苦荞蛋白源肽AFYRW对脂多糖诱导的人脐静脉内皮细胞损伤的影响

目的：探讨苦荞蛋白源肽AFYRW在脂多糖（lipopolysaccharide,LPS）诱导的血管内皮细胞损伤中的作用。方法：采用LPS诱导人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs）建立血管内皮细胞炎症损伤模型;采用CCK8试剂盒检测AFYRW对细胞增殖活力的影响;Western blot检测血管细胞黏附分子-1(vascular cell adhension molecule-1,VCAM-1)、细胞内黏附分子-1(intercellular adhension molecule-1,ICAM-1)、白细胞介素（interleukin,IL）-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、内皮型一氧化氮合酶（endothelial nitric oxide synthase,e NOS）、诱导型一氧化氮合酶（inducible nitricoxide synthase,iNOS）水平及核因子-κB(nuclear factor kappa-B,NF-κB)p65的磷酸化水平;酶法...     

Impact of thermal variations on biochemical and physiological traits in Pectinatus sp

The influence of temperature on cellular fatty acid composition and on heat stress tolerance was studied in the two species of Pectinatus, an anaerobic gram-negative bacterium. Cellular fatty acid (FA) patterns were determined for Pectinatus species cultivated in MRS medium at various defined conditions of temperature and pH. Our study shows that fluctuations of growth temperature and pH induced important changes in the ratio of unsaturated FAs (UFAs) to saturated FAs (SFAs). The major differences in the FA composition as a function of growth temperature concerned C15:0 and C17:0 for the SFAs and C15:1 and C17:1 for the UFAs. The most significant adaptation of lipid composition to lower growth temperatures was the strong increase of UFAs, particularly for C15:1 and C17:1 concomitantly with a decrease of SFAs (C15:0 and C17:0). When the pH of the culture medium was lowered from 6.2 to 4.0, a notable drop in the synthesis of the UFAs C15:1 and C17:1 was observed together with an important increase of C18-cyclopropane (C18-cyc) and high carbon number SFAs. Thermal modifications also provoked changes in Pectinatus behaviour. We observed that P. cerevisiiphilus was more heat sensitive than P. frisingensis. Mild exponential phase cells were treated for 1 h, at 40 degrees C for P. cerevisiiphilus or at 41 degrees C for P. frisingensis. This thermal adaptation induced tolerance against heat challenge (49 and 50 degrees C for P. cerevisiiphilus and P. frisingensis, respectively). Survival of P. cerevisiiphilus and P. frisingensis adapted cells was, respectively, 3400- and 790-fold higher than control. Interestingly, adapted cells of P. cerevisiiphilus were more thermotolerant than P. frisingensis pretreated cells.     

Receptor-mediated uptake of hypertriglyceridaemic VLDL remnants by mouse macrophages in vitro

Following incubation of the VLDL isolated from hypertriglyceridaemic patients with normal human postheparin plasma VLDL remnants were obtained. 125I-VLDL remnants were taken up and catabolized by mouse macrophages in vitro. This was a receptor-dependent process. Unlabelled VLDL remnants added to the incubation medium inhibited the degradation of 125I-VLDL remnants in the cells. This could not be produced by the presence of VLDL or acetyl-LDL in the medium. It shows that mouse macrophages bind VLDL remnants by means of an other receptor than that for acetyl-LDL. The cell cholesterol content increased after incubation of the cells with VLDL remnants or acetyl-LDL, but not with native VLDL or LDL.     

Effects of flavonoids on the expression of the pro-inflammatory response in human monocytes induced by ligation of the receptor for AGEs

Increasing evidence has shown advanced glycation end products (AGEs) receptor ligation (RAGE) to be an important part of complex interactions of the oxidative stress and pro-inflammatory responses. In this study, flavonoids were used to monitor the protective effects against the oxidative damage and inflammation mediated by AGEs in human monocytes. S100B (RAGE ligand) treatment in human THP-1 monocytic cells (THP-1) significantly increased gene expression of the pro-inflammatory cytokines TNF-alpha and IL-1beta; chemokines MCP-1 and IP-10; adhesion factors platelet endothelial cell adhesion molecule (PECAM-1) and beta2-integrin; and pro-inflammatory cyclooxygenase-2 (COX-2). S100B treatment with quercetin and catechin in THP-1 cells had inhibitory effects on the expression of pro-inflammatory genes and protein levels. Quercetin and catechin could regulate S100B-activated oxidant stress-sensitive pathways through blocking p47phox protein expression. Treatment with quercetin and catechin could eliminate reactive oxygen species (ROS) to reduce oxidative stress stimulated by S100B in THP-1 cells. Quercetin and catechin also showed different regulatory abilities on mitogen-activated protein kinase (MAPK) signaling pathways by inhibiting protein expression in S100B-stimulated inflammatory responses in THP-1 cells. This study suggests that quercetin and catechin may be of benefit for diabetic vascular complications due to its antioxidant abilities against AGE-mediated oxidative stress through oxidative stress-sensitive and oxidative stress-responsive signaling pathways, which lead to inflammation in human monocytes.     

