肿瘤坏死因子α对肾小管上皮细胞ICAM—1表达的调控

目的：研究肿瘤坏死α（TNFα）对大鼠肾小管上皮细胞表达细胞间粘附分子－1（ICAM－1）的调控作用。方法：用不同浓度（1、10、50ng/ml)TNFα分别与大鼠肾小管上皮细胞（NRK－52E细胞）共同孵育1、4、12、24小时，用蛋白质印迹法（Western Blot)和逆转录聚合酶链反应技术（RT－PCR）分别观察ICAM－1的蛋白质和mRNA的表达水平。结果：正常对照组的肾小管上皮细胞低水平表达ICAM－1，在实验组中TNFα呈剂量依赖地诱导肾小管上皮细胞的ICAM－1的蛋白质和基因表达上调，蛋白质表达高峰在4小时，mRNA表达高峰在12小时。结论：肿瘤坏死因子α（TNFα）可能通过诱导肾小管上皮细胞表达ICAM－1增强而参与肾小管间质的免疫炎症反应。     

二甲双胍对饱和脂肪酸诱导的大鼠H9C2型心肌细胞损伤的保护作用研究

目的 探讨二甲双胍(Met)对饱和脂肪酸所诱导的大鼠H9C2型心肌细胞损伤作用的保护机制.方法 在对照、棕榈酸(PA)及3种不同浓度梯度Met与PA联合组培养液中培养大鼠H9C2型心肌细胞株24 h.采用蛋白质印迹法(Western blot)测定各组细胞核因子κB(NF-κB)p65、细胞间黏附因子(ICAM1)、磷酸化核因子κB抑制蛋白(p-IκBα)及磷酸化腺苷酸活化蛋白激酶(p-AMPK)的蛋白表达,实时荧光定量PCR测定各组细胞NF-κB、单核细胞趋化因子(CCL2)和ICAM1的mRNA表达.结果 与对照组相比,PA组的细胞中NF-κB p65、ICAM1、p-IκBα蛋白表达量增加(P<0.05);与PA组相比,Met+PA联合组细胞NF-κB p65、ICAM1、p-IκBα的蛋白表达量随Met浓度梯度增加表现为不同程度递减(P<0.05),p-AMPK蛋白表达量随Met浓度增加而显著增加(P<0.05).与对照组相比,PA组CCL2、ICAM1的mRNA表达量增加(P<0.05);与PA组相比,Met+PA联合组细胞CCL2和ICAM1的mRNA表达量下降(P<0.05).结论 Met可以减轻由饱和脂肪酸诱导细胞黏附因子和趋化因子表达增加而引起的大鼠H9C2型心肌细胞损伤.     

丹参单体对TNF-α诱导大鼠动脉平滑肌细胞NF-Κb和ICAM-1的影响

目的观察丹酚酸B和丹参酮ⅡA对肿瘤坏死因子(TNF-α)诱导的血管平滑肌细胞(VSMC)核因子κB(NF-κB)和细胞间黏附分子(ICAM-1)表达的影响,探讨丹参单体抗动脉粥样硬化(AS)的作用机制。方法体外培养大鼠主动脉VSMC;RT-PCR法和免疫细胞化学法检测VSMCICAM-1mRNA和ICAM-1蛋白的表达;细胞ELISA和免疫细胞化学法检测NF-κB的表达。结果TNF-α上调VSMCICAM-1mRNA和ICAM-1蛋白的表达,提高NF-κB水平(P<0.01)。两种丹参单体都抑制上述因子的表达,以丹酚酸B效果更佳。结论丹参单体抗AS的机制之一是抑制炎症相关因子的表达,丹酚酸B抗炎效果优于丹参酮ⅡA。     

