美洛昔康抑制正常人中性粒细胞与滑膜细胞粘附的作用机理研究

目的研究美洛昔康对正常人中性粒细胞(polymorphonuclear leukocyte,PMN)与滑膜细胞(human synovial cell,HSC)粘附的抑制作用及其作用机理。方法用MTT比色法检测PMN与HSC的粘附,分别用Cell-ELISA和RT-PCR法检测粘附分子ICAM-1和VCAM-1的蛋白及基因表达,用EMSA法检测NF-κB的活性。结果美洛昔康可显著的并以剂量依赖的方式抑制TNF-α(50 u·mL^-1)和IL-1β(50 u·mL^-1)作用12 h诱导的PMN与HSC粘附,其IC₅₀分别为3.38×10^-7和3.56×10^-6 mol·L^-1。进一步研究发现美洛昔康在1×10^-6～1×10^-5 mol·L^-1时还可在蛋白水平及mRNA水平抑制TNF-α(50 u·mL^-1)诱导的HSC细胞ICAM-1的表达,但对VCAM-1蛋白及mRNA表达均未见显著影响;同时美洛昔康还可显著抑制50 u·mL^-1 TNF-α诱导的NF-κB的活化。结论美洛昔康抑制NF-κB的活化,进而抑制ICAM-1的表达可能是其抑制PMN与HSC粘附的机制之一。     

Sanggenon C对人多形核白细胞和滑膜细胞粘附的抑制作用及其机制

目的:观察化合物Sanggenon C对人外周血多形核白细胞(PMN)与人滑膜细胞(HSC)粘附的抑制作用,并探讨其作用机制.方法:MTT比色法研究PMN与HSC粘附,Cell-ELISA及RT-PCR法研究HSC粘附分子ICAM-1和VCAM-1表达,EMSA研究核转录因子NF-κB的活化.结果:Sanggenon C在0.01-10μmol·L~(-1)范围内均可显著抑制TNF-α 5O kU·L~(-1)与IL-1β诱导的HSC与PMN粘附,其IC_(50)分别为27.29nmol. L~(-1)和54.45nmol·L~(-1);Sanggenon C可显著抑制HSC表面ICAM-1和VCAM-1蛋白表达,同时也显著抑制VCAM-1 mRNA表达,但对ICAM-1 mRNA表达无显著影响;Sanggenon C在1-10μmol·L~(-1)浓度下也可显著抑制TNF-α对NF-κB的活化.结论:Sanggenon C是一个有效的人PMN与HSC粘附抑制剂,其作用机制可能是通过抑制NF-κB的活化,进而抑制HSC表面VCAM-1的表达或抑制ICAM-1转录后调控过程而实现的.     

银杏内酯B对脂多糖刺激的小鼠腹腔巨噬细胞TNFα生成及大鼠胸腔多形核白细胞NF-κB活化的影响

目的研究银杏内酯B对脂多糖刺激的小鼠腹腔巨噬细胞TNFα生成及大鼠胸腔多形核白细胞NF-κB活化的影响。方法用L929细胞结晶紫染色法检测TNFα的含量，用电泳迁移率改变检测法检测NF-κB的结合活性。结果1和10 μmol·L^-1银杏内酯B能够显著抑制LPS刺激的小鼠腹腔巨噬细胞TNFα的生成，其IC₅₀为0.26 μmol·L^-1；1 mg·L^-1 LPS和1 nmol·L^-1 PAF均可活化大鼠胸腔多形核白细胞NF-κB；银杏内酯B能够抑制LPS或 PAF刺激的NF-κB活化。结论银杏内酯B能够抑制LPS刺激的小鼠腹腔巨噬细胞TNFα生成及大鼠胸腔多形核白细胞NF-κB的活化。PAF参与LPS激活NF-κB的过程。     