Effects of vitamin D3 on the expression of growth-related oncogene-α in THP-1 cells and human primary monocytes

Asthma and many autoimmune diseases, such as systemic lupus erythematosus, have been reported to associate with vitamin D deficiency recently. Growth-related oncogene-α (GRO-α)/CXCL1, a neutrophil-related chemokine, have an important influence on the chronic inflammation of these diseases. It is unknown whether vitamin D has regulatory effects on GRO-α expression in human monocytes. To this end, the human monocytic leukemia cell line, THP-1, and human primary monocytes were pretreated with 1α, 25-(OH)(2)D(3), and was stimulated with lipopolysaccharide (LPS). Supernatants were collected to determine GRO-α level by ELISA. The intracellular signaling was investigated by nuclear factor (NF)-κB inhibitor, the mitogen-activated protein kinase (MAPK) inhibitors, and Western blot. In our studies, LPS-induced GRO-α was significantly enhanced in THP-1 cells, but suppressed in human primary monocytes by 1α, 25-(OH)(2)D(3). Western blotting revealed that 1α, 25-(OH)(2)D(3) increased LPS-stimulated pp38 expression in THP-1 cells, but suppressed LPS-stimulated pMEK1/2-pERK and pJNK in human primary monocytes. In conclusion, the opposite effects of 1α, 25-(OH)(2)D(3) on GRO-α expression in THP-1 cells and human primary monocytes indicated that the data from THP-1 cells should be further confirmed by human primary monocytes. Moreover, vitamin D3 may have potentiality in treating GRO-α-related chronic inflammatory diseases, like asthma and autoimmune diseases.     

榆干离褶伞溶栓酶对氧化应激损伤人脐静脉内皮细胞的保护作用

目的：探讨榆干离褶伞（Lyophyllum ulmarium,L.u）溶栓酶对H2O2诱导人脐静脉内皮细胞（humanumbilical vein endothelial cell,HUVEC）损伤的保护作用。方法：用H2O2诱导HUVEC氧化损伤,采用四甲基偶氮唑蓝（methyl thiazolyl tetrazolium,MTT）法观察细胞存活率;用流式细胞仪检测细胞线粒体跨膜电位（ΔΨm）和总活性氧（reactive oxygen species,ROS）水平;Western blotting法检测Caspase-8、Caspase-9、Caspase-3等蛋白的表达水平。结果：L.u溶栓酶可显著提高HUVEC的存活率,明显抑制H2O2诱导的细胞内ROS生成和线粒体跨膜电位的下降,并能降低Caspase-8、Caspase-9和Caspase-3的表达水平。结论：L.u溶栓酶对氧化应激损伤的HUVEC有保护作用,其机制可能与抑制细胞凋亡有关。     

The nutritional supplement Active Hexose Correlated Compound (AHCC) has direct immunomodulatory actions on intestinal epithelial cells and macrophages involving TLR/MyD88 and NF-κB/MAPK activation

Active Hexose Correlated Compound (AHCC) is an immunostimulatory nutritional supplement. AHCC effects and mechanism of action on intestinal epithelial cells or monocytes are poorly described. AHCC was added to the culture medium of intestinal epithelial cells (IEC18 and HT29 cells) and monocytes (THP-1 cells) and assessed the secretion of proinflammatory cytokines by ELISA. Inhibitors of NFκB and MAPKs were used to study signal transduction pathways while TLR4 and MyD88 were silenced in IEC18 cells using shRNA. It was found that AHCC induced GROα and MCP1 secretion in IEC18 and IL-8 in HT29 cells. These effects depended on NFκB activation, and partly on MAPKs activation and on the presence of MyD88 and TLR4. In THP-1 cells AHCC evoked IL-8, IL-1β and TNF-α secretion. The induction of IL-8 depended on JNK and NFκB activation. Therefore, AHCC exerts immunostimulatory effects on intestinal epithelial cells and monocytes involving TLR4/MyD88 and NFκB/MAPK signal transduction pathways.     

