董思敏；王海璐；王全凯；张晶
目的：探究张应变条件下microRNA－33（miR－33）调控移植静脉内膜增生的机制，为缓解静脉移植内膜增生提供潜在治疗方法。方法：SD大鼠进行“套管法”自体静脉移植，Elastin－van Gesion染色观察内膜增生情况。使用FX4000细胞应力加载装置（ Flexcell International ）对静脉平滑肌细胞加载频率1．25 Hz、幅度10％的张应变以模拟静脉在动脉环境受到的张应变力学刺激。 qRT－PCR检测miR－33表达，Western blotting检测相关蛋白，BrdU增殖实验和CCK－8试剂盒检测细胞增殖。双萤光素酶报告基因验证miR－33与靶基因的作用关系。骨形态发生蛋白3（BMP3）特异性siRNA干扰片段、重组蛋白以及miR－33 in－hibitor和mimics用于研究细胞功能和相关信号通路。在体局部注射miR－33 agomir和antagomir来验证miR－33在静脉移植内膜增生中的作用。结果：移植静脉出现明显内膜增生，miR－33显著降低，而BMP3、p－Smad5和p－Smad2表达明显上升；牵拉条件下得到与移植静脉中相同的结果。双萤光素酶报告基因实验证明BMP3是miR－33的靶基因。 miR－33 mimics抑制BMP3及下游信号分子p－Smad2、p－Smad5表达和细胞增殖； miR－33 inhibitor 或者BMP3重组蛋白得到类似结果。在体注射miR－33 agomir降低BMP3及下游信号分子表达，亦可缓解静脉移植内膜增生。结论：miR－33－BMP3－Smad信号通路参与移植静脉平滑肌细胞增殖；miR－33可以缓解静脉移植内膜增生过程，具有潜在临床应用前景。
作者：黄凯；包晗；严志强；王璐；张萍；姚庆苹；施茜；陈小虎；王凯旋；沈宝荣；齐颖新；姜宗来 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
线粒体作为细胞能量代谢的重要场所，通过氧化磷酸化过程生成ATP为细胞供能。近年的研究表明，线粒体除具有能量生成功能之外，还参与母性遗传、多种生物大分子代谢以及细胞程序性死亡等病理过程。由此可见，线粒体的自身内源性平衡直接决定细胞的命运，线粒体的生成和自噬保持动态平衡。神经元主要以氧化磷酸化提供能量，维持神经元内各种生物学功能的完整，因此神经元内线粒体总体积分数约占细胞体积的30％。脑组织缺血再灌注将导致神经元线粒体结构破坏，线粒体DNA数量减少，生成能力降低，终引起线粒体氧化磷酸化功能下降，导致神经元能量匮乏而凋亡。
作者：王来；祝世功 刊期： 2016年第08期
AIM:To investigate whether KCNE 2 participates in the development of pathological hypertrophy .METHODS:Bidirectional ma-nipulations of KCNE2 expression were performed by adenoviral overexpression of KCNE 2 or knockdown of KCNE2 with RNA interfer-ence in PE-induced neonatal rat ventricular myocytes .Then overexpression of KCNE 2 in mouse model of left ventricular hypertrophy in-duced by transverse aortic constriction (TAC) by ultrasound microbubble-mediated gene transfer were used to detect the therapeutic function of KCNE2 in the development of hypertrophy .RESULTS:KCNE2 expression was significantly decreased in PE-induced hy-pertrophic cardiomyocytes and in hypertrophic hearts produced by TAC .Knockdown of KCNE2 in cardiomyocytes reproduced hypertro-phy, whereas overexpression of KCNE2 attenuated PE-induced cardiomyocyte hypertrophy .Knockdown of KCNE2 increased calcineurin activity and nuclear NFAT protein level , and pretreatment with nifedipine or FK 506 attenuated decreased KCNE 2-induced cardiomyo-cyte hypertrophy .Overexpression of KCNE 2 in heart by ultrasound microbubble-mediated gene transfer suppressed the development of hypertrophy and activation of calcineurin-NFAT and MAPK pathways in TAC mice .CONCLUSION:These findings demonstrate that cardiac KCNE2 expression is decreased and contributes to the development of hypertrophy via activation of calcineurin -NFAT andMAPK pathways .