肺热普清散对斑马鱼炎症模型的抗炎作用及机制研究

目的 探讨藏药肺热普清散对斑马鱼炎症模型的作用及机制。方法 挑选受精后 72 h 斑马鱼随机移入 24 孔板中,分为对照组、模型组和肺热普清散低、中、高质量浓度(3、10、30 mg·L^-1)组,对照组与模型组不加药,肺热普清散与斑马鱼共孵育 1 h 后,除对照组外,其余各组用 20 μmol·L^-1硫酸铜处理 2 h 制备炎症模型。在荧光显微镜下观察绿色荧光标记中性粒细胞的 Tg(Lyz:EGFP)系斑马鱼体内炎症细胞迁移情况;实时荧光定量 PCR(qRT-PCR)检测野生型斑马鱼核因子κB2(NF-κB2)、白细胞介素(IL)-1b、肿瘤坏死因子(TNF)-α、IL-8 mRNA 水平;Western blotting 实验检测野生型斑马鱼 NF-κB、TNF-α 蛋白表达水平;免疫荧光法检测各组鱼尾 NF-κB 表达。结果 对照组斑马鱼荧光细胞主要分布于头部和主动脉部,躯干部位主动脉以上荧光细胞稀少;模型组荧光细胞在各区域均增加,显示出炎症细胞向血管外迁移的明显趋势,躯干侧线以上荧光细胞计数较对照组显著增加(P<0.001);肺热普清散 10、30 mg·L^-1组躯干部荧光细胞计数与模型组比较显著降低(P<0.01、0.001)。与模型组比较,肺热普清散 3、10、30 mg·L^-1 组 IL-1b、TNF-α、IL-8 的 mRNA 水平显著下调(P<0.05、0.01),10、30 mg·L^-1 组 NF-κB2 mRNA 水平显著下调(P<0.01);10、30 mg·L^-1 组 NF-κB、TNF-α 蛋白表达水平显著降低(P<0.01、0.001)。免疫荧光结果显示,模型组躯干部位和鱼鳍 NF-κB 白色荧光点较对照组明显增加,经 10 mg·L^-1肺热普清散处理后的相同部位荧光点减少。结论 肺热普清散对硫酸铜诱导的斑马鱼炎症模型具有抗炎作用,机制与抑制 NF-κB、TNF-α 表达有关。     

甲氨蝶呤对脂多糖诱导的大鼠脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路的影响

目的 观察甲氨蝶呤对脂多糖（lipopolysaccharide,LPS）诱导的大鼠脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路的影响。方法 脊髓组织块法培养神经胶质细胞。将分离的神经胶质细胞接种于多孔板培养48 h后,分为空白对照组、LPS组、LPS+pIκBα抑制剂组、LPS+甲氨喋呤组。随后应用免疫印迹法测定各组分的pIκBα与胞核及胞浆NF-κBp65水平变化,酶免疫法（ELISA）测定炎性因子TNF-α、IL^-1β、IL-6含量。结果 神经胶质细胞经LPS诱导后,pIκBα、胞核NF-κBp65和细胞上清液炎性因子TNF-α、IL^-1β、IL-6水平均显著增加（P<0.05或P<0.01）。甲氨喋呤可明显抑制经LPS诱导的神经胶质细胞pIκBα水平,显著降低胞核NF-κBp65水平和细胞上清液炎性因子TNF-α、IL^-1β、IL-6的含量（P<0.05）。结论 甲氨喋呤对LPS诱导的脊髓神经胶质细胞pIκBα-NF-κBp65-炎性因子通路有显著的抑制作用。     

丹参酮ⅡA对TNF-α诱导的ECV304细胞NF-κB、IκB-α表达及粘附分子ICAM-1、VCAM-1mRNA表达的影响

目的探讨丹参酮ⅡA对血管内皮细胞株ECV304NF-κB、IκB-α及粘附分子ICAM-1、VCAM-1mRNA表达的影响,以阐明其抗动脉粥样硬化作用及机制。方法通过建立TNF-α诱导的ECV-304细胞损伤模型,以抗氧化剂吡咯烷二硫代氨基甲酸盐(PDTC)做为对照,间接细胞ELISA方法定量检测NF-κB、IκB-α的表达,RT-PCR方法检测各组细胞ICAM-1、VCAM-1mRNA的表达。结果TNF-α可增加ECV304细胞转录因子NF-κB的表达,同时降低其抑制因子IκB-α的表达,低浓度的丹参酮ⅡA对TNF-α引起的ECV304细胞NF-κB的表达升高无明显抑制作用,但可增加其IκB-α的表达;高浓度的丹参酮ⅡA可明显抑制NF-κB的表达,同时增加其抑制因子IκB-α的表达。同时丹参酮ⅡA抑制TNF-α诱导的ECV304细胞ICAM-1、VCAM-1mRNA表达。结论丹参酮ⅡA可通过抑制转录因子NF-κB的激活及其相关粘附分子ICAM-1、VCAM-1mRNA表达,这有利于抑制动脉粥样硬化过程中的炎症从而发挥抗动脉粥样硬化作用。     