氧化型低密度脂蛋白对人THP1细胞 Notch1表达及分泌细胞因子的影响

探讨氧化型低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)刺激人单核细胞株THP1对Notch1表达及分泌细胞因子的影响探讨对动脉粥样硬化(atherosclerosis,AS)发病的可能作用。方法：将人单核细胞株（THP1）经氟波酯 （P M A）刺激转化为巨噬细胞后给予不同浓度ox-LDL刺激,相差显微镜动态观察细胞形态变化,RT-PCR检测不同浓度ox-LDL刺激后细胞表面Notch1 mRNA水平,用Western-blot测定Notch1蛋白表达,用ELISA法测定上清液中血管细胞黏附分子1（vascular cell adhesive molecule1,VCAM-1）和单核细胞趋化分子1（Monocyte chemoattractant protein-1,MCP-1）浓度。结果：ox-LDL诱导48h后巨噬细胞发生树突样细胞形态改变;与对照组相比,OX-LDL能刺激巨噬细胞表面Notch1蛋白和mRNA表达升高(P<0．05),培养上清液中VCAM-1和MCP-1的表达升高(P<0．05),在50ng/ml浓度下诱导最佳。结论：ox-LDL能够刺激THP1细胞诱导Notch1表达增加,同时能增加动脉粥样硬化相关细胞因子VCAM-1和MCP-1的分泌,ox—LDL致动脉粥样硬化作用可能部分由Notch1介导。     

白细胞介素-13对肾小球系膜细胞TNF-α表达的影响

目的：研究白细胞介素—13(IL—13)对肾小球系膜细胞(MC)表达TNF—α的调节作用。方法：采用ELISA法测定MC培养上清中TNF—α。用逆转录聚合酶链式反应(RT—PCR)检测MCTNF—α mRNA表达。并用凝胶成像分析系统作半定量比较。结果：未经任何刺激的MC不分泌TNF—α，亦无TNF—αmRNA表达。经脂多糖(LPS)(10mg／L)刺激，MC高表达TNF—αmRNA及其蛋白。IL—13浓度1mg／L、10mg／L、100mg／L时均显著抑制LPS诱导MCTNF—αmRNA表达及其蛋白分泌。IL—13(100mg／L)几乎完全抑制MC表达TNF—αmRNA及其蛋白分泌，IL—13(0．1mg／L)则无抑制作用。结论：IL—13可能通过抑制MC分泌炎症细胞因子而延缓，肾内炎症过程。     

地塞米松对肾小球系膜细胞肿瘤坏死因子α产生和基因表达的影响

为观察地塞米松对内毒素诱导的肾小球系膜细胞肿瘤坏死因子α（TNFα）的产生及其mRNA表达的影响，采用免疫学和分子生物学方法，对正常、内毒素诱导后及加入地塞米松的内毒素诱导后大鼠系膜细胞的TNFα水平及TNFαmRNA表达进行了检测。结果：（1）TNFα在正常系膜细胞中分泌量极微，TNFαmRNA的表达处于低水平；（2）内毒素诱导系膜细胞TNFα的产生及mRNA表达增加；（3）地塞米松对内毒素诱导的系膜细胞TNFα的产生及其mRNA表达有明显的抑制作用，呈剂量依赖关系。结论：地塞米松是系膜产生细胞团子TNFα的较强抑制剂。     

α-硫辛酸抑制高糖诱导的系膜细胞增殖及细胞间黏附分子1的表达

目的:探讨α-硫辛酸对高糖诱导的大鼠系膜细胞增殖及细胞间黏附分子1(ICAM-1)表达的影响.方法:体外条件下采用正常糖浓度(5.6 mmol/L,NG)、高糖浓度(25 mmol/L,HG)及HG 不同浓度α-硫辛酸(50、100、200、300 μmol/L)分别与大鼠系膜细胞共同培养不同时间(12、24、48 h).MTT法测定系膜细胞增殖;RT-PCR法检测细胞ICAM-1 mRNA的表达;ELISA法测定细胞培养上清ICAM-1蛋白的浓度. 结果:50～300 μmol/L的α-硫辛酸可抑制系膜细胞增殖.高糖刺激24 h时, 200 μmol/L的α-硫辛酸干预组ICAM-1的蛋白浓度[(288.4±23.4) ng/ml]明显低于HG组[(542.3±35.6) ng/ml,P<0.01].100 μmol/L及200 μmol/L的α-硫辛酸均可下调高糖诱导的ICAM-1 mRNA的表达.结论:一定浓度的α-硫辛酸可抑制高糖诱导的系膜细胞增殖,并可降低ICAM-1蛋白和mRNA的表达.     

