AREL1 resists the apoptosis induced by TGF-β by inhibiting SMAC in vascular endothelial cells

Abnormal apoptosis of vascular endothelial cells is an important feature of arteriosclerosis (AS). Here, we induced apoptosis in human umbilical vein endothelial cells (HUVECs) using transforming growth factor-β (TGF-β), and investigated the role of antiapoptotic E3 ubiquitin ligase (AREL1) in the apoptosis of vascular endothelial cells. We proved that AREL1 is downregulated in TGF-β treated HUVECs. The overexpression of AREL1 inhibits the activation of Caspase-3 and Caspase-9 and attenuates cell apoptosis induced by TGF-β. According to the result of coimmunoprecipitation, AREL1 interacts with the proapoptotic proteins the second mitochondria-derived activator of caspases (SMAC) in TGF-β treated HUVECs. In addition, miR-320b inhibits the expression of AREL1, and the overexpression of AREL1 attenuates the apoptosis induced by miR-320b mimics in HUVECs. In conclusion, AREL1 is downregulated by miR-320b. AREL1 overexpression inhibits TGF-β induced apoptosis through downregulating SMAC in vascular endothelial cells. Our study explores pathogenesis regulation mechanism and new biological therapeutic targets for vascular disease.     

Flow patterns and endothelial cell morphology in a simplified model of an artificial ventricle

The aim of this study was to delineate the flow patterns in a non-unidirectional flow field inside a ventricle-shaped cell culture chamber, and examine the resulting morphology and integrity of the endothelium in select regions of the monolayer. The chamber was perfused by pulsatile flow, and the coherent motion of the fluid was studied using flow visualization aided by image analysis. Four distinct flow patterns were discerned and examined: central jet, flow impingement, flow separation, and recirculating eddies. The influence of these patterns on endothelial cell morphology was assessed after 20 h of exposure to flow. There were no signs of damage to the endothelium in the jet region nor was there evidence of cell alignment with the flow. Yet, there were changes in cell morphology and cytoskeletal architecture as compared to control. By contrast, within the eddies where the flow was highly disturbed, there was apparent damage to the endothelium. Thus, exposure of cells to random velocity fluctuations in regions of quasi-static flow compromises the integrity of the monolayer. Identification of such sites and acquisition of the knowledge necessary to protect the cells from denudation will be valuable for the endothelialization efforts of cardiac prostheses.     

Enantioselective lipase-catalyzed ester hydrolysis: Effects on rates and enantioselectivity from a variation of the ester structure

In order to make a preliminary study of substituent effects on the rate and enantioselectivity obtained in esterolytic reactions catalyzed by a lipase from Candida rugosa, a series of racemic esters, derived from some α-alkyl and α-halo phenylacetic acids, were prepared. The reactions were studied at pH 6.0 and 50°C under which conditions uncatalyzed hydrolysis was relatively slow. Reaction samples were studied at different points of time by means of analytical chiral reversed-phase liquid chromatography, which permitted the simultaneous determination of product enantiomeric excess and of the degree of total ester hydrolysis. These data were then used to calculate initial rates as well as enantioselectivity. An increase of the steric bulk of the α-substituent was found to highly decrease the rate of the reaction. On the other hand, rates were higher for the p-nitrophenyl esters than for the corresponding 2-chloroethyl esters. Consistently, the enantioselectivity was found to be higher for the latter type of ester. The esters of the α-halo (bromo and chloro) phenylacetic acids gave mandelic acid as the final product. This was caused by a rapid solvolysis of the α-halo phenylacetic acid initially formed. © 1993 Wiley-Liss, Inc.     

Biocompatible scaffolds based on collagen and oxidized dextran for endothelial cell survival and function in tissue engineering

Angiogenesis is a vital step in tissue regeneration. Hence, the current study aimed to prepare oxidized dextran (Odex)/collagen (Col)-hydrogels with laminin (LMN), as an angiogenic extracellular matrix (ECM) component, for promoting human umbilical vein endothelial cell (HUVEC) proliferation and function. Odex/Col scaffolds were constructed at various concentrations and temperatures. Using oscillatory rheometry, scanning electron microscopy (SEM), and cell viability testing, the scaffolds were characterized, and then HUVEC proliferation and function was compared with or without LMN. The gelation time could be modified by altering the Odex/Col mass ratio as well as the temperature. SEM showed that Odex/Col hydrogels had a more regular three-dimensional (3D) porous structure than the Col hydrogels. Moreover, HUVECs grew faster in the Col scaffold (12 mg/mL), whereas the Odex (30 mg/mL)/Col (6 mg/mL) scaffold exhibited the lowest apoptosis index. Furthermore, the expression level of vascular endothelial growth factor (VEGF) mRNA in the group without LMN was higher than that with LMN, and the Odex (30 mg/mL)/Col (6 mg/mL) scaffold without LMN had the highest VEGF protein secretion, allowing the cells to survive and function effectively. Odex/Col scaffolds, with or without LMN, are proposed as a tissue engineering construct to improve HUVEC survival and function for angiogenesis.     