基于NF-κB信号通路研究前列闭尔通栓对大鼠慢性非细菌性前列腺炎的影响

目的 探讨前列闭尔通栓对自身免疫性前列腺炎（EAP）大鼠的影响及作用机制。方法 50只SD大鼠采用前列腺蛋白提纯液联合完全弗氏佐剂制备EAP大鼠模型，另取10只作为对照组，造模成功大鼠随机分为模型组、前列通栓（0.42 g·只^-1）组和前列闭尔通栓低、中、高剂量（0.33、0.66、0.99 g·只^-1）组，直肠给药28 d。压力换能器测定膀胱内压变化速率，显微镜计数测定前列腺液中卵磷脂小体、白细胞数量，ELISA法检测前列腺组织炎症因子，HE染色观察前列腺组织病理变化，Westernblotting检测前列腺组织核因子κB（NF-κB）p65、磷酸化κB抑制因子激酶（p-IKK-α）/IKK-α、肿瘤坏死因子-α（TNF-α）、磷酸化IκB激酶-α（p-IκB-α）/IκB-α、环氧化酶-2（COX-2）蛋白表达；实时荧光定量PCR（qRT-PCR）法检测重组人趋化因子配体5（CXCL5）、白细胞介素-6（IL-6）、TNF-α、COX-2基因表达。结果 与对照组比较，模型组大鼠膀胱内压变化速率显著降低（P<0.01）；白细胞数量显著增加、卵磷脂小体数量显著减少（P<0.01）；前列腺组织TNF-α、IL-8水平显著升高（P<0.01），IL-10水平显著降低（P<0.01）；前列腺组织炎症反应明显，病理评分显著升高（P<0.01）；前列腺组织NF-κB p65、p-IKK-α、p-IκB-α、TNF-α、COX-2蛋白表达显著升高（P<0.05、0.01）；前列腺组织CXCL5、COX-2、TNF-α基因表达升高（P<0.05）。与模型组比较，前列闭尔通栓中剂量组膀胱内压变化速率显著升高（P<0.01）；各剂量组白细胞数量显著减少、卵磷脂小体数量显著增加（P<0.01）；中、高剂量组TNF-α、IL-8水平显著降低（P<0.05、0.01）；各剂量组前列腺组织炎症反应明显减轻，病理评分显著降低（P<0.01）；各剂量组前列腺组织p-IκBα/IκBα、COX-2蛋白表达显著降低（P<0.01）；低剂量组前列腺组织p-IKK-α/IKK-α蛋白表达显著降低（P<0.05）；低、高剂量组前列腺组织NF-κB p65蛋白表达显著降低（P<0.05）；中剂量组前列腺组织TNF-α蛋白表达显著降低（P<0.05） ；各剂量组前列腺组织CXCL5、IL-6、COX-2基因表达显著降低（P<0.05）。结论 前列闭尔通栓可有效改善EAP大鼠前列腺组织病理形态，减轻炎症反应，其作用机制可能与抑制NF-κB信号通路相关蛋白NF-κB p65、p-IKK-α、TNF-α、p-IκB-α表达相关。     

