核因子KB在被动致敏的人气道平滑肌细胞增殖中的信号转导作用

目的　探讨核因子κB(NF κB)是否参与被动致敏的人气道平滑肌细胞 (HASMC)增殖及是否是蛋白激酶C(PKC)激活后的下游途径。方法　用 10 %哮喘患者血清被动致敏HASMC ,以10 %非哮喘者血清为对照 ,并用吡咯烷二硫氨基甲酸 (PDTC)及 12 肉豆蔻酰 13 乙酸佛波酯 (PMA)干预HASMC ,用流式细胞术、MTT法及增殖细胞核抗原 (PCNA)免疫荧光技术检测HASMC增殖 ;NF κBp6 5 免疫荧光技术及电泳迁移率改变分析 (EMSA)检测NF κB活性。结果　 (1)哮喘血清处理的HASMC ,S期细胞比例、吸光度值 (A值 )、PCNA表达阳性率、NF κBp6 5阳性率及EMSA灰度值均较对照血清组增加 (均P <0 0 5 ) ,PDTC预处理后上述指标均下降 (均P <0 0 5 )。 (2 )PMA 哮喘血清处理HASMC后 ,S期细胞比例、A值、PCNA表达阳性率、NF κBp6 5阳性率及EMSA灰度值分别为 (2 5 5 2± 3 38) %、0 5 72± 0 0 5 4、(81 2± 10 2 ) %、(2 6 5± 5 0 ) %和 716 5 4± 12 2 93,PDTC预处理后上述指标均下降 (均P <0 0 5 )。结论　NF κB参与了哮喘血清被动致敏的HASMC增殖 ,在其增殖中存在着PKC/NF κB信号途径。     

NF-κB在平滑肌细胞增殖信号转导中的调控作用

平滑肌细胞过度增殖是支气管哮喘和慢性支气管炎的气道重建、动脉粥样硬化、血管成形术后再狭窄等许多疾病的重要病因，多种刺激因素如细胞因子、生长因子、炎性介质等均能诱导平滑肌细胞增殖，但平滑肌细胞增殖的信号转导途径尚不完全清楚，研究平滑肌细胞增殖的信号转导途径对于阐明这些疾病的发病机制、选择针对性的治疗方案具有重要意义。     

蛋白激酶C-核因子κB信号转导通道对人肺动脉平滑肌细胞增殖和血管内皮生长因子表达的影响

目的　探讨蛋白激酶C(PKC) 核因子κB(NF κB)信号转导通道对人肺动脉平滑肌细胞 (HPASMCs)增殖和血管内皮生长因子 (VEGF)表达的影响。方法　体外培养HPASMCs ,用工具药PKC激活剂 12 肉豆蔻酰 13 乙酸佛波酯 (PMA)和NF κB抑制剂二硫代氨基甲酸吡咯烷 (PDTC) ,将HPASMCs分为对照组、PMA组和PMA PDTC组在常氧和缺氧条件下培养。逆转录 聚合酶链反应 (RT PCR)检测VEGFmRNA表达 ,Westernblot法检测VEGF和NF κB的抑制蛋白IκBα蛋白表达 ,免疫细胞化学法检测NF κBp6 5的表达和定位 ,流式细胞术检测细胞周期时相分布。结果　( 1)NF κBp6 5胞核染色阳性率、IκBα蛋白相对表达量及细胞周期G2 /M % :常氧或缺氧PMA组与相应对照组、PMA PDTC组比较差异均有显著性 (P均 <0 0 5 ) ;缺氧PMA组与常氧PMA组比较差异有显著性 (P <0 0 5 )。 ( 2 )VEGFmRNA和蛋白表达 :常氧对照组、PMA组、PMA PDTC组组间差异均无显著性 (P均 >0 0 5 ) ;缺氧PMA组均高于缺氧对照组、缺氧PMA PDTC组、常氧PMA组 ,差异均有显著性 (P均 <0 0 5 )。 ( 3)缺氧PMA组NF κB胞核染色阳性率、VEGF蛋白相对表达量、G2 /M %之间均呈正相关 (r =0 5 87～ 0 710 ,P均 <0 0 5 )。结论　常氧培养HPASMCs存在PKC NF κB信号转导通道 ;     

