶水平在高盐饮食期显著高于低盐期。高盐补钾期，尿中肾胺酶水平与单纯高盐期相比无显著差异，但显著高于低盐饮食期。24 h尿钠排泄与血清中肾胺酶水平呈负相关，与尿中肾胺酶水平呈正相关。结论：饮食中钠、钾含量的变化可显著影响中国人血清及尿中肾胺酶的表达水平。
作者：吕永波；汪洋；牟建军 刊期： 2016年第08期
目的：探究心脏心肌细胞与成纤维细胞间通过远距离连接———膜纳米管直接传输线粒体的生理学意义以及具体的传输机制。方法：通过激光共聚焦仪器的活细胞追踪功能，实时观测线粒体的运动及传输；利用免疫荧光双染的方法，观察线粒体与微管、微丝等结构的共定位，并使用微管抑制剂Nocodazole ，阻断微管的存在，以观察微管对线粒体传输的作用；使用West－ern blot、real－time PCR方法检测KIF5B蛋白的表达，继而通过使用转染KIF－5B－siRNA慢病毒方法敲减细胞中KIF5B，探究KIF5B的作用；运用TUNEL染色及高内涵检测的方法，检测心肌细胞的凋亡。结果：原代乳大鼠心肌细胞与成纤维细胞间形成的膜纳米管可以传输线粒体，线粒体在其中的平均运动速度（17．5±2．1） nm／s。在膜纳米管中，线粒体与微管存在共定位，线粒体的传输依赖于微管，并且KIF5B是膜纳米管中推动线粒体传输的马达蛋白，使线粒体可以从成纤维细胞中传输到心肌细胞中。在心肌细胞的缺氧－复氧模型的病理模型中，成纤维细胞通过将自身的线粒体传输到心肌细胞中，起到减少心肌细胞凋亡的保护作用。结论：作为一种新型的细胞连接，心肌细胞与成纤维细胞间形成的膜纳米管可以通过直接传输线粒体，从而减少受损心肌细胞的凋亡，这种传输依赖于微管及马达蛋白KIF5B。
作者：张江晖；沈静；吴济民；肖晗；何康敏；吕志珍；李子健；徐明；张幼怡 刊期： 2016年第08期
AIM:To analyze the proteins included in exosomes derived from blood of patients with hypertension and seek the main pathologi -cal changes in hypertension .METHODS:Forty-seven patients and healthy subjects were recruited and divided into two comparisons :healthy subjects vs atherosclerosis ( HS vs AS) , and atherosclerosis vs hypertension plus atherosclerosis ( AS vs HT+AS) .We extrac-ted exosomes from blood and utilized LC-MS/MS to identify the protein expression .We used GO analysis to established the hierarchy programs of biological process and molecular function .PPI was used to find the proteins related to the terms .RESULTS:It was found that three final child terms repeatedly shown in BP of the two categories ( HS vs AS and AS vs HT+AS):“signal transduction in re-sponse to DNA damage”,“response to zinc ion”, and“platelet aggregation”.It was found that two final child terms in MF of the two categories:“interleukin 2 receptor binding” and“ploy(A) RNA binding”.The proteins, PSMA6, PSMA7 and CA2, were related to the terms in the two categories .CONCLUSION: We discovered that the exosome proteins may indicate the pathological changes in hypertension through the biological processes related with the specific proteins .These specific proteins, such as VCL, PSMA6, DP, AKAP, ATP5B and CA2, can be the new indicators for severity of hypertension and new therapeutic targets .