基于网络药理学和实验验证探讨升降散治疗急性肺损伤作用及机制

目的 基于网络药理学和动物实验探究升降散治疗急性肺损伤（ALI）的作用机制。方法 在中药系统药理学分析平台数据库（TCMSP）和Swiss Target Prediction数据库中检索升降散的成分及靶点,在基因数据库（Gene Cards）中检索ALI的疾病靶点。将药物与疾病交集靶点上传至STRING数据平台进行蛋白质-蛋白质相互作用（PPI）分析,运用DAVID网站进行基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）通路分析。体内实验,除对照组外,升降散各给药组分别ig给药20 mL·kg^-1药液（散剂、水煎液高、低剂量分别为175、350 mg·kg^-1）,每天早晚各1次,共连续给药14 d;地塞米松组第12天开始ip地塞米松（5 mg·kg^-1）,连续给药3 d;除对照组外,其余各组第15天分别向气管内滴注1 mg·mL^-1脂多糖（5 mg·kg^-1）进行造模,观察肺组织病理损伤情况,ELISA法检测白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）水平,实时荧光半定量聚合酶链式反应（qRT-PCR）法检测Toll样受体4 （TLR4）、磷酸化-丝裂原活化蛋白激酶p38 （p38-MAPK）、细胞外调节蛋白激酶（ERK）、c-Jun氨基末端激酶（JNK-1） mRNA表达量,Western blotting法检测核因子κB（NF-κB）和人核因子κB抑制蛋白α（IκBα）表达量。结果 网络药理学预测出升降散活性成分32个,关键靶点292个,ALI疾病靶点有1 454个,两者交集靶点95个;升降散治疗ALI的关键靶点主要参与NF-κB、MAPK等信号通路。通过ALI小鼠模型实验结果显示,升降散能够显著减轻ALI小鼠肺部组织炎症,改善肺泡间隔及肺泡壁增厚程度;降低血清中炎症因子IL-6、IL-1β、TNF-α的水平（P＜0.05、0.01）;下调TLR4、p38-MAPK、ERK、JNK-1 mRNA的表达量（P＜0.05、0.01）;Western blotting结果表明升降散可通过抑制NF-κB蛋白磷酸化水平（P＜0.01）,激活IκBα蛋白（P＜0.01）,缓解ALI。结论 升降散能够基于多成分、多靶点、多通路的特点相互协同治疗ALI,其机制可能与NF-κB、MAPK等信号通路有关,且散剂治疗ALI的作用优于水煎液。     

异丹叶大黄素对人滑膜细胞白细胞介素-8生成及mRNA表达的影响

目的研究异丹叶大黄素(isorhapotigenin,Iso)对肿瘤坏死因子α(TNF-α)诱导的人滑膜细胞(humansynovialcell,HSC)白细胞介素-8(IL-8)生成和mRNA表达的影响.方法用RIA方法测定IL-8的含量,以RT-PCR法测IL-8mRNA.结果Iso在1×10^-6-1×10^-5mol·L^-1浓度范围内对TNF-α诱导的人滑膜细胞IL-8生成有抑制作用,并抑制TNF-α诱导的HSCIL-8mRNA表达.结论Iso抑制TNF-α诱导的HSCIL-8生成,可能与影响IL-8mRNA表达有关.     

红景天苷对乙醛刺激的大鼠肝星状细胞增殖及胶原基因表达的影响

目的探讨红景天苷对乙醛刺激的大鼠肝星状细胞(hepatic stellate cell,HSC)增殖、α1(I)型胶原mRNA合成、IκB和NF-κB表达的影响。方法用原位杂交、免疫细胞化学和凝胶电泳迁移率技术(EMSA)检测HSC的增殖和胶原基因表达。结果乙醛刺激HSC增殖、α1(I)型胶原mRNA合成 ,使IκB表达下降、NF-κB活性增加。红景天苷(0.5～2.5 mg·mL^-1)对上述影响有抑制作用,且以1.5 mg·mL^-1组效果最好。结论红景天苷明显抑制乙醛刺激的HSC增殖及胶原基因的表达。     

大鼠中性粒细胞与滑膜细胞粘附模型的建立

目的：建立大鼠中性粒细胞（PMN）与滑膜细胞（RSC）的粘附模型。方法：MTT比色法。结果：500 U.mL^-1 rhTNF-α与IL-1刺激RSC 12 h对PMN有明显的粘附作用, MTT-PMN细胞数与540 nm吸光度值有良好的线形相关性;氢化考的松（hydrocortisone, Hy）和美洛昔康（meloxicam, Melo）均可抑制PMN与500 U.mL^-1 rhTNF-α/或IL^-1诱导的RSC粘附,提示Hy和Melo治疗类风湿性关节炎（RA）可能与其抑制PMN与RSC粘附作用有关。结论：以MTT比色法为基础建立的检测大鼠PMN与RSC的粘附模型,可用于PMN与RSC粘附抑制剂的筛选。     

Hesperidin, hesperidin methyl chalone and phellopterin from Poncirus trifoliata (Rutaceae) differentially regulate the expression of adhesion molecules in tumor necrosis factor-alpha-stimulated human umbilical vein endothelial cells