Arabinogalactan-type polysaccharides (APS) from Cordyceps militaris grown on germinated soybeans (GSC) induces innate immune activity of THP-1 monocytes through promoting their macrophage differentiation and macrophage activity

In this study, the immunemodulatory activities of arabinogalactans-type polysaccharide (APS) from Cordyceps militaris grown on germinated soybeans was evaluated in THP-1 human monocytes. The cellular size and the number of intracellular organelles of THP-1 monocytes were increased by APS. Also, APS-treated THP-1 cells significantly developed cellular adherence and macrophagic differentiation to the culture plate surface. APS noticeably enhanced phagocytic activity of THP-1 cells against IgG-FITC-latex beads. APS induced the level of TNF-α, IL-12 p40, and IL-8 as well as TLR2 and TLR4 mRNAs. APS can be developed as a promising immunmodulating agent with macrophage-activating properties.     

Oxidative stress, genotoxicity, and vascular cell adhesion molecule expression in cells exposed to particulate matter from combustion of conventional diesel and methyl ester biodiesel blends

Our aim was to compare hazards of particles from combustion of biodiesel blends and conventional diesel (D(100)) in old and improved engines. We determined DNA damage in A549 cells, mRNA levels of CCL2 and IL8 in THP-1 cells, and expression of ICAM-1 and VCAM-1 in human umbilical cord endothelial cells (HUVECs). Viability and production of reactive oxygen species (ROS) were investigated in all cell types. We collected particles from combustion of D(100) and 20% (w/w) blends of animal fat or rapeseed oil methyl esters in light-duty vehicle engines complying with Euro2 or Euro4 standards. Particles emitted from the Euro4 engine were smaller in size and more potent than particles emitted from the Euro2 engine with respect to ROS production and DNA damage, but similarly potent concerning cytokine mRNA expression. Particles emitted from combustion of biodiesel blends were larger in size, and less or equally potent than particles emitted from combustion of D(100) concerning ROS production, DNA damage and mRNA of CCL2 and IL8. ICAM-1 and VCAM-1 expression in HUVECs was only increased by D(100) particles from the Euro4 engine. This suggests that particle emissions from biodiesel in equal mass concentration are less toxic than conventional diesel.     

AngⅡ诱导人脐静脉内皮细胞建立高血压损伤模型

目的:采用高血压发病因子血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)建立高血压损伤模型。方法:使用不同浓度(0.01、0.10、1.00μmol/L和10.00μmol/L)AngⅡ诱导HUVECs不同时间(3、6、9、12 h和24 h),通过考察HUVECs形态、活性、功能、微观结构及凋亡程度来评价HUVECs损伤程度,同时采用交叉设计方差分析和多重比较方法得到AngⅡ最适诱导浓度和最佳诱导时间。结果:AngⅡ呈浓度依赖式诱导HUVECs损伤,最佳诱导条件为1.00μmol/L AngⅡ诱导12 h,此时HUVECs活性为44.85%、NO含量为43.57μmol/L、总一氧化氮合酶活力为6.99 U/mg pro、内皮型一氧化氮合酶活力为1.89 U/mg pro、丙二醛含量为7.46 nmol/mL、超氧化物歧化酶活力为27.29 U/mg pro、细胞凋亡率为41.5%,微观结构显示核膜皱缩,核仁消失,胞浆内出现大量空泡状结构,线粒体或细小或肿胀,粗面内质网扩张数目较少,部分核糖体丢失,平面内质网扩张,但细胞未解体,仍然保持细胞结构。结论:1.00μmol/L AngⅡ诱导12 h可以成功诱导HUVECs建立高血压损伤模型。     

Fatty liver in dairy cows post partum is associated with decreased concentration of plasma triacylglycerols and decreased activity of lipoprotein lipase in adipocytes

