己酮可可碱与蛋白激酶C抑制剂对佛波醇酯诱导脑微血管内皮细胞表达细胞间粘附分子-1的影响

目的:研究佛波醇酯(PMA)诱导大鼠脑微血管内皮细胞(RBMEC)表达细胞间粘附分子-1(ICAM-1)及PKC抑制剂H7与己酮可可碱(PTX)的抑制作用。方法:采用ELISA方法测定培养RBMEC表达ICAM-1.结果:PMA在10-100nmol·L~(-1)范围内剂量依赖性地诱导RBMEC表达ICAM-1;在4-16h范围内时间依赖性诱导RBMEC表达ICAM-1。H7和PTX分别在5-50μmol·L~(-1)和1-100μmol·L~(-1)范围内剂量依赖性抑制PMA诱导的RBMEC表达ICAM-1。PTX 100μmol·L~(-1),H7 50μmol·L~(-1)时,抑制作用达最大[吸光度分别从(0.410±0.014)降至(0.175±0.022)和(0.182±0.013),P<0.01]。结论:PKC抑制剂及己酮可可碱能抑制PMA诱导RBMEC表达ICAM-1,表明PKC参与RBMEC ICAM-1表达调控。     

已酮可可碱对重症急性胰腺炎大鼠肺损伤的保护作用

目的 研究已酮可可碱(PTX)对重症急性胰腺炎(SAP)大鼠肺损伤的保护作用.方法 健康成年雄性SD大鼠72只,随机分为对照(C)组、实验(E)组和干预(P)组.E组和P组用5%牛磺胆酸钠逆行胰胆管注射制备SAP模型.然后,P组立即腹腔注射PTX.于制模后2、6和12 h观察胰腺和肺组织的病理学改变;ELISA法测定血清中性粒细胞趋化因子(CINC)和白细胞介素6(IL-6)水平变化;Western blot法检测肺组织核因子抑制蛋白(IκBα)的表达量.结果 E组血清CINC、IL-6水平及肺组织IκBα蛋白的表达均显著高于同时段C组(P<0.05);与E组比较,P组血清CINC、IL-6水平和肺组织IκBα蛋白表达量均明显降低(P<0.05),胰腺和肺组织病理改变减轻.结论 SAP肺损伤时,肺组织IκBα明显升高,核因子κB(NF-κB)活化,从而启动CINC、IL-6等过量表达.PTX可以抑制NF-κB/IκB信号通路的激活,对SAP肺损伤具有保护作用.     

蛋白激酶Cδ亚型抑制剂对脂多糖诱导肺微血管内皮细胞损伤的影响

目的研究蛋白激酶C(PKC)的亚型PKCδ在大鼠肺微血管内皮细胞(RPMVEC)中的表达及其抑制剂Rottlerin对脂多糖(LPS)导致RPMVEC单层通透性增高的影响。方法取体外培养的RPMVEC进行PKCδ的W estern b lot和免疫组化检测,用针头式滤器检测LPS及LPS+Rottlerin干预后RPMVEC单层滤过系数(K f)的变化。结果PKCδ在RPMVEC中有广泛的表达,其表达部位主要位于胞质。Rot-tlerin能减低LPS导致的RPMVEC单层通透性增高。结论LPS导致RPMVEC损伤的机制与PKC的激活有关,PKCδ亚型抑制剂可减轻LPS诱导的RPMVEC单层通透性增高。     

血管内皮细胞生长因子在脑星形细胞瘤的表达及其与微血管数的关系

目的：研究血管内皮生长因子（VEGF）在脑星形细胞瘤中的表达,探讨VEGF与肿瘤的病理分级、部位、体积、预后的关系,及其与微血管数的关系。方法：应用免疫组化法对60例脑星形细胞瘤的VEGF及微血管数进行检测,结合临床进行定性、定量、定位分析。结果：脑星形细胞瘤中VEGF高表达,并与微血管相关,VEGF的表达强度及微血管数与肿瘤的病理分级、生存率及某些部位有相关性,而与肿瘤的体积无相关性。结论：VEGF在脑星形细胞瘤生血管生成中起重要作用,VEGF的表达与微血管数均可做为判断肿瘤预后的重要参考指标。     

