参芎葡萄糖注射液对H2O2诱导的HUVEC细胞氧化损伤的保护作用

目的:探讨参芎葡萄糖注射液(Shenxiong glucose injection,SGI)对H_2O_2诱导的人脐静脉上皮细胞(HUVEC)细胞氧化模型损伤的保护作用及其机制。方法:体外培养HUVEC人脐静脉内皮细胞,用H_2O_2(130 mmol·L~(-1))处理0.5 h,建立HUVEC细胞H_2O_2氧化损伤模型。SGI组HUVEC细胞用(6%,8%,10%)SGI预处理6 h后,再用H_2O_2处理0.5 h。用MTS法检测细胞存活率;用ELISA法检测乳酸脱氢酶(LDH)漏出量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活力;用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白质免疫印迹(Western blot)技术检测凋亡相关基因及蛋白B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax)和半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的mRNA和蛋白表达情况。结果:在130 mmol·L~(-1)H_2O_2作用细胞0.5 h的情况下,细胞存活率降低至50%左右,降低的程度合适,且实验结果重复性好,因此后续实验用该条件建立氧化损伤模型。与H_2O_2组比较,SGI预处理6 h能显著升高细胞存活率(P0.05,P0.01),减少LDH的外漏和MDA的生成(P0.05,P0.01),显著增加SOD,GSH-Px和CAT的活性(P0.05,P0.01)。RT-PCR和Western blot结果表明,SGI能显著上调Bcl-2的表达(P0.05,P0.01),下调Caspase-3,Bax的表达(P0.05,P0.01)。结论:参芎葡萄糖注射液能保护HUVEC细胞对抗H_2O_2诱导的氧化损伤,具有一定的剂量依赖关系,其作用机制可能与抑制细胞凋亡有关。     

牙龈卟啉单胞菌对脐静脉血管内皮细胞表达IL-8和MCP-1 mRNA的影响

目的：在转录水平上观察牙龈卟啉单胞菌对脐静脉血管内皮细胞（human umbilical vein endothelial cells,HUVECs）表达白细胞介素-8（IL-8）和单核细胞趋化蛋白-1（MCP-1）的影响。方法：厌氧培养Pg，原代培养HUVECs，RNA抽提，逆转录-聚合酶链式反应（RT-PCR）和mRNA比色定量法检测IL-8和MCP-1基因表达。结果：HUVECs基础表达IL-8和MCP-1mRNA,Pg以剂量依赖的方式增强IL-8和MCP-1mRNA的表达，Pg感染后1hIL-8mRNA开始增高，3h达到高峰，并持续到5h。而MCP-1的mRNA从Pg感染后1h开始增高，3h达到高峰，5h出现下降趋势。结论：Pg在转录水平上增强UHUVECs表达IL-8和MCP-1，这种上调作用可能是牙周病早期基因水平调控炎症细胞募集的重要因素，可能是牙周疾病的炎症反应和免疫反应中主要调控机制之一。     

杜鹃素对H_2O_2诱导的内皮细胞凋亡的保护作用研究

目的研究杜鹃素对H2O2诱导的人脐静脉内皮细胞(HUVEC)凋亡的保护作用。方法采用M TT法检测细胞活力,相差显微镜和荧光显微镜观察细胞形态变化。结果采用5、10、20μg/m L浓度的杜鹃素处理细胞后,内皮细胞活性呈浓度依赖性增加,细胞存活率提高;与H2O2组相比,细胞皱缩明显减少。杜鹃素20μg/m L可抑制H2O2诱导的人脐静脉内皮细胞凋亡。结论杜鹃素具有抑制H2O2诱导的人脐静脉内皮细胞凋亡的作用。     

狭鳕鱼皮胶原蛋白肽对过氧化氢诱导的人脐静脉内皮细胞氧化损伤的作用研究

目的：通过建立H2O2 导致人脐静脉内皮细胞损伤的模型来研究狭鳕鱼皮胶原蛋白肽作用。方法：用人脐静脉内皮细胞作为体外研究对象,采用不同的浓度（40M、200M、400M）胶原蛋白和维生素E预处理12h,然后在加入50uM H2O2继续培养12h造成损伤模型。模型成功后用CCK-8检测吸光度,根据试剂盒操作检测细胞内及培养上清液谷胱甘肽过氧化物酶（GXH-PX）,一氧化氮（NO）、过氧化氢酶（CAT）,丙二醛（MDA）等指标的含量和活性变化。结果：经胶原蛋白预保护后细胞增值率升高,GXH-PX、CAT等活性增强,NO含量升高,丙二醛含量降低。     

Biological activity of bevacizumab,a humanized anti-VEGF antibody <Emphasis Type="Italic">in vitro</Emphasis>

Bevacizumab (Avastin, Genentech) is a humanized monoclonal antibody targeting vascular endothelial growth factor (VEGF), a critical angiogenic factor involved in both physiological and pathological conditions. It has been recently approved by the US FDA as a first-line therapy for widespread metastatic colorectal cancer. This report is a detailed biological characterization of bevacizumab in a variety of in vitro models. It is shown that bevacizumab potently neutralizes VEGF and blocks its signal transduction through both the VEGFR-1 and VEGFR-2 receptors, as demonstrated by the inhibition of VEGF-induced cell proliferation, survival, permeability, nitric oxide production, as well as migration and tissue factor production. Although bevacizumab retains the ability to bind to human Fc receptors and complement protein C1q, it does not demonstrate cell or complement-mediated cytotoxicity in either VEGF producing or targeting cells. Thus the mechanism of anti-tumor activity of bevacizumab is most likely due to its anti-angiogenesis effect through binding and neutralization of secreted VEGF.     