Cholesterol and phospholipid concentrations in serum lipoproteins, plasma activities of lecithin:cholesterol acyltransferase (LCAT) and phospholipid transfer protein (PLTP) and lipoprotein lipase (LPL) activity in adipose tissue biopsies were measured ante and post partum in dairy cows given either free or restricted access to feed during the dry period. After parturition, all cows were fed ad libitum. The purpose of this study was to try to understand the earlier observed marked drop post partum in plasma triacylglycerol (TAG) in terms of lipoprotein metabolism in cows developing fatty liver post partum. As would be expected, free access to feed during the dry period induced a rise of hepatic TAG concentrations post partum associated with a decrease in plasma TAG levels. Total and free cholesterol concentrations in the VLDL, IDL, LDL and HDL2 fractions fell immediately after parturition. VLDL and IDL cholesterol concentrations remained at a constant, low level during the entire sampling period post partum, whereas the drop in LDL and HDL2 cholesterol post partum was followed by a rebound rise. Plasma LCAT and PLTP activities decreased by on average 19% and 33%, respectively, after parturition and then rose to values seen before parturition, but there was no effect of feeding regimen during the dry period. Activities of LCAT and PLTP were significantly correlated with cholesterol and phospholipid concentrations in LDL and HDL2. Plasma LCAT activity, as measured with exogenous substrate, and PLTP activity were both positively correlated with HDL3 phospholipid levels. LPL activity in adipose tissue dropped after parturition, the drop being smaller after feeding ad libitum during the dry period. It is concluded that the drop in adipose tissue LPL activity post partum is at variance with the simultaneous fall in plasma TAG. Possibly, the decrease in adipose tissue LPL activity helps to channel fatty acids away from adipose tissue into the udder. The post-partum changes in lipid transfer proteins in the blood are in line with the changes observed in the levels of the lipoproteins.     

Antiangiogenic activity of indole-3-carbinol in endothelial cells stimulated with activated macrophages

The effect of indole-3-carbinol (I3C), a major indolic metabolite in cruciferous vegetables, on lipopolysaccharide (LPS)-activated macrophage-induced tube formation and its associated factors in endothelial EA hy926 cells was investigated. LPS significantly enhanced the capillary-like structure of endothelial cells (ECs) co-cultured with macrophages, but no such effect was observed in single-cultured ECs. I3C, on the other hand, suppressed such enhancement in concert with decreased secretions of vascular endothelial growth factor (VEGF), nitric oxide (NO), interleukin-6 (IL-6), and matrix metalloproteinases (MMPs). The results obtained from cultivating ECs with conditioned medium (CM) collected from macrophages suggested that both ECs and macrophages were inactivated by I3C. These results indicate that I3C from cruciferous vegetables may possess potential roles in preventing inflammation-associated angiogenic diseases.     

大黄酸对正常和氧化损伤内皮细胞的增殖和氧化保护作用

目的：观察大黄酸对正常和氧化损伤的人脐静脉血管内皮细胞(HUVEC)的影响。方法：建立H2O2氧化损伤模型,采用不同浓度的大黄酸对内皮细胞进行处理,检测内皮细胞乳酸脱氢酶(LDH)释放率、丙二醛(MDA)含量、一氧化氮(NO)含量、一氧化氮合酶(NOS)活力和超氧化物歧化酶(SOD) 活力及谷胱甘肽过氧化物酶(GSH-Px) 活力的变化;采用流式细胞术研究内皮细胞的凋亡情况,探讨大黄酸对内皮细胞的保护作用。结果：大黄酸浓度在16μmol/L以下时,对HUVEC的增殖无显著性影响;浓度在2μmol/L以上时,减轻HUVEC受到氧化损伤的程度,对氧化性损伤具有保护作用;同时降低了LDH释放率,下调了MDA含量,提高了NO含量和NOS、SOD、GSH-Px的酶活力,显著减少了细胞凋亡率。结论：HUVEC经2～16μmol/L大黄酸处理,可以有效减轻H2O2对其造成的氧化损伤,维持正常的生理形态。     

Cover Picture – Mol. Nutr. Food Res. 5/2008

Cancer chemoprevention and hemotherapy: dietary polyphenols and signalling pathways Inhibitory effects of trans‐resveratrol analogues on human colon tumoral cells Potential of sphingomyelin as a chemopreventive gent in colon cancer Unsaturated fatty acids liberated from VLDL cause apoptosis in endothelial cells     