蛋白激酶C抑制剂和钙调素抑制剂对小鼠腹腔巨噬细胞释放肿瘤坏…

蛋白激酶Ｃ（ＰＫＣ）抑制剂Ｈ－７（０．３１－５０μｇ·ｍｌ＾－１）和钙调素（ＣａＭ）拮抗剂Ｘ－７（１０－４０００ｎｇ·ｍｌ＾－１）皆可浓度依赖地抑制脂多糖诱导卡西霉素启动的小鼠腹腔巨噬细胞释放肿瘤坏死因子（ＴＮＦ）。Ｗ－７（１０ｎｇ·ｍｌ＾－１）抑制ＴＮＦ释放的最佳时间为８ｈ。提示巨噬细胞释放ＴＮＦ可能依赖于ＰＫＣ和ＣａＭ。     

肿瘤坏死因子对牛肺动脉内皮细胞的损伤及蛋白激酶C抑制剂的抑制作用

研究表明:肿瘤坏死因子(TNF)可剂量依赖地引起牛肺动脉内皮细胞乳酸脱氢酶释放率(LDH%)升高,促进中性粒细胞向内皮细胞粘附,并可抑制内皮细胞增殖和DNA合成。蛋白激酶C(PKC)抑制剂1-(5-异喹啉磺酰基)-2-甲基哌嗪(H-7)和槲皮素一方面可剂量依赖地抑制TNF对内皮细胞的直接损伤,另方面又可通过抑制TNF诱导的中性粒细胞对内皮细胞粘附增加,减轻TNF对内皮细胞的间接损伤作用,同时还可抑制TNF对内皮细胞增殖和DNA合成的影响,从而间接加强内皮细胞对损伤的自我修复。     

丹酚酸B对缺氧损伤心脏微血管内皮细胞细胞间粘附分子表达的影响

目的 :探讨丹参有效成分丹酚酸B在心脏微血管内皮细胞 (CMEC)缺氧 /复氧损伤中的保护作用。方法 :通过心脏微血管内皮细胞体外培养技术 ,建立缺氧预适应 (HPC)模型并给予丹酚酸B ,应用酶联免疫方法检测细胞间粘附分子 1(ICAM 1)表达。结果 :在CMEC缺氧 3h ,复氧 6h后ICAM 1表达升高 ;缺氧预适应和应用丹酚酸B对CMEC进行预处理则能有效降低CMECICAM 1的表达。结论 :丹酚酸B在CMEC缺氧 /复氧损伤中可产生与缺氧预适应类似的保护效果。     

Motorcycle exhaust particles up-regulate expression of vascular adhesion molecule-1 and intercellular adhesion molecule-1 in human umbilical vein endothelial cells

Epidemiological studies have shown that there is a strong correlation between atherosclerosis and ambient air pollution. In this study, we found that motorcycle exhaust particles (MEP) induced adhesion between cells of the human monocytic leukemia cell line (THP-1) and human umbilical vein endothelial cells (HUVECs) in a time-and dose-dependent manner. In addition, MEP treatment induced both mRNA and protein expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in HUVECs. The IκB degradation and p65 nuclear translocation was found in MEP-treated HUVECs, suggested the involvement of nuclear factor-κB (NF-κB). MEP-induced VCAM-1 and ICAM-1 protein expression was inhibited by NF-κB inhibitor BAY 11-7085. Oxidative stress was also involved in the signaling of VCAM-1 and ICAM-1 expression. MEP treatment caused hydrogen peroxide and superoxide formation. Pretreatment with α-tocopherol could inhibit MEP-induced reactive oxygen intermediates generation and suppressed MEP-induced IκB degradation and adhesion molecules expression. Furthermore, the carbon black (CB) nanoparticles with different diameters could induce VCAM-1 and ICAM-1 protein expression; however, polycyclic aromatic hydrocarbons (PAHs) only increased the expression of ICAM-1 but not that of VCAM-1 in HUVECs. In this study, we found that MEPs could induce ICAM-1 and VCAM-1 expression through oxidative stress and NF-κB activation in HUVECs.     