小剂量阿司匹林引起胃黏膜损伤机制的研究

目的：研究小剂量阿司匹林引起的胃黏膜损伤及肿瘤坏死因子TNF—α、细胞间黏附分子ICAM—1在其发病机制中的作用。方法：在既往实验的基础上，用5．021mg／kg小剂量阿司匹林灌服大鼠，观察大鼠胃黏膜损伤情况，免疫组化检测TNF—α及ICAM一1在胃黏膜细胞的表达。结果：用5．021mg／kg小剂量阿司匹林灌胃后3d出现胃黏膜损伤，14d损伤达高峰，损伤的发生率为83．3％。免疫组化结果示用小剂量阿司匹林后TNF—α3d开始表达，14d达高峰；ICAM-13d开始表达，21d达高峰。结论：小剂量阿司匹林可造成胃黏膜损伤，TNF-α及ICAM-1在小剂量阿司匹林引起的胃黏膜损伤中具有一定的作用。     

光甘草定对HUVEC细胞粘附分子ICAM-1表达影响及作用机理研究

目的观察光甘草定对人脐静脉内皮细胞(HUVEC)的细胞黏附分子1(ICAM-1)表达的影响,探讨光甘草定可能的抗动脉粥样硬化作用机理。方法体外培养HUVEC,用不同浓度的光甘草定溶液处理1h,并用肿瘤坏死因子-(TNF-)10ng/ml诱导24h。采用蛋白免疫印迹(Western Blot)、逆转录-聚合酶链反应(RT-PCR)方法检测ICAM-1 mRNA和蛋白水平的表达;用电泳迁移率变动分析(EMSA)方法检测核转录因子NF-B活性作用。结果光甘草定对TNF-诱导的HUVEC ICAM-1的蛋白表达和mRNA表达有抑制作用,呈浓度依赖性;核转录因子NF-B活性也随着光甘草定的浓度增加而降低。结论光甘草定抑制TNF-诱导的HUVEC的ICAM-1表达,可能对阻止血单核细胞向血管内皮细胞聚集和黏附、延缓动脉粥样硬化的发生和发展有一定作用。     

盆腔炎颗粒对慢性盆腔炎大鼠NF-KB及相关指标表达的影响

目的：研究盆腔炎颗粒对慢性盆腔炎大鼠核因子KB（NF-KB）及相关指标表达的影响。方法：以混合细菌加机械损伤法复制大鼠慢性盆腔炎模型,随机分为正常对照组、假手术组、模型组、盆炎净组及盆腔炎颗粒大、中、小剂量组,造模2周后,各治疗组灌胃给药2周,免疫组化法检测各组大鼠子宫组织中NF—KB、核因子KB结合蛋白（IKB）、细胞间黏附分子-1（ICAM-1）、肿瘤坏死因子-α（TNF-α）的表达,逆转录-聚合酶链反应法检测各组大鼠子宫组织中ICAM-1mRNA、TNF—αmRNA表达。结果：盆腔炎颗粒可以显著降低模型大鼠NF—KB、IKB、TNF—α、ICAM-1、TNF-αmRNA及ICAM-1mRNA的表达,盆腔炎中剂量组子宫组织NF—KB与TNF—αmRNA及ICAM-1mRNA表达之间均呈显著正相关。结论：NF—KB活化的抑制在活血补肾法治疗慢性盆腔炎抗炎及抗粘连机制中起重要作用。     