核因子κB和蛋白激酶C对哮喘Th2类细胞因子表达的调控

目的探讨核因子κB(NF-κB)和蛋白激酶C(PKC)中对支气管哮喘T淋巴细胞表达Th2类细胞因子白细胞介素4(IL-4)和IL-5的调控信号传导中的作用.方法将16只豚鼠随机分为哮喘组和正常对照组,每组8只;人体材料取自16例急性发作期哮喘患者及16名正常对照者.分别从每只豚鼠及每位受试者的外周血中分离出T淋巴细胞并分成3组培养.第1组作为空白对照,第2组加入PKC激动剂12-肉豆蔻酰-13-乙酸佛波酯(PMA),第3组同时加入PMA和NF-κB抑制剂二硫代氨基甲酸吡咯烷(PDTC).将培养的T淋巴细胞涂片,用免疫组织化学染色方法检测NF-κB的表达,用原位分子杂交方法检测IL-4和IL-5的mRNA,用酶联免疫吸附(ELISA)法检测上清液中的IL-4和IL-5.结果加入PMA培养的哮喘T淋巴细胞NF-κB活化细胞百分比、IL-4和IL-5的mRNA表达阳性细胞百分比、培养液上清中的IL-4和IL-5与空白对照组比较差异均有显著性(q=8.44～38.66,P<0.01),且与加入PMA培养的正常T淋巴细胞组比较差异也均有显著性(q=8.11～40.12,P<0.01);而同时加入PMA和PDTC培养的哮喘T淋巴细胞的以上指标与加入PMA培养的哮喘T淋巴细胞比较差异也均有显著性(q=6.50～35.63,P<0.01).T淋巴细胞NF-κB活化细胞的百分比与IL-4和IL-5的mRNA表达阳性细胞的百分比均呈显著正相关(r=0.60～0.82,P均<0.001),与培养液上清中的IL-4和IL-5也均呈显著正相关(r=0.42～0.70,P均<0.005或0.001).结论T淋巴细胞PKC活化后使IL-4和IL-5的表达增加的生物信号可能是通过激活NF-κB来传导的.T淋巴细胞PKC-NF-κB信号传导途径的激活可能是哮喘的发病机制之一.     

氨茶碱对人气道平滑肌细胞增殖抑制及NF-κB表达的影响

目的探讨氨茶碱对人气道平滑肌细胞（HASMC）生长及核因子-κB（NF-κB）表达的影响。方法原代培养HASMC,第4～10代细胞用于实验,细胞用不同浓度氨茶碱（0、50、100、200、400mg/L）诱导培养24h;流式细胞术检测HASMC凋亡和细胞周期动力学变化;免疫细胞化学染色法检测HASMC中NF-κB蛋白表达;运用实时荧光定量聚合酶链反应检测HASMC的NF-κBmRNA水平的变化。结果氨茶碱可以促进气道平滑肌细胞凋亡,而且随着氨茶碱浓度的提高,NF-κBmR-NA和蛋白的表达量均逐渐减少。结论氨茶碱可以通过降低NF-κB的表达进而抑制HASMC的增殖并促进凋亡发生。     