The fruits of Poncirus trifoliata (L.) are widely used in Oriental medicine to treat allergic inflammation. Recently, several active compounds including hesperidin, hesperidin methyl chalone and phellopterin from P. trifoliata (Rutaceae) were isolated and characterized. The goal of this study was to investigate the differential effect of hesperidin, hesperidin methyl chalone and phellopterin derived from P. trifoliata (Rutaceae) on the induction of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by TNF-alpha and the possible molecular mechanisms by which they differentially regulate ICAM-1 and VCAM-1 expressions. Stimulation of human umbilical vein endothelial cells (HUVECs) with TNF-alpha resulted in the increase of ICAM-1 and VCAM-1 expressions, while pretreatment with the three components completely inhibited VCAM-1 expression in a dose-dependent manner but had no effect on ICAM-1 expression. All three compounds failed to block TNF-alpha-induced phosphorylation of ERK1/2, which is involved in regulating ICAM-1 production by TNF-alpha. Furthermore, they efficiently inhibited the phosphorylation of Akt and PKC, suggesting that Akt or PKC pathways are an important target by which these compounds regulate TNF-alpha-induced VCAM-1 but not ICAM-1. Additionally, treatment with these chemicals also inhibited U937 monocyte adhesion to HUVECs stimulated with TNF-alpha. Interestingly, the inhibitory effect of hesperidin, hesperidin methyl chalone and phellopterin on monocyte adhesion to HUVECs was recapitulated by transfecting cells with VCAM-1 siRNA. Taken together, hesperidin, hesperidin methyl chalone and phellopterin reduce TNF-alpha-induced VCAM-1 expression through regulation of the Akt and PKC pathway, which contributes to inhibit the adhesion of monocytes to endothelium.     

Manassantin a and b isolated fromSaururus chinensis inhibit TNF-α-induced Cell adhesion molecule expression of human umbilical vein endothelial cells

Leukocyte adhesion to the vascular endothelium is a critical initiating step in inflammation and atherosclerosis. We have herein studied the effect of manassantin A (1) and B (2), dineolignans, on interaction of THP-1 monocytic cells and human umbilical vein endothelial cells (HUVEC) and expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin in HUVEC. When HUVEC were pretreated with 1 and 2 followed by stimulation with TNF-alpha, adhesion of THP-1 cells to HUVEC decreased in dose-dependent manner with IC50 values of 5 ng/mL and 7 ng/mL, respectively, without cytotoxicity. Also, 1 and 2 inhibited TNF-alpha-induced up-regulation of ICAM-1, VCAM-1 and E-selectin. The present findings suggest that 1 and 2 prevent monocyte adhesion to HUVEC through the inhibition of ICAM-1, VCAM-1 and E-selectin expression stimulated by TNF-alpha, and may imply their usefulness for the prevention of atherosclerosis relevant to endothelial activation.     

重组人胸腺素α原体外对IFN-γ,IFN-α及TNF-α的影响

目的体外观察重组人胸腺素α原(prothymosin α,Pro Tα)对几种重要细胞因子分泌的影响。方法用脾淋巴细胞、脾巨噬细胞及腹腔巨噬细胞,以ELISA法检测Pro Tα对IFN-γ,IFN-α和TNF-α分泌的影响。结果1×10^-7 mol·L^-1 Pro Tα明显促进脾细胞分泌IFN-γ(P<0.05),该浓度的Pro Tα也明显刺激小鼠脾巨噬细胞分泌IFN-α(P<0.01);在小鼠腹腔巨噬细胞中,Pro Tα能明显刺激IFN-α和TNF-α的分泌(P<0.01)。结论Pro Tα对细胞因子IFN-γ,IFN-α和TNF-α的分泌均有促进作用。     

吲哚美辛和美洛昔康对脂多糖诱导的小鼠腹腔巨噬细胞NF-κB的抑制作用

目的　研究非甾体抗炎药吲哚美辛(indomethacin)和美洛昔康(meloxicam)对细菌脂多糖(LPS)诱导表达的C57小鼠腹腔巨噬细胞核转录因子NF-κappaB(NF-κB)的抑制作用。方法　NF-κB的测定采用电泳迁移率改变法。结果　小鼠腹腔巨噬细胞经LPS诱导后细胞NF-κB含量明显增高。Indomethacin和meloxicam在10^-5-10^-7mol.L^-1浓度下可明显降低LPS激活的小鼠腹腔巨噬细胞NF-κB活化。结论　Indomethacin和meloxicam对NF-κB活化的抑制作用可能是两者的抗炎作用机理之一。     

Magnolol attenuates VCAM-1 expression in vitro in TNF-alpha-treated human aortic endothelial cells and in vivo in the aorta of cholesterol-fed rabbits.