作者： 刊期： 2016年第08期
Angiotensin-converting enzyme 2 (ACE2)-angiotensin (1-7) [Ang (1-7)]-Mas constitutes the vasoprotective axis and is demon-strated to antagonize the vascular pathophysiological effects of the classical renin -angiotensin system .We hypothesize that upregulation of ACE2-Ang (1-7) signaling protects endothelial function through reducing oxidative stress , thus resulting in beneficial outcome in di-abetes.Ex vivo treatment with Ang (1-7) augmented endothelium-dependent relaxation (EDR) in renal arteries from diabetic patients . Both Ang (1-7) infusion via osmotic pump (500 ng? kg -1? min-1 ) for 2 weeks and exogenous ACE 2 overexpression mediated by ad-enoviral ACE2 via tail vein injection rescued the impaired EDR and flow-mediated dilatation ( FMD) in db/db mice.Diminazene acetu-rate treatment (15 mg? kg-1? d-1 ) activated ACE2, increased the circulating Ang (1-7) level, and augmented EDR and FMD in db/db mouse arteries.In addition, activation of the ACE2-Ang (1-7) axis reduced reactive oxygen species (ROS) overproduction de-termined by dihydroethidium staining , CM-H2DCFDA fluorescence imaging , and chemiluminescence assay in db/db mouse aortas and also in high-glucose-treated endothelial cells .Pharmacological benefits of ACE 2-Ang ( 1-7 ) upregulation on endothelial function were confirmed in ACE2 knockout mice both ex vivo and in vitro.We elucidate that the ACE2-Ang (1-7)-Mas axis serves as an important signal pathway in endothelial cell protection in diabetic mice , especially in diabetic human arteries .In summary, endogenous ACE2-Ang (1-7) activation or ACE2 overexpression preserves endothelial function in diabetic mice through increasing nitric oxide bioavail -ability and inhibiting oxidative stress , suggesting the therapeutic potential of ACE 2-Ang(1-7) axis activation against diabetic vasculop-athy.
作者： 刊期： 2016年第08期
目的：探讨omega－3多不饱和脂肪酸（ polyunsaturated fatty acid ， PUFA）对动脉粥样硬化斑块稳定性的作用及可能机制。方法：采用低密度脂蛋白受体敲除（Ldlr－／－）小鼠喂养西方饮食（western diet， WD）6周诱导动脉粥样硬化，并在饮食中添加或不添加3％omega－3 PUFA进行干预。使用液相质谱联用检测血浆中PUFA及其代谢产物浓度。油红O染色分析测定动脉树斑块面积及主动脉根部斑块脂质含量，HE染色分析主动脉根部斑块大小，天狼星红染色分析胶原纤维含量，免疫荧光检测巨噬细胞和平滑肌细胞含量。结果：（1）与WD组相比，omega－3组小鼠动脉树斑块面积比例显著降低。（2） Omega－3处理组与对照组相比，主动脉根部斑块面积、脂质含量和巨噬细胞含量显著降低；同时胶原纤维和平滑肌细胞含量显著上升，斑块不稳定指数下降（P＜0．05）。（3）Omega－3处理组血清中omega－3 PUFA显著增加，分析代谢产物发现EEQ和18－HEPE水平显著增加。结论：Omega－3处理减少动脉粥样硬化斑块面积、增加斑块稳定性，其机制可能与其代谢产物水平变化相关。
作者：刘亚晋；李丹；张栩；艾玎；朱毅 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
AIM:Programmed necrosis ( necroptosis ) and apoptosis are crucially involved in multiple severe cardiac pathological conditions , including myocardial infarction, ischemia/reperfusion (I/R) injury, and heart failure.Whereas apoptotic signaling is well defined, the mechanisms underlying cardiomyocyte necroptosis remain elusive .METHODS AND RESULTS:Here we show that both mRNA and protein levels of receptor-interacting protein 3 (RIP3) in the hearts are increased by I/R injury and doxorubicin (Dox) treatment. In mice, RIP3 deficiency ameliorates myocardial necroptosis and heart failure induced by I /R (30-min ischemia/4-h or 8-week reper-fusion) or Dox treatment (20 mg/kg or 5 mg/kg ×4, i.p.).RIP3 overexpression induces cardiomyocyte necroptosis evidenced by de-creased intracellular ATP level and increased lactate dehydrogenase concentration in cell culture medium .RIP3 triggers myocardial ne-croptosis via activation of Ca2+/calmodulin-dependent protein kinase II (CaMKII), rather than the well-established RIP3 partners, RIP1 and MLKL (mixed lineage kinase domain-like protein).Specifically, our data indicate that I/R and Dox markedly activate myo-cardial CaMKII in wild-type but not RIP3-deficient mice , and that CaMKII inhibition or RIP 3 deficiency protect the heart from I/R-and Dox-induced cardiomyocyte necroptosis , cardiac remodeling and heart failure .Mechanistically , RIP3 activates CaMKII via both di-rect phosphorylation and indirect reactive oxidative species-dependent oxidation , and subsequently triggers opening of the mitochondrial permeability transition pore ( mPTP) and myocardial necroptosis .CONCLUSION: These findings identify CaMKII as a novel RIP 3 substrate and delineate a RIP3-CaMKII-mPTP myocardial necroptosis pathway , a promising target for the treatment of cardiac ischemic and oxidative damage , and heart failure .