蛋白激酶C在支气管哮喘模型大鼠气道平滑肌细胞增殖中的信号转导机制研究

目的　探讨蛋白激酶C(PKC)信号转导途径在支气管哮喘 (简称哮喘 )大鼠气道平滑肌细胞 (ASMC)增殖中的作用。方法　 (1) 4 8只Wistar大鼠分为哮喘组 (A组 )及对照组 (B组 ) ,根据激发时间 (2、4、8周 )又分别分为A1、A2 、A3 组和B1、B2 、B3 组 ,其中A、B组大鼠各 12只 ,A2 、A3 、B2 及B3 组各 6只。用流式细胞术、四甲基偶氮唑盐 (MTT)法、增殖细胞核抗原 (PCNA)染色等方法观察每组ASMC增殖 ;(2 )用PKC激活剂 12 肉蔻酰 13 乙酸佛波酯 (PMA)及抑制剂Ro 31 82 2 0分别干预A1、B1组ASMC ,观察ASMC增殖的变化 ;(3)用逆转录 聚合酶链测定 (RT PCR)和免疫细胞化学法检测A1、A2 、A3 组和B1组ASMCPKC α的表达。结果　 (1)A组ASMCS期比例、吸光度 (A)值、PCNA表达增高 ,与B组比较差异有显著性 (P <0 0 1)。 (2 )A1组ASMCS期比例、A值、PCNA阳性表达率在干预前分别为 (19± 3) %、0 4 5 9± 0 0 36、(80± 10 ) % ;10nmol/LPMA处理后分别为 (2 7± 4 ) %、0 5 99± 0 0 78、(95± 9) % ;5 0nmol/LPMA处理后为 (14± 3) %、0 346± 0 0 38、(5 3± 8) % ;Ro 31 82 2 0处理后为 (14± 3) %、0 343± 0 0 4 8、(4 9± 8) %。各干预剂处理后与处理前比较差异均有显著性 (P <0 0 1) ;5 0nmol/LPMA处     

一氧化氮对被动致敏人气道平滑肌细胞钾通道的作用

一氧化氮（NO）是一种重要的细胞信号分子，参与多种病理生理过程。既往动物实验发现硝普钠（SNP）能通过开放钾通道松弛大鼠气道平滑肌，并且在单细胞水平上SNP（NO的供体）能使支气管哮喘（简称哮喘）大鼠模型的气道平滑肌细胞的大电导依赖性钾通道（BKCa）和电压依赖性钾通道（Kv）开放，但一氧化氮对哮喘患者的气道平滑肌细胞（HASMC）钾通道作用尚不十分清楚。我们采用全细胞膜片钳技术，通过哮喘患者血清致敏培养的人气道平滑肌细胞，观察SNP对BKCa和Kv电流及细胞兴奋性的影响。     

核因子κB信号通路在肺癌中的作用

核因子κB(nuclear factor-κB,NF-κB)是调控多种基因表达的核转录因子,其活化失调不仅与各种炎症性疾病有关,而且在肺癌的发生、发展、转移及治疗中也发挥着重要的作用.近年来研究发现,NF-κB可能是抗癌药物治疗的靶点,为肺癌等恶性肿瘤的治疗提供了新思路.     

核因子κB与支气管哮喘发病关系及其临床意义

核因子κB(nuclear factor-κB,NF-κB)是一种调控基因表达的蛋白质核转录因子.它可以调控与炎症免疫反应有关的炎症因子、细胞因子、氧化应激相关酶等的基因表达,从而参与肺部的炎症及免疫反应,在支气管哮喘(简称哮喘)的发病机制中发挥着重要作用.NF-κB抑制剂通过阻断NF-κB激活通路上的多个环节,以达到治疗哮喘的目的,将会成为哮喘治疗研究的热点.     

核因子κB与胶质细胞在阿尔茨海默病中的研究进展

近年研究发现核因子κB(nuclear factor，NF-κB)作为能调节多种基因表达的核转录因子，在神经退行性疾病中有重要作用，且NF—κB在胶质细胞中的激活及其产生的级联反应，对于阿尔茨海默病(AD)发生发展有重要影响，越来越受到人们的重视。本文就核因子κB在神经胶质细胞的作用机制及其对AD发病的影响作简要综述。     