Protein kinase C beta inhibitor LY333531 attenuates intercellular adhesion molecule-1 and monocyte chemotactic protein-1 expression in the kidney in diabetic rats

In vitro studies have shown that activation of protein kinase C (PKC) is a key mediator of intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) in a range of cell types and in response to high glucose, however, its role in the in vivo setting has not been clearly delineated. Streptozotocin-induced diabetic rats were treated with the PKC-beta isoform inhibitor LY333531 for 8 weeks. LY333531 treatment significantly attenuated increased urinary albumin excretion rate and glomerular volume and tubulointerstitial injury index as well as elevated PKC activity and PKC-beta protein expression in the kidney. Level of malondialdehyde was markedly higher and antioxidant enzyme activity such as superoxide diamutase and catalase as well as glutathione peroxidase were significantly lower in the kidney from diabetic rats than that of the control group. LY333531 administration could remit these changes. Increased macrophages recruitment as well as ICAM-1 and MCP-1 protein expression in the kidney were significantly inhibited by LY333531 in diabetic rats. It is concluded that mechanism of renoprotection of LY333531 may be correlated, at least partly, with suppression of increased macrophages recruitment and overexpression of ICAM-1 and MCP-1 in diabetic rats.     

C -reactive protein induced expression of adhesion molecules in cultured cerebral microvascular endothelial cells

AIM lsehemic stroke can trigger an acute phase response resulting in a rise of plasma concentration of C - reactive protein （CRP）. Clinical data about the relationship between CRP and prognosis suggest that CRP might be involved in the pathogenesis of cerebral ischemia. To determine the possible role of CRP in ischemic stroke, we performed a dose - dependent experiment in mouse brain microvascular endothelial cells （bEnd. 3 cells） with emphasis on its relation to cell adhesions molecules.     

低分子量肝素对白细胞黏附和黏附分子表达的影响

目的研究低分子量肝素对白细胞与血管内皮细胞体内外黏附和细胞间黏附分子 1(ICAM 1)表达的影响。方法倒置显微镜观察在致敏豚鼠肠系膜微循环中白细胞的滚动和黏附情况 ,体外观察大鼠中性粒细胞对培养的脐静脉内皮细胞的黏附情况 ,流式细胞仪测定ICAM 1的表达。结果低分子量肝素 2 0、4 0u kg对白细胞黏附有明显抑制作用 ;0 .5、1.0u ml(终浓度 )对中性粒细胞体外黏附有明显抑制作用 ;0 .12 5、0 .2 5、0 .5、1.0u ml对脐静脉内皮ICAM 1的表达有明显抑制作用。结论低分子量肝素有明显抑制白细胞黏附及降低ICAM 1表达的作用。     

The role of haem oxygenase-1 in the decrease of endothelial intercellular adhesion molecule-1 expression by curcumin

Intercellular adhesion molecule-1 (ICAM-1) is involved in neutrophil transmigration across endothelium during sepsis-induced acute lung injury and anti-ICAM-1 interventions may represent new strategy of pulmonary protection. Haem oxygenase-1 (HO-1) has been demonstrated to exert anti-inflammatory actions via decrease of expression of adhesion molecules. We investigated the role of HO-1 in the action of curcumin, a naturally occurring yellow pigment isolated from plant Curcuma longa L., on ICAM-1 expression in tumour necrosis factor-alpha-stimulated EA.hy926 cells and lungs of lipopolysaccharide-treated mice. Both, in vitro and in vivo curcumin induced HO-1 and curcumin-elicited induction of HO-1 was associated with inhibition ICAM-1 expression. Moreover, curcumin significantly inhibited pulmonary sequestration of leucocytes in response to lipopolysaccharide as evidenced by decrease of myeloperoxidase activity in lung tissue. Both in vitro and in vivo effects of curcumin were reversed by an inhibitor of HO activity, chromium (III) mesoporphyrin IX chloride. We conclude that induction of HO-1, via decrease of endothelial ICAM-1, plays a pivotal role in curcumin-dependent prevention of pulmonary sequestration of neutrophils in a mouse model of endotoxaemia.