作者： 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：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
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
目的：研究全反式维甲酸（ all－trans retinoic acid，ATRA）对胃癌细胞SGC－7901存活率与放射敏感性的影响，并讨论其可能的机制。方法：MTT法检测细胞存活率；平板克隆形成实验和流式细胞术分别检测细胞的放射敏感性和细胞周期；实时荧光定量PCR（ RT－qPCR）检测细胞中Bax、Bcl－2、survivin与NF－κB的mRNA表达。结果：ATRA可降低SGC－7901细胞存活率，当浓度到达8μmol／L时，抑制作用达到大；ATRA联合X射线处理后，与单纯放射处理相比，平均致死剂量（D0）和准阈剂量（Dq）显著变小（P＜0．05），且拟合的生存曲线明显下移；ATRA能显著降低放射诱导的细胞G2／M期阻滞，下调SGC－7901细胞Bcl－2与survivin的mRNA表达（ P＜0．05），上调Bax与NF－κB的mRNA表达（P＜0．05）。结论：ATRA能够增加胃癌细胞SGC－7901的凋亡及放射敏感性，可能与抑制细胞周期G2／M期的阻滞作用、下调Bcl－2与survivin mRNA表达和上调NF－κB与Bax mRNA表达有关。
作者：王艳萍；赵先群；张向东；许威；向晓辉 刊期： 2016年第08期
AIM:There is little evidence proving the molecular mechanism of WenxinKeli ( WXKL) .This study tried to explore the gene ex-pression profile and pathology alteration of WXKL-treated rabbits with myocardial infarction .METHOD: Twenty male adult rabbits were randomly divided into 4 groups:sham, model, WXKL and captopril groups .Model, WXKL and captopril groups underwent the ligation of the left anterior descending coronary artery , while sham group went through an identical procedure without ligation .WXKL (817 mg? kg-1? d-1), captopril (8 mg? kg -1? d-1) and distilled water (model and sham) were administered orally to the rabbits. 4 weeks later, hearts were taken out for expression chip and pathological staining (HE, Masson and TUNEL) after echocardiography. RESULT:WXKL could down-regulate genes associated with inflammation (CX3CR1, MRC1, and FPR1), apoptosis (cathepsin C and TTC5) and neuro-hormonal system (ACE and EDN1), and up-regulate angiogenesis promoting gene like RSPO 3, which explained why WXKL group represented with better cardiac function , less histopathological injury and slighter apoptosis .CONCLUSION:WXKL plays an important role in suppressing inflammation , inhibiting renin-angiotensin system and alleviating apoptosis , and might be a promising Chinese medicine in treating patients with myocardial infarction .