Trans C18:1通过NOS-NO系统诱导内皮细胞损伤研究

为了观察trans C18:1对人脐静脉内皮细胞损伤的影响及其与NOS-NO系统的关系,首先将不同浓度trans C18:1 (50、100、200、400μmol/L)与人脐静脉内皮细胞共培养24或48h后,MTT法检测细胞存活率;用trans C18:1(200μmol/L)处理内皮细胞24h后,分别检测一氧化氮含量(NO)及一氧化氮合酶(NOS)活性;将trans C18:1与一氧化氮合酶阻断剂亚硝酸左旋精氨酸甲酯(LNAME)及一氧化氮供体(SNP)单一或联合处理内皮细胞,检测细胞存活率的变化。结果显示：trans C18:1以剂量和时间依赖方式导致内皮细胞的存活率下降; LNAME与trans C18:1联合处理内皮细胞后细胞存活率下降,而SNP与trans C18:1联合处理后细胞存活率上升;trans C18:1可诱导NO水平和内皮型一氧化氮合酶(eNOS)活性显著下降,而诱导型一氧化氮合酶(iNOS)活性无显著改变。表明：trans C18:1能通过抑制eNOS活性减少NO的分泌,并暗示NOS-NO系统可能是trans C18:1诱导内皮细胞损伤的作用机制之一。     

Localisation of C=C Bond and absolute quantification of unsaturated Fatty Acids in Vegetable Oils based on photochemical derivatisation reaction coupled with mass spectrometry

In this study, a strategy based on photochemical derivatisation and stable isotope labelling coupled with mass spectrometry (MS) for rapid and accurate identification and quantification of unsaturated fatty acids (UFAs) in vegetable oils was developed. Acetone was employed to label double bond(s) of UFAs via Paternò–Büchi (PB) reaction. The acetone-labelled UFAs could produce diagnostic ions specific to the locations of carbon–carbon double bond (C=C) in collision-induced dissociation (CID), which enabled confident assignment of C=C locations in UFAs. Furthermore, absolute quantification was achieved by using d6-acetone-labelled UFAs as internal standard to minimise the quantitation deviation due to nonuniformity response and ion suppression effects. By exploiting this method, precise identification and quantification of UFAs in nine kinds of vegetable oils including three kinds of Chinese special vegetable oils were achieved. This method exhibits a powerful potential use for the localisation of C=C bond(s) and quantitative analysis of UFAs, which is valuable for discovery of unusual UFAs in special oils and crops, and quality control of edible oils.     

榆干离褶伞发酵液对血管内皮细胞的保护作用

为了探讨榆干离褶伞(Lyophyllum ulmarium,简写为LU)发酵液对人脐静脉内皮细胞(HUVEC)的保护作用,分别用20、40、80 mg/L LU干预HUVEC后用过氧化氢诱导凋亡,采用MTT法观察LU发酵液对细胞存活率的影响;通过比色法测定细胞上清液乳酸脱氢酶(LDH)活性;用流式细胞仪检测细胞凋亡率;Western blot法检测bcl-2、bax、Caspase-3、Caspase-9蛋白质的表达。结果显示:LU发酵液显著提高内皮细胞的存活率,降低细胞上清液LDH活性,抑制H2O2诱导的细胞凋亡,降低bax、Caspase-3、Caspase-9的表达,提高bcl-2的表达。这说明LU发酵液对血管内皮细胞有保护作用,其机制可能与抑制凋亡有关。     

Protective effect of Cordyceps militaris against high glucose-induced oxidative stress in human umbilical vein endothelial cells

The protective effect of Cordyceps militaris against high glucose-induced oxidative stress in human umbilical vein endothelial cells (HUVECs), as compared with Cordyceps sinensis, was examined. The cytotoxicity of HUVECs induced by 40 mM glucose was ameliorated by water extracts of C. militaris (CME) and water extracts of C. sinensis (CSE). CME and CSE inhibited the increase in ROS and NO in HUVECs induced by 40 mM high glucose. Moreover, CME increased the Bcl-2/Bax ratio, modulated the mitochondrial membrane potential and reduced the caspase-3 activity in high glucose-induced HUVECs. In addition, cordycepin, a component of CME and CSE, displayed protective effects against oxidative stress, which was partly responsible for the cytoprotective effects of CME and CSE against high glucose-induced oxidative stress in HUVECs. Overall, the obtained results show C. militaris helps preventing diabetic endothelial dysfunction and related complications.