细胞外信号调节激酶反义寡核苷酸对支气管哮喘血清被动致敏的人气道平滑肌细胞的影响

目的 观察细胞外信号调节激酶(ERK)反义寡核苷酸(ODNs)对支气管哮喘(简称哮喘)血清被动致敏的人气道平滑肌细胞(HASMCs)增殖和凋亡的影响.方法 用10%哮喘患者血清被动致敏HASMCs(空白对照组),并用脂质体将反义(AODNs组)、正义(SODNs组)及错配ERKODNs(RODNs组)导入HASMCs,以10%非哮喘患者血清为对照(对照血清组).采用流式细胞仪、四甲基偶氮唑盐(MTT)法、3H-TdR掺入法及增殖细胞核抗原(PCNA)免疫荧光技术检测HASMCs的增殖,原位末端标记法和Annexin-V FITC PI双染色法检测细胞凋亡,RT-PCR和Western blot检测ERKmRNA和ERK1/2、磷酸化ERK1/2蛋白的表达.结果 用ERK ODNs干预经哮喘血清致敏的HASMCs后,AODNs组S+G2/M期细胞所占比例、吸光度(A490)值、细胞DNA合成量和PCNA蛋白表达量[分别为(14.21±1.21)%、(0.271±0.021)、(2811±182)cpm/106和(5.25±0.60)]均低于空白对照组[分别为(22.48±2.04)%、0.507±0.090、(3869±396)cpm/106和11.25±1.21](F值分别为65.594、39.676、61.111、120.321,均P<0.01),AODNs组的凋亡指数和早期凋亡细胞百分率[分别为13.96±1.72和(9.17±0.47)%]均高于空白对照组[分别为5.37±0.05和(3.26±0.04)%],差异有统计学意义(F值分别为98.181和65.444,均P<0.01),AODNs组的ERK mRNA和ERK活化率[分别为0.43±0.06和(63±6)%]均低于空白对照组[分别为0.89±0.09和(87±8)%](F值分别为78.043和87.288,均P<0.01).而正义和错配ERK ODNs没有上述作用.结论 反义ERK可通过抑制ERK mRNA表达和翻译抑制哮喘血清被动致敏的HASMCs的增殖,促进其凋亡,ERK信号通道可能对哮喘患者HASMCs增殖与凋亡具有重要的调控作用.     

1,25-二羟维生素D3对被动致敏人气道平滑肌细胞的抑制作用

目的 检测1,25-二羟维生素D3[1,25-(OH)2D3]对被动致敏的人气道平滑肌细胞(HASMC)的增殖及其表达基质金属蛋白酶-9(MMP-9)和解整合素-金属蛋白酶33(ADAM33)的影响,探讨其调节支气管哮喘(简称哮喘)患者气道重塑的可能机制.方法用10%哮喘患者血清被动致敏HASMC,以10%非哮喘患者血清为对照.四甲基偶氮唑盐(MTT)法检测不同浓度1,25-(OH)2D3对HASMC细胞增殖活力的变化并确定其有效作用浓度.然后以有效作用浓度的1,25-(OH)2D3预处理HASMC,MTT法测定细胞增殖活力,流式细胞仪测定细胞周期,实时荧光定量PCR及蛋白免疫印迹法分别检测细胞中MMP-9及ADAM33的表达情况.结果 (1)1,25-(OH)2D3在(10-9～10-7)mol/L浓度下能浓度依赖性地抑制被动致敏的HASMC增殖(P<0.05);(2)10-7 mol/L的1,25-(OH)2D3能时间依赖性地抑制被动致敏的HASMC增殖并特异性抑制细胞周期中G1/S的转化;(3)VD组MMP-9及ADAM33蛋白表达较哮喘组分别下降了(63.4±3.6)%和(50.9±2.9)%,但仍显著高于对照组(P<0.01);(4)VD组MMP-9及ADAM33 mRNA表达较哮喘组分别下降了(52.2±2.5)%和(67.8±3.2)%,但仍显著高于对照组(P<0.01).结论 1,25-(OH)2D3能从多个层面抑制被动致敏的HASMC的功能,这可能是其调节哮喘气道重塑的作用机制之一.