作者： 刊期： 2016年第08期
AIM:Increasing evidence suggests that carbohydrate-binding proteins play an essential role in tumor growth and metastasis .Ga-lectin-3, a multifunctional protein of an expanding family of β-galactoside-binding animal lectins , is the major nonintegrin cellular laminin-binding protein , and is implicated in a variety of biologic events , such as inflammation and angiogenesis .Because galectin-3 expression was shown to participate in mediating tumor angiogenesis and initiate signaling cascades in several diseases .We hypothe-sized that galectin-3 may promote pulmonary vascular endothelial neovascularization .METHODS:Hypoxic and MCT rat model of pul-monary artery remodeling was used .The mRNA and protein levels of galectin-3 in rats were measured by in situ hybrization and West-ern blot analysis.Endothelial cell (EC) proliferation, migration and tube formation were measured using MTT , cell scratch and Matri-gel assays, respectively.Protein expression was quantitated by Western blot analysis .LC 3A/B staining was detected with cellular im-munofluorescence staining .RESULTS:We found that galectin-3 was localized on the intima and adventitial wall .Galectin-3 was in-creased after rat hypoxia and MCT administration .Galectin-3 promoted EC proliferation , migration and tube formation , while its roles were reversed by RNA interference.Galectin-3 induced Atg 5, Beclin-1, LAMP-2, and LC 3A/B expression increases.Galectin-3 al-so increased LC 3A/B staining in ECs.Akt/mTOR and GSK-3βsignaling pathways were activated after galectin-3 treated ECs using its specific phosphorylation antibodies , while blocked it with LY294002 inhibited cell autophagy and EC dynamic alterations induced by galectin-3.CONCLUSION:These findings demonstrate that galectin-3 can induce an Akt signaling cascade leading to cell autoph-agy, and then the differentiation and angiogenesis of pulmonary artery endothelial cells .
作者： 刊期： 2016年第08期
目的：Mip2是心肌缺血后适应的一个分子靶点，其表达能抑制氧化应激诱导的心肌细胞凋亡。基于MIP2为WD蛋白，本科室冯衍生博士利用大鼠心肌缺血再灌注动物模型，对MIP2可能的相互作用蛋白进行了筛选，质谱鉴定了若干个蛋白质，其中包括VDAC，但VDAC包括VDAC1、VDAC2和VDAC3，它们与MIP2的关系尚不清楚。本研究在此基础上进一步深入探讨MIP2的心肌细胞保护机制。方法：首先构建了MIP2和VDAC真核表达载体，利用了基因共转染探讨MIP2与VDAC可能的相互作用；然后采用不同的抗体免疫共沉淀加Western blot免疫印迹技术，主要探讨MIP2与VDAC1的相互作用；利用免疫荧光定位探讨MIP2与VDAC1在H9c2细胞内的分布；采用MIP2的结构突变，研究MIP2与VDAC1相互作用的结构域；后通过MIP2的全长与突变体探讨其对H9c2心肌细胞膜电位与细胞死亡率的影响，观察MIP2及其与蛋白相互作用对氧化应激损伤心肌细胞的保护作用。结果：MIP2与VDAC基因共转染后免疫共沉淀加Western blot 鉴定，结果显示MIP2与VDAC1和VDAC2有相互作用关系，与VDAC3无相互作用；GFP与VDAC1抗体免疫共沉淀进一步证明了这种相互作用；细胞免疫共定位显示，MIP2与VDAC1分布于细胞同一区域，支持其在细胞中存在相互作用；MIP2结构突变显示，位于其C端的WD40是与其它蛋白相互作用的结构域。心肌细胞转染基因后施以氧化应激处理，结果显示，虽然MIP2全长能抑制氧化应激诱导的H9 c2心肌细胞线粒体膜电位降低和细胞死亡，但其蛋白结合结构域不能有效抑制这种诱导性的膜电位降低与细胞死亡。结论：VDAC1是MIP2的一个作用靶点，MIP2 C端的WD40是其与VDAC1相互作用的一个结构域。 MIP2能抑制氧化应激诱导的心肌细胞线粒体膜电位降低与细胞死亡，其机制可能与调节VDAC1有关。