缺血预处理对缺血再灌注的大鼠肝脏中肿瘤坏死因子—α、细胞间黏附分子—1 mRNA表达的影响

目的 通过观察缺血预处理对缺血再灌注大鼠肝脏中的一些生化指标的变化,以及肿瘤坏死因子－α和细胞间粘附分子－1 mRNA表达的变化,研究缺血预处理对缺血再灌注损伤的干预方式及干预程度。方法 利用大鼠肝脏缺血再灌注损伤模型,比较缺血预处理组与缺血再灌注组以及各预处理组间在血清天冬氨酸转氨酶、丙氨酸转氨酶、乳酸脱氢酶,肝组织中丙二醛、过氧化物歧化酶等生化指标的含量变化,并观察了组织中中性粒细胞（PMNs)浸润量,研究了TNF－α及ICAM－1 mRNA表达的变化。结果 各预处理组酶的漏出和脂质过氧化物的形成减少,抗氧自由基能力增强,组织中PMNs浸润量降低。TNF－α和ICAM－1mRNA表达减少。结论 缺血预处理对肝脏缺血再灌注损伤有显著的保护作用。一次缺血10min再灌注10min的预处理保护效果最佳。缺血预处理的干预机制可能是通过下调TNF－α和ICAM－1 mRNA表达,减轻PMNs在病变部位聚集及其所产生的病理损伤作用而实现的。     

活血注射液对ox-LDL诱导的单核-内皮细胞黏附作用的影响

目的观察活血注射液对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVEC)中细胞间黏附分子-1(ICAM-1)的表达及与人单核细胞黏附作用的影响。方法以培养HUVEC作为靶细胞,在内皮细胞培养基中加入ox-LDL制备细胞损伤模型。采用蛋白定量法检测HUVEC与单核细胞的黏附率;RT-PCR检测HUVEC中ICAM-1的mRNA表达;用流式细胞仪测定HUVEC中ICAM-1的蛋白表达。结果ox-LDL作用HUVEC后12、24h时,人单核细胞与HUVEC的黏附率显著升高,HUVEC中ICAM-1的mRNA和蛋白表达水平也均明显升高,均明显高于正常对照组(P<0.01),而活血注射液可明显降低人单核细胞与HUVEC的黏附率,以及显著降低ICAM-1的mRNA和蛋白的表达水平(P<0.05,P<0.01),这种作用随着剂量的增加而增强。结论活血注射液能通过下调内皮细胞表面黏附分子的表达抑制单核-血管内皮细胞黏附,从而发挥对血管内皮细胞的保护作用,有利于减少或抑制动脉粥样硬化的形成。     

Expression of ICAM-1, B7.1 and TPO on human thyrocytes induced by IFN-alpha

Objective　To detect expression of intercellular adhesion molecule-1 (ICAM-1), B7.1 and thyroid peroxidase (TPO) on thyrocyte and study the possible mechanism of interferon-alpha (IFN-α) in the pathogenesis of autoimmune thyroid disease (AITD). Methods　Thyrocytes were cultured from 6 normal persons. Antigen expression on thyrocytes induced by cytokines was examined using immunofluorescence staining with flow cytometer. Results　IFN-α significantly stimulated the expression of ICAM-1, B7.1 and TPO, as compared with those of control group. IFN-γ markedly enhanced the expression of HLA-DR and ICAM-1, but not B7.1. Prolactin (PRL) resulted in increased expression of ICAM-1, B7.1, as well as overexpression of TPO, which is more significant than that stimulated by IFN-α. Conclusions　Thyroid autoimmunity induced by IFN-α is associated with the expression of ICAM-1, B7.1 and TPO. IFN-γ could not induce the expression of B7.1, therefore it is not an initiator in AITD. In addition, we should pay more attention to PRL which possibly plays an important role in the initiation and perpetuation of postpartum thyroiditis.     

益气活血复方含药血清对人脐静脉内皮细胞TLR4／NF—KB信号通路及TNF—a ICAM-1 mRNA表达的影响

目的：研究益气活血复方含药血清对脂多糖（LPS）诱导的人脐静脉内皮细胞（HUVECs）Toll样受体4（TLR4）／NF—KB及肿瘤坏死因子-a（TNF—a）、细胞间黏附分子（ICAM-1）mRNA表达的影响,探讨益气活血复方含药血清防治动脉粥样硬化（AS）的机制。方法：（1）选择新西兰大耳白兔20只,随机分为4组,即正常组、中药高浓度组、中药中浓度组、中药低浓度组,每组5只。以上各组白兔分别以生理盐水和高、中、低浓度益气活血复方连续灌胃7天。末次灌胃给药2h后,心脏采血,离心后分离血清。（2）体外培养人脐静脉内皮细胞,用LPS刺激后,分别加入高、中、低浓度益气活血复方含药血清干预24h,收集细胞,用荧光定量PCR方法测定TLR4、NF—KB、TNF—a及ICAM-1mRNA的表达。结果：用LPS刺激人脐静脉内皮细胞后,引起TLR4、NF-KB、TNF-a及ICAM-1 mRNA的高表达（与空白对照组比较P〈0．01）,用益气活血复方含药血清干预以后显著抑制TLR4、NF-KB、TNF-a及ICAM-1 mRNA的高表达（与模型组比较P〈0．01或P〈0．05）。结论：益气活血复方可阻断TLR4／NF—KB信号通路的高表达,同时抑制TNF—a及ICAM-1的表达,这可能是其发挥抗动脉粥样硬化作用的机制之一。     