17β-雌二醇对氧化型低密度脂蛋白由钙介导细胞骨架损伤的抑制作用

目的　通过观察 1 7β 雌二醇 (1 7β E2 )与氧化低密度脂蛋白 (OX LDL)作用于血管内皮细胞后细胞内微丝肌动蛋白的变化以及同细胞内钙离子变化间的关系 ,以探讨雌激素的血管保护作用机制。方法　采用ECV 30 4脐静脉内皮细胞株体外培养 ,分为空白对照组 ,OX LDL(2 0 0 μg/ml)组 ,4× 1 0 - 7mol/LE2 作用组 ,E2 保护组 ,采用激光扫描共聚焦显微镜分别观察OX LDL作用后 1 2h时的胞内钙离子的变化及OX LDL作用 2 4h时的细胞骨架微丝的改变。结果　OX LDL作用 1 2h后细胞内钙离子浓度明显升高 ,而细胞骨架微丝也在 2 4h后发生明显的破坏。雌激素预处理后可明显抑制OX LDL作用导致后细胞内钙离子浓度的升高 ,并且明显地防止细胞骨架微丝损伤的发生。结论　 1 7β E2 可能通过抑制OX LDL导致的细胞内钙离子升高从而防止细胞骨架微丝损伤的发生 ,这可能是雌激素产生血管保护作用的机制之一。     

A large mobility of hydrophilic molecules at the outmost layer controls the protein adsorption and adhering behavior with the actin fiber orientation of human umbilical vein endothelial cells (HUVEC)

Adhesion behaviors of human umbilical vein endothelial cells (HUVECs) are interestingly affected by the mobility of hydrophilic chains on the material surfaces. Surfaces with different molecular mobilities were prepared using ABA-type block copolymers consisting polyrotaxane (PRX) or poly(ethylene glycol) (PEG) central block (A block), and amphiphilic anchoring B blocks of poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate) (PMB). Two different molecular mobilities of the PRX chains were designed by using normal α-cyclodextrin (α-CD) or α-CD whose hydroxyl groups were converted to methoxy groups in a given ratio to improve its molecular mobility (PRX–PMB and OMe-PRX–PMB). The surface mobility of these materials was assessed as the mobility factor (Mf), which is measured by quartz crystal microbalance with dissipation monitoring system. HUVECs adhered on OMe-PRX–PMB surface much more than PRX–PMB and PMB-block–PEG–block-PMB (PEG–PMB) surfaces. These different HUVEC adhesions were correlated with the density of cell-binding site of adsorbed fibronectin. In addition, the alignment of the actin cytoskeleton of adhered HUVECs was strongly suppressed on the PEG–PMB, PRX–PMB, and OMe-PRX–PMB in response to the increased Mf value. Remarkably, the HUVECs adhered on the OMe-PRX–PMB surface with much less actin organization. We concluded that not only the cell adhesion but also the cellular function are regulated by the molecular mobility of the outmost material surfaces.     

Cellular senescence determines endothelial cell damage induced by uremia

Renal dysfunction is closely associated with endothelial damage leading to cardiovascular disease. However, the extent to which endothelial damage induced by uremia is modulated by aging is poorly known. Aging can render endothelial cells more susceptible to apoptosis through an oxidative stress-dependent pathway. We examined whether senescence-associated to oxidative stress determines the injury induced by the uremia in endothelial cells.     

Protein kinase C isoforms in atherosclerosis: Pro- or anti-inflammatory?

Atherosclerosis is a pathologic condition caused by chronic inflammation in response to lipid deposition in the arterial wall. There are many known contributing factors such as long-term abnormal glucose levels, smoking, hypertension, and hyperlipidemia. Under the influence of such factors, immune and non-immune effectors cells are activated and participate during the progression of atherosclerosis. Protein kinase C (PKC) family isoforms are key players in the signal transduction pathways of cellular activation and have been associated with several aspects of the atherosclerotic vascular disease. This review article summarizes the current knowledge of PKC isoforms functions during atherogenesis, and addresses differential roles and disputable observations of PKC isoforms. Among PKC isoforms, both PKCβ and PKCδ are the most attractive and potential therapeutic targets. This commentary discusses in detail the outcomes and current status of clinical trials on PKCβ and PKCδ inhibitors in atherosclerosis-associated disorders like diabetes and myocardial infarction. The risk and benefit of these inhibitors for clinical purposes will be also discussed. This review summarizes what is already being done and what else needs to be done in further targeting PKC isoforms, especially PKCβ and PKCδ, for therapy of atherosclerosis and atherosclerosis-associated vasculopathies in the future.     

Discovery of a New Series of Naphthamides as Potent VEGFR-2 Kinase Inhibitors

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