六味地黄方含药血清对氧化低密度脂蛋白损伤血管内皮细胞的保护作用

目的：观察六味地黄方大鼠含药血清对氧化低密度脂蛋白（ox-LDL）损伤血管内皮细胞（VEC）的保护作用。方法：以MTT法观察人脐静脉内皮细胞（HUVEC）的活力和流式细胞仪测定内皮细胞凋亡率,并通过测定胞内丙二醛（MDA）含量及细胞培养液中乳酸脱氢酶（LDH）、一氧化氮（NO）含量与超氧化物歧化酶（SOD）活力,探讨六味地黄方对血管内皮细胞损伤的保护作用及机制。结果：六味地黄方含药血清能促进ox LDL损伤的血管内皮细胞增殖、抑制ox LDL造成的内皮细胞凋亡、降低胞内MDA含量、减少细胞LDH释放量,提高SOD活力及NO含量。结论：六味地黄方对氧化低密度脂蛋白损伤的血管内皮细胞有一定的保护作用。     

Effect of Leptospira interrogans outer membrane proteins LipL32 on HUVEC

Leptospira cause disease through a toxin-mediated process by inducing vascular injury, particularly a small-vessel vasculitis. Breakdown of vessel endothelial cell integrity may increase vessel permeability which is correlated with the changes of tight junction and/or apoptosis in vessel endothelial cells. The specific toxin responsible remains unidentified. In this study, we amplified outer membrane protein LipL32 from the genome of Leptospira interrogans serovar Lai, and it was subcloned in pET32a(+) vector to express thioredoxin(Trx)–LipL32 fusion protein in Escherichia coli BL21(DE3). The protein was expressed and purified, and Trx–LipL32 was administered to culture with human umbilical vein endothelial cells (HUVEC) to elucidate the role of leptospiral outer membrane proteins in vessel endothelial cell. The purified recombinant protein was capable to increase the permeability of HUVECs. And the protein was able to decrease the expression of ZO-1 and induce F-actin in HUVECs display thickening and clustering. Moreover, apoptosis of HUVEC was significantly accelerated. But the fusion partner had no effect in these regards. It is possible that LipL32 is involved in the vessel lesions.     

Cell-derived microparticles contain caspase 3 in vitro and in vivo.

BACKGROUND: Microparticles (MP) from endothelial cells (endothelial microparticles; EMP) circulate in disease states, but the processes such as apoptosis or cell activation underlying their release are unclear. OBJECTIVES: We investigated whether adherent (viable) or detached (apoptotic) endothelial cells are the possible source of EMP in vitro, i.e. under control and interleukin (IL)-1alpha activation conditions, and in vivo. METHODS: Adherent and detached endothelial cells, and EMP, were isolated from human umbilical vein endothelial cell cultures (n = 6), treated without or with IL-1alpha (5 ng mL(-1); 24 h). Cell fractions were analyzed by flow cytometry for annexin V binding, propidium iodide (PI) and caspase 3 staining (n = 3). Caspase 3 in EMP was studied using Western blot (n = 6) and flow cytometry (n = 6). Plasma from healthy subjects and systemic lupus erythematosus patients (both n = 3) were analyzed for caspase 3-containing (E)MP. RESULTS: Detached but not adherent cells double-stained for annexin V and PI, confirming the apoptotic conditions of the detached cells and the viable nature of the adherent cells. Caspase 3 was solely present in the detached cells and procaspase 3 in the adherent cells. Caspase 3 was present in EMP from both control and IL-1alpha-treated cultures. Counts of EMP and detached cells, but not adherent cells, highly correlated (r = 0.959, P < 0.0001). In vivo circulating MP from nucleated (endothelial cells, monocytes) and anucleated cells (platelets, erythrocytes) contained caspase 3. CONCLUSIONS: EMP contain caspase 3 and may be mainly derived from detached (apoptotic) endothelial cells in vitro. The presence of caspase 3 in MP from anucleated cell types, however, suggests that its presence may not necessarily be related to apoptosis in vivo but may be associated with caspase 3 activation unrelated to apoptosis.     

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.     

腺苷对人静脉内皮细胞增殖的影响

目的探讨腺苷对人脐静脉内皮细胞（human umbilical vein endothelial cell，HUVEC）增殖的影响。方法MTT法观察腺苷的最佳作用浓度及最佳作用时问。结果4％FBS浓度下，腺苷具有明显的促HUVEC增殖作用，为最佳浓度。在10-^4 mol／L腺苷终浓度对细胞增殖有明显的促进作用，为最佳腺苷浓度。腺苷的促生艮作用在24h即出现，在48h已达到较高水平。结论在血供基本被阻断或血流基本正常的情况下，腺苷可能对细胞增殖尤影响；但在m流减少的情况下，腺苷的应用可以促进内皮细胞增殖，可能对血管新生有启动作用。     

Influence of carboxylic acid functionalization on the cytotoxic effects induced by single wall carbon nanotubes on human endothelial cells (HUVEC)

A vast variety of nanomaterials have been developed in the recent years, being carbon nanotubes (CNTs) the ones that have attracted more attention, due to its unique properties which make them suitable for numerous applications. Consequently, it is predicted that tons of CNTs will be produced worldwide every year, being its exposure of toxicological concern. Nanomaterials, once into the body, can translocate from the uptake sites to the blood circulation or the lymphatic system, resulting in distribution throughout the body. Thus, the vascular endothelium can be in contact with them and can suffer from their toxic effects. In this regard, the aim of this work was to investigate the cytotoxicity of single-walled carbon nanotubes (SWCNTs) on human endothelial cells evaluating the influence of acid carboxylic functionalization and also the exposure time (24 and 48 h). Biomarkers assessed were neutral red uptake, protein content, a tetrazolium salt metabolization and cell viability by means of the Trypan blue exclusion test. Cells were exposed to concentrations between 0 and 800 μg/mL SWCNTs for 24 and 48 h. Results have shown that both SWCNTs and carboxylic acid functionalized single-walled carbon nanotubes (COOH-SWCNTs) induce toxic effects in HUVEC cells in a concentration- and time-dependent way. Moreover, the carboxylic acid functionalization results in a higher toxicity compared to the SWCNTs.     

疡愈涂剂对人白细胞与血管内皮细胞黏附影响的分子机制

目的研究疡愈涂剂(YangYuTuJi,YYTJ)对白细胞与血管内皮细胞黏附影响的分子机制,探讨其促进皮肤创伤愈合的机制。方法用虎红染色法观察YYTJ高(132.5mg/L)、中(66.3mg/L)、低(33.2mg/L)浓度对中性粒细胞(polymorphonuclear,PMN)、人单核细胞(THP-1)与肿瘤坏死因子(tumor necrosis factor,TNF)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)黏附的影响,用荧光免疫细胞化学法观察YYTJ对HUVEC表面细胞间黏附分子-1(inter cell ularadhesion molecule-1,ICAM-1)、血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)和CD44表达的影响。结果 YYTJ和TNF共同作用HUVEC12h后,高浓度YYTJ能明显抑制HUVEC与PMN以及THP-1黏附(P<0.05),能明显抑制HUVEC表面ICAM-1和CD44表达(P<0.05)。中浓度YYTJ能明显抑制HUVEC表面VCAM-1和CD44表达(P<0.05,P<0.01)。低浓度YYTJ能明显降低HUVEC表面CD44表达(P<0.05)。结论疡愈涂剂通过降低TNF诱导的HUVEC表面黏附分子的表达,抑制白细胞与血管内皮细胞的黏附,从而抑制过度的炎性反应。