ICAM-1在缺氧-再氧化引起的白细胞内皮细胞粘附中的作用

目的探讨细胞间粘附分子(ICAM-1是否介导缺氧-再氧化引起的中性粒细胞(PMN)和血管内皮细胞(VEC)的粘附反应.方法血管内皮细胞(VEC)经缺氧再氧化(H/R)处理后,加入PMN,用计数法检测粘附率,以细胞免疫化学及原位杂交法检测ICAM-1及ICAM-1mRNA表达.结果VEC经H/R处理后,PMN与其粘附率增高1倍(P＜0.01),ICAM-1单克隆抗体(mAb)与CD11a/CD18mAb可明显降低粘附率的增高,H/R能增加VEC的ICAM-1及ICAM-1mRNA表达.结论ICAM-1介导H/R后的PMN-VEC间粘附反应.     

Selenium inhibits high glucose- and high insulin-induced adhesion molecule expression in vascular endothelial cells

BACKGROUND: Initiation of an atherosclerotic lesion requires endothelial expression of adhesion molecules. Selenium (Se), a biologically essential trace element, can inhibit cytokine (e.g., TNF-alpha)-induced expression of adhesion molecules. Atherosclerosis is accelerated in diabetic patients. This is at least partially caused by hyperglycemia and hyperinsulinemia increasing adhesion molecule expression. These experiments tested whether Se can also alter high glucose- and high insulin-induced expression of adhesion molecules. METHODS: Human umbilical vein endothelial cells (HUVECs) were pretreated with Se and stimulated by high glucose or high insulin. Expression of adhesion molecules was measured by Western blot. RESULTS: Se (100 nmol/L) significantly inhibited glucose (25 mmol/L)-induced expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and E-selectin. Moreover, Se significantly inhibited insulin (100 nmol/L)-induced VCAM-1 and ICAM-1 expression, whereas high insulin had no inducing effect on E-selectin. Se also inhibited high glucose- and high insulin-induced activation of p38 mitogen-activated protein kinase (p38), which indicated that the preventive effects of Se on adhesion molecules may be associated with p38. The important role of p38 in Se effects was further confirmed using p38 inhibitor SB203580. CONCLUSIONS: These results suggest that Se can inhibit high glucose- and high insulin-induced expression of adhesion molecules. Such antagonism is at least partially mediated through the modulation of p38 pathway. Therefore, Se may be considered as a potential preventive intervention for diabetes-accelerated atherosclerosis.     

硫辛酸对实验性自身免疫性脑脊髓炎大鼠细胞间粘附分子-1和肿瘤坏死因子-a表达的影响

目的:探讨硫辛酸(lipoic acid LA)对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomy-elitis,EAE)大鼠的免疫抑制作用及其机制。方法:制备EAE动物模型,随机分为EAE组、LA治疗组和佐剂组。LA治疗组从免疫后(post-inoculation,免疫后)0d~5d每日腹腔内注射LA(100mg/kg)一次。在免疫后6d、8d、10d、12d、14d、16d处死动物,取脑和脊髓用免疫组化技术检测不同部位细胞间粘附分子-1(intercellular adhesion molecule-1,ICAM-1)和肿瘤坏死因子-a(tumor necrosis factor-a TNF-a)的表达。结果:LA治疗组临床症状评分(0.66±0.89)较EAE组(2.35±1.78)降低(P<0.05);LA治疗组体重减轻(7.63±3.47)较EAE组(12.60±3.53)减少(P<0.05);LA治疗组发病率(26%)较EAE组(67%)降低(P<0.05)。在免疫后10d~16d,LA治疗组ICAM-1的表达较EAE组降低(P<0.05),LA治疗组TNF-a的表达较EAE组降低(P<0.05)。结论:LA能够有效抑制EAE,其抑制机制与LA下调ICAM-1、TNF-a的表达有关。     