作者： 刊期： 2016年第08期
目的：IL－37是白细胞介素－1家族成员，可调节血管生成，抑制肿瘤生长，对I／R损伤、炎症性肠道疾病和类风湿性关节炎等有保护作用。由于炎症反应能导致组织氧供应失衡，使局部组织细胞处在缺氧的微环境，因而，本实验主要针对IL－37是否在缺氧环境中发挥细胞保护作用进行研究。方法：分别给予1％和21％ O2培养上皮细胞。通过瞬时转染使细胞过表达IL－37，24 h后提取RNA和蛋白质。利用RT－PCR检测IL－37、TNF－α、IL－1、IL－6和CXCL2 mRNA的表达；利用Western blot 检测PARP的总蛋白和剪切蛋白的表达量。利用流式细胞术检测细胞凋亡。结果：缺氧时，细胞凋亡增加1．63倍，内源性IL－37和促炎因子mRNA水平的表达均显著增加，PARP总蛋白表达增加2．1倍。 IL－37可显著抑制缺氧诱导的细胞凋亡，并显著降低IL－6和CXCL2的mRNA水平，但其对可以修复DNA的PARP总蛋白的表达在缺氧时无明显作用， caspase－3剪切的PARP也无显著差异。结论：在缺氧环境中，IL－37作为固有免疫调节因子，可以通过抑制细胞凋亡，炎性因子IL－6和CXCL2的表达抑制局部的过度炎症反应。但TNF－αmRNA的表达和caspase－3剪切底物PARP的剪切蛋白无显著变化，提示IL－37不是通过减少TNF－α的表达、进而减少FADD和caspase－8的募集、再减少活化caspase－3的途径抑制细胞凋亡的。其具体抑制凋亡的机制有待进一步研究。
作者：周梦晨；姜桂青；李倩倩；王孟茹；廖玉华；凃欣 刊期： 2016年第08期
AIM:To investigate the effects of human induced pluripotent stem cell-derived exosomes (hiPSC-exo) on cell viability, capillary-like structure formation , and senescence in endothelial cells exposed to high glucose .METHODS: Exosomes were isolated from the conditional medium of hiPSCs and confirmed by transmission electron microscopy , nanoparticle tracking analysis , and Western blot analysis using Alix and CD63 as markers.hiPSC-exo were labeled with PKH26 for tracking.Cultured HUVECs were treated with high glucose (33 mmol/L) with or without hiPSC-exo (20 mg/L) for 48 h, and cell viability, capillary tube formation, and senescence were assessed .RESULTS:hiPSC-exo showed a typical cup shape and could be taken up by human umbilical vascular endothelial cells (HUVECs) in a concentration-dependent manner.When exposed to high glucose, viability and tube formation in HUVECs was signifi-cantly reduced, whereas the proportion of senescent cells was higher compared to that in control HUVECs (P<0.01).Furthermore, hiPSC-exo restored cell viability and capillary-like structure formation , and reduced senescence in HUVECs exposed to high glucose (P<0.01).However, hiPSC-exo had minimal effects on normal HUVECs.Therefore, stem cell-derived exosomes can promote cell proliferation, enhance capillary-like structure formation , and reduce senescence in endothelial cells exposed to high glucose . CONCLUSION:Our study highlights the role of exosomes derived from hiPSC and may provide a new strategy for maintaining vascular health, preventing vascular aging , and avoiding pathological vascular remodeling that occurs in many diseases .
作者： 刊期： 2016年第08期
目的：C1q／肿瘤坏死因子相关蛋白－3（CTRP3）是一种脂肪细胞因子，它与多种代谢性及心血管疾病密切相关，但CTRP3对线粒体生物生成的影响尚不清楚。本研究主要探讨CTRP3对心肌细胞线粒体生物生成的影响及相关机制。方法：原代培养乳大鼠心肌细胞并给予CTRP3处理。使用PCR、Western blot和免疫共沉淀等方法分别检测线粒体生物生成相关蛋白、线粒体DNA拷贝数、ATP含量和sirtuin 1（ SIRT1）活性的变化。结果：CTRP3显著增加过氧化物酶体增殖激活受体共激活因子1α（ PGC－1α）、核呼吸因子1（ NRF－1）、线粒体转录因子A（ TFAM）、线粒体氧化磷酸化复合物III和V的表达。 CTRP3显著升高心肌线粒体DNA拷贝数和ATP含量，而在心肌细胞中敲低PGC－1α可使上述效应减弱。预孵育腺苷酸活化蛋白激酶（ AMPK）的抑制剂AraA可以逆转由CTRP3引起的NRF－1、TFAM和复合物III、V的表达升高。 CTRP3可上调SIRT1的表达和活性，SIRT1抑制剂EX－527可阻断CTRP3对PGC－1α的去乙酰化调节作用。此外，CTRP3对SIRT1表达和活性的促进作用也可被AraA所阻断。结论：CTRP3通过AMPK／PGC－1α通路促进心肌细胞线粒体生物生成。
作者：张城林；冯寒；李丽；王瑾瑜；张艳；吴立玲 刊期： 2016年第08期
中国病理生理杂志
主管:中国科学技术协会
主办：中国病理生理学会