1. In a previous study, we showed that magnolol, a potent antioxidant derived from a Chinese herb, attenuates monocyte chemotactic protein-1 (MCP-1) expression and intimal hyperplasia in the balloon-injured aorta of cholesterol-fed rabbits. Expression of cell adhesion molecules by the arterial endothelium and the attachment of leukocytes to the endothelium may play a major role in atherosclerosis. In the present study, the effects of magnolol on the expression of endothelial-leukocyte adhesion molecules and the activation of nuclear factor kappa B (NF-kappa B) in tumour necrosis factor-alpha (TNF-alpha)-treated human aortic endothelial cells (HAECs) were investigated. 2. Pretreatment of HAECs with magnolol (5 microM) significantly suppressed the TNF-alpha-induced expression of vascular cell adhesion molecule-1 (VCAM-1) (64.8+/-1.9%), but had no effect on the expression of intercellular cell adhesion molecule-1 and endothelial cell selectin. 3. Magnolol (5 and 10 microM) significantly reduced the binding of the human monocytic cell line, U937, to TNF-alpha-stimulated HAECs (58.4 and 56.4% inhibition, respectively). Gel shift assays using the (32)P-labelled NF-kappa B consensus sequence as probe showed that magnolol pretreatment reduced the density of the shifted bands seen after TNF-alpha-induced activation. Immunoblot analysis and immunofluorescence staining of nuclear extracts demonstrated a 58% reduction in the amount of NF-kappa B p65 in the nuclei in magnolol-treated HAECs. Magnolol also attenuated intracellular H(2)O(2) generation in both control and TNF-alpha treated HAECs. 4. Furthermore, in vivo, magnolol attenuates the intimal thickening and TNF-alpha and VCAM-1 protein expression seen in the thoracic aortas of cholesterol-fed rabbits. 5. Taken together, these data demonstrate that magnolol inhibits TNF-alpha-induced nuclear translocation of NF-kappa B p65 and thereby suppresses expression of VCAM-1, resulting in reduced adhesion of leukocytes. These results suggest that magnolol has anti-inflammatory properties and may play important roles in the prevention of atherosclerosis and inflammatory responses in vivo.     

A pilot study of estrogen's effects on bronchial myocyte adhesion molecule expression

We examined the effects of estrogen on tumor necrosis factor alpha (TNF-alpha)-induced expression of intracellular adhesion molecule (ICAM-1) and vascular adhesion molecule (VCAM-1) in cultured human bronchial smooth muscle cells (BSMC). Experiments were performed in triplicate in T-75 tissue culture flasks containing normal human BSMC. Four experiments were carried out: untreated BSMC cells (control); TNF-alpha 1000 U/ml stimulation of BSMC; forskolin 5 microM before TNF-alpha stimulation of BSMC; and estradiol 30 microM before TNF-alpha stimulation of BSMC. Cyclic adenosine monophosphate was measured by a commercially available radioimmunoassay kit. Cell expression of ICAM-1 and VCAM-1 was quantified by flow cytometry Incubation of cells with TNF-alpha 1000 U/ml for 24 hours elicited a 27-fold increase in basal expression of ICAM-1 and a 2-fold increase in VCAM-1 (p>0.05). Incubation of BSMC with forskolin 5 microM, for 1 hour before TNF-alpha, decreased TNF-alpha-induced expression of ICAM-1 by 62% and VCAM-1 slightly by 17%. The BSMC incubated with estradiol 30 microM, 1 hour before TNF-alpha, decreased TNF-alpha-induced expression of ICAM-1 by 21%; VCAM-1 remained unchanged (p>0.05). We found a trend toward inhibition of TNF-alpha-stimulated ICAM-1 expression in cultured BSMC with pretreatment with estradiol. However, due to large variability within the cell culture model, statistical significance was not reached.