阿司匹林通过调节Hippo途径抑制大鼠颅内动脉瘤的形成和机制

摘要 目的：探讨阿司匹林通过调节Hippo途径抑制大鼠颅内动脉瘤形成的机制。方法：选择40只健康SD大鼠分为对照组（不做任何处理）、假手术组（暴露双侧肾动脉后支和左颈总动脉,但不结扎）、动脉瘤组（结扎双侧肾动脉后支和左颈总动脉后生理盐水灌胃）和阿司匹林组（动脉结扎后阿司匹林灌胃）,各10只。灌胃12周后检测血清炎性因子[肿瘤坏死因子-α（TNF-α）、单核细胞趋化蛋白-1（MCP-1）、白细胞介素-6（IL-6）、IL-10]、血管内皮损伤标志物[一氧化氮（NO）、内皮素-1（ET-1）、血管内皮生长因子（VEGF）]。处死大鼠后,检测动脉瘤大小、壁厚比、内腔面积和中膜变薄长度,检测动脉瘤血管组织中Yes相关蛋白（YAP）表达情况。结果：（1）动脉瘤组和阿司匹林组大鼠Willis环上有明显凸起,且阿司匹林组大鼠凸起明显小于动脉瘤组。阿司匹林组大鼠动脉瘤大小、内腔面积和中膜变薄长度均显著小于动脉瘤组,壁厚比显著大于动脉瘤组（P＜0.05）。（2）动脉瘤组和阿司匹林组大鼠血清TNF-α、MCP-1、IL-6水平显著高于对照组和假手术组,IL-10水平显著低于对照组和假手术组（P<0.05）;阿司匹林组大鼠血清TNF-α、MCP-1、IL-6水平显著低于动脉瘤组,IL-10水平显著高于动脉瘤组（P<0.05）。（3）动脉瘤组和阿司匹林组大鼠血清NO水平显著低于对照组和假手术组,ET-1和VEGF水平显著高于对照组和假手术组（P<0.05）;阿司匹林组大鼠血清NO水平显著高于动脉瘤组,ET-1和VEGF水平显著低于动脉瘤组（P<0.05）。（4）动脉瘤组和阿司匹林组大鼠YAP蛋白表达相对吸光值显著高于对照组和假手术组（P<0.05）;阿司匹林组大鼠YAP蛋白表达相对吸光值显著低于动脉瘤组（P<0.05）。结论：阿司匹林能够显著减轻颅内动脉瘤大鼠炎性反应,改善血管内皮功能,抑制颅内动脉瘤形成,这可能与阿司匹林调控Hippo信号通路有关。     

Effect of anoxia and reoxygenation on antioxidant enzyme activities in immortalized brain endothelial cells

Summary The effects of anoxia and reoxygenation on major antioxidant enzyme activities were investigatedin vitro in immortalized rat brain endothelial cells (RBE₄ cells). A sublethal anoxic period of 12 h was assessed for RBE₄ cells using the neutral red uptake test. Anoxia markedly influenced the specific activity of catalase and superoxide dismutase, with no major effect on glutathione peroxidase or glutathione reductase. After 24 h postanoxia, the superoxide dismutase activity modulated by the presence or absence of oxygen returned to control value. Damage and recovery of RBE₄ immortalized rat brain endothelial cells in culture after exposure to free radicals and other oxygen-derived species provides a usefulin vitro model to study anoxia-reoxygenation trauma at the cellular level.     

Stimulation of endothelial cell migration in culture by ladsin,a laminin-5-like cell adhesion protein

Summary Ladsin is a laminin-like cell-adhesive scatter factor with potent cell motility-stimulating ability and was purified from serum-free conditioned medium of a malignant human gastric adenocarcinoma cell line STKM-1. To test its possible role in tumor angiogenesis, we investigated its effect on primary culture of endothelial cells (human umbilical vein endothelial cells) and endothelial cell line ECV304 in this study. Cell adhesion and motility effects of ladsin were observed in both types of endothelial cells. In cell-attachment assay, ladsin interacted with integrin α3β1 that was expressed on the endothelial cell surface. In Boyden chambers, ladsin stimulated both directed and random migration of ECV304 cells. Ladsin induced repair of artificial wounds generated in ECV304 cell monolayers by stimulating cell migration. Ladsin did not affect the growth rate of ECV304 cells at a low cell density but significantly increased the saturation cell density. These results suggest that ladsin may be involved in the adhesion and migration of endothelial cells under some physiological and pathological conditions.     

Effect of moisture on lipase catalysed esterification of geraniol of palmarosa oil in non-aqueous system

Summary Optimisation of reaction conditions for the esterification of geraniol of palmarosa oil with n-butyric acid using immobilized lipase from Mucor miehei in non-aqueous system was carried out. Palmarosa oil could be easily esterified upto 95% w/w at 40°C in 24 h. Effect of moisture content was studied using Na₂SO₄ and recycling of the immobilized enzyme.     

Effects of sorbitol addition on the action of free and immobilized hydrolytic enzymes in organic media

The effect of the addition of sorbitol on the activity and stability of enzymes was examined by monitoring transesterification reactions performed in organic media at various water activities (a(w) = 0.08 to 0.97). Lipases from Chromobacterium viscosum and Candida rugosa immobilized on celite, and chymotrypsin, free or immobilized on celite, were used. When the sorbitol-containing enzymes were employed, higher reaction rates and less hydrolysis were observed. Immobilization of chymotrypsin resulted in high activity and operational stability, while the nonimmobilized enzyme was stable only in the presence of sorbitol. The activity of all preparations diminished after washing them with pyridine to remove sorbitol. Furthermore, severe stability problems occurred in the preparations lacking sorbitol. Sorbitol treatment, even after removal of the sorbitol itself, improved the activity of nonimmobilized chymotrypsin relative to the washed control. On the other hand, washing to remove sorbitol had a negative effect on the activity of both coimmobilized lipase and coimmobilized chymotrypsin. Addition of a substrate analogue, N-acetyl-L-phenylalanine, to chymotrypsin yielded a preparation that exhibited higher activity than both the control and its sorbitol-containing counterpart. Differential scanning calorimetry measurements revealed that the chymotrypsin-sorbitol complex was stable against thermal denaturation, undergoing transition at a high temperature (89 degrees C). The transition temperatures of the substrate-containing chymotrypsin and of the control were identical (72 degrees C). (c) 1995 John Wiley & Sons, Inc.