Effect of selenium supplementation on activity and mRNA expression of type 1 deiodinase in mice with excessive iodine intake

INTRODUCTION Previous studies[1] revealed that the relationship between the iodine intake level of a population and the occurrence of thyroid diseases is U-shaped. When insufficient dietary iodine is consumed, goiter usually develops, with occasional indu…     

Effects of rosiglitazone on the expression of beta1, integrin and ICAM-1 in high glucose-induced rat glomerular mesangial cells]

OBJECTIVE: To observe the effects of PPARgamma activators thiazolidinediones (Rosiglitazone) on the expression of intercellular adhesion molecule-1 (ICAM-1) and beta1 integrin in high glucose-induced rat glomerular mesangial cells (GMC) in order to elucidate the relationship between PPARgamma and adhesion molecules. METHODS: Rat HBZY-1 GMCs were cultured in vitro and divided into 9 groups: Normal Glucose group (N), High Glucose group (H), Mannitiol group (M), Normal and High Glucose plus 1, 5, 10 micromol/L Rosiglitazone. Every group was treated for 24 hours. The expression of ICAM-1 was measured by immunohistochemistry, the expression of beta1 integrin by indirect immunofluorescence staining and flow cytometry. RESULTS: It was found that high glucose can significantly increase the expression of ICAM-1 and beta1 integrin in GMCs, which is independent of the osmotic pressure. Rosiglitazone can inhibit the expression of ICAM-1 and beta1 integrin in normal and high glucose treated GMCs, and the inhibition is stronger in high glucose treated-group, which is in a dose-dependent manner. The expression of beta1 integrin is positively correlated with ICAM-1. CONCLUSION: Adhesion molecules involves in the pathogenesis of diabetic nephropathy (DN). PPARgamma activators-Rosiglitazone may exert effect on mesangial expansion and glomerulosclerosis in DN through the inhibition of expressions of adhesion molecules induced by high glucose.     

Perfluorocarbon attenuates lipopolysaccharide-mediated inflammatory responses of alveolar epithelial cells in vitro

Background  Toll-like receptor-4 (TLR-4) is integrally involved in lipopolysaccharide (LPS) signaling and has a requisite role in the activation of nuclear factor-κB (NF-κB). The exact mechanisms that lend perfluorocarbon (PFC) liquids a cytoprotective effect have yet to be elucidated. Therefore we examined in an in vitro model the cytoprotective effect of PFC on LPS-stimulated alveolar epithelial cellls (AECs). 

Methods  AECs (A549 cells, human lung adenocarcinoma cell line) were divided into four groups: control, PFC, LPS and LPS + PFC (coculture group) groups. Intercellular adhesion molecule-1 (ICAM-1) was detected by ELISA, tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) were detected by radioimmunological methods. The expression of TLR-4 mRNA and protein was detected by real time PCR and Western blotting, respectively. The activation of NF-κB was detected by Western blotting (proteins of I-κBα and NF-κB p65). 

Results  ICAM-1, TNF-α and IL-8 were significantly increased in LPS-stimulated AECs groups. The expression of TLR-4 mRNA and protein in LPS-stimulated groups was markedly increased. Meanwhile, NF-κB was activated as indicated by the significant degradation of IκB-α and the significant release of NF-κB P65 and its subsequent translocation into the nucleus. There were no significant effects of PFC alone on any of the factors studied while the coculture group showed significant downregulation of the secretion of ICAM-1, TNF-α and IL-8, the expression of TLR-4 mRNA and the activity of NF-κB. 

Conclusions  Taken together, our results demonstrate that LPS can induce AEC-related inflammatory injury via the activation of TLR-4 and subsequent activation of NF-κB. PFC is able to protect AECs from LPS-induced inflammatory injury by blocking the initiation of the LPS signaling pathway, which is indicated by the significant decrease of TLR-4 expression and NF-κB activation.