作者：蒋磊；陈广斌；王浩；刘可；张华莉；肖献忠 刊期： 2016年第08期
目的：探讨抑制miR－21可否减轻CVB3诱导的BALB／c小鼠心脏微血管损伤，阻断致病因子向靶器官迁移，从而减轻靶器官组织病变。方法：3～4周龄雄性BALB／c小鼠CVB3腹腔注射后饲养1周诱导急性病毒性心肌炎（ VMC）模型；BALB／c小鼠每月腹腔注射CVB31次共饲养3个月，诱导为慢性VMC模型。注射CVB3同时尾静脉注射抗miR－21质粒以敲低miR－21表达。结果：急性VMC小鼠外周血miR－21表达增加，心肌Bcl－2和CVB3－VP1表达增加。小鼠体内注射anti－miR－21质粒后，外周血miR－21表达降低，心肌caspase－3活性和CVB3－VP1表达下降，Bcl－2表达增加，HE染色心肌组织及心脏微血管病变减轻，TUNEL染色心肌细胞凋亡减少。慢性VMC小鼠心肌胶原表达增加，微血管密度减少，心功能下降。敲低miR－21增加慢性VMC小鼠心肌微血管密度，减少胶原沉积，改善小鼠心功能。体外过表达miR－21诱导CMVECs凋亡，减少心脏微血管新生。结论：靶向抑制miR－21可能通过抑制CMVECs凋亡，阻断致病因子向靶器官迁移，降低心肌病毒载量、减轻心肌炎心肌病变。慢性VMC小鼠中，可能是通过减少胶原沉积、减轻心肌纤维化，增加微血管新生，改善心功能。因此，miR－21可能是治疗病毒性心肌炎的新靶点。
作者：虞勇；李冰玉；贾剑国；邹云增；陈瑞珍 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：探讨儿茶酚抑素（ CST）对间歇低氧高血压大鼠的作用及机制。方法：健康雄性SD大鼠随机分为3组：control组、IH （间歇低氧组）组和IH＋CST组（于低氧前3 d皮下埋植含CST 20 nmol? kg －1? d－1的微量渗透泵）。后2组置于间歇低氧舱中，舱内氧浓度为（5±0．5）％～（21±0．5）％，低氧－复氧循环时间为120 s（60 s＋60 s），8 h／d，共3周。颈总动脉插管测收缩压（SP）、舒张压（DP）和平均压（MP），检测血浆中氧化／抗氧化损伤指标，Western blot 法检测主动脉和肾组织中核因子E2相关因子2（Nrf2）蛋白表达的变化。结果：SP、DP及MP，IH组均比control 组高（P＜0．01），而IH＋CST 组则显著低于IH 组（P＜0．01）。 IH组的MPO和MDA含量显著高于control组（P＜0．05），而SOD和羟自由基抑制率显著低于control组（P＜0．01）；IH＋CST组的MPO和MDA明显低于IH组（P＜0．05），SOD和羟自由基抑制率显著高于IH组（P＜0．01）。与control组相比， IH组大鼠主动脉和肾组织胞浆、胞核中Nrf2蛋白的表达均显著下调（P＜0．05）；IH＋CST组与IH组相比，胞浆中Nrf2蛋白的表达显著下调（P＜0．05），而胞核中Nrf2蛋白的表达显著上调（P＜0．05）。结论：CST有减轻间歇低氧致大鼠高血压的作用，该作用可能与其通过Nrf2－ARE信号通路调节氧化应激反应有关。
作者：陈然；范小芳；郑青青；丁露；薛峰；王永煜；龚永生 刊期： 2016年第08期
目的：平滑肌蛋白22α（ smooth muscle protein 22α，SM22α）被视为是细胞衰老的标志物，但是其在血管平滑肌细胞（ vascu－lar smooth muscle cell ，VSMC）衰老过程中的作用尚不清楚。本研究旨在探讨SM22α在VSMC衰老和血管老化进程中的作用。方法：利用angiotensin II（Ang II，10－7 mol／L）慢性刺激诱导VSMC衰老；用野生型和SM22α基因敲除小鼠皮下植泵，持续灌注Ang II（1μg? kg－1? min－1）4周，复制高血压模型。通过敲低和过表达SM22α观察其对VSMC衰老及调控通路蛋白表达和活性的影响。结果：Ang II持续刺激可诱导VSMC衰老，伴随着SM22α的表达增高。敲低SM22α可减弱Ang II诱导的VSMC衰老，过表达则反之。在Ang II诱导VSMC衰老条件下，SM22α表达上调抑制Mdm2与p53的结合，上调p53含量。 SM22α表达增加抑制Akt与Mdm2的磷酸化活化，导致Mdm2与p53的结合减弱。 SM22α基因敲除改善Ang II诱导的主动脉VSMC衰老和血压升高。结论：SM22α表达上调抑制Akt／Mdm2通路激活，进而减弱Mdm2与p53的结合，上调p53的表达量，促进衰老。
作者：苗穗兵；谢肖立；尹亚娟；赵丽丽；舒亚南；陈荣；陈鹏；董丽华；林燕玲；吕品；张丹丹；聂茜；薛震颖；韩梅 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
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