Abstract：

Objective To investigate the effects of 1,25-(OH)2D3 on the proliferation of passively sensitized human airway smooth muscle cells(HASMCs) and their expressions of MMP-9 and a disintegrin and metalloprotease 33(ADAM33). Methods HASMCs were passively sensitized with 10% serum from asthmatic patients. MTT colorimetri assay was used to examine the effect of 1,25-(OH)2D3 on cell proliferation at different concentrations(10-10 mol/L, 10-9 mol/L, 10-8 mol/L, 10-7 mol/L).By this way, its optimal inhibitory concentration was determined. And then the effects of 1,25-(OH)2D3 at the optimal concentration on cell proliferation was examined by the same MTT assay and cell cycle analysis by flow cytometry. The expressions of MMP-9 and ADAM33 in HASMCs were studied by real-time quantitative RT-PCR and Western blotting analysis. Results (1)Inhibition of cell proliferation by 1,25-(OH)2D3 was barely detectable at 10-10 mol/L. But with the increasing concentration ranging from 10-9 mol/L to 10-7 mol/L, 1,25-(OH)2D3 markedly inhibited the cell proliferation concentration-dependently and reached the maximum effect at the concentration of 10-7 mol/L.Accordingly,10-7 mol/L was chosen as the optimal concentration of 1,25-(OH)2D3 for the following study. (2)At the concentration of 10-7 mol/L,1,25-(OH)2D3 inhibited the cell proliferation of passively sensitized HASMCs in a time-dependent manner and hampered the G1/S transition. (3)1,25-(OH)2D3 pretreatment attenuated the MMP-9 and ADAM33 protein levels in passively sensitized HASMCs by (63.4±3.6)% and (50.9±2.9)%,respectively (P<0.01). (4)1,25-(OH)2D3 significantly inhibited the MMP-9 and ADAM33 mRNA levels in passively sensitized HASMCs by (52.2±2.5)% and (67.8±3.2) %, respectively (P<0.01). Conclusion 1,25-(OH)2D3 has a direct inhibitory effect on passively sensitized HASMCs in vitro,including the inhibition of cell proliferation and the expressions of MMP-9 and ADAM33,which maybe associated with the beneficial role of 1,25-(OH)2D3 in the prevention and therapy of asthmatic airway remodeling.     

Effect of 1,25-(OH)2D3 (a vitamin D analogue) on passively sensitized human airway smooth muscle cells

BACKGROUND AND OBJECTIVES: In asthma, airway smooth muscle cell (ASMC) hyperplasia plays an important role in airway remodelling. Increased expression of matrix metalloproteinases-9 (MMP-9), a disintegrin and metalloprotease 33 (ADAM33) in ASMCs are also relevant to asthmatic airway remodelling. 1,25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)) has potent antiproliferative properties in vitro in various cell types; however, its role in ASMCs is not well understood. This study investigated the effect of 1,25-(OH)(2)D(3) on passively sensitized human bronchial (airway) smooth muscle cell (HASMC) proliferation and MMP-9 and ADAM33 expressions. METHODS: The effect of 1,25-(OH)(2)D(3) on cell proliferation was examined by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide colorimetry assay; cell cycle analysis by flow cytometry; and immunocytochemical staining for proliferating cell nuclear antigen (PCNA). The expression of MMP-9 and ADAM33 in HASMCs was investigated by real-time quantitative PCR and Western Blot analysis. RESULTS: 1,25-(OH)(2)D(3) effectively suppressed passively sensitized HASMC proliferation, proliferating cell nuclear antigen expression and G(1)/S transition in HASMCs passively sensitized with asthmatic serum. Further analysis showed that 1,25-(OH)(2)D(3) significantly down-regulated the expressions of protein for MMP-9 and ADAM33, as well as their mRNA levels in passively sensitized HASMCs. CONCLUSIONS: 1,25-(OH)(2)D(3) has direct inhibitory effects on passively sensitized HASMCs in vitro, including inhibition of cell proliferation and expression of MMP-9 and ADAM33, suggesting a possible beneficial role for 1,25-(OH)(2)D(3) in preventing and treating asthmatic airway remodelling.     

Factors controlling transduction signaling and proliferation of airway smooth muscle

The airway smooth muscle cell is an active participant in the inflammatory response that accompanies asthma. It can interact with the epithelium and inflammatory cells to produce cytokines and cell surface molecule upregulation. Moreover, smooth muscle cells can alter the composition of the extracellular matrix proteins via changes in the production of matrix metalloproteinases and their tissue inhibitors. These properties may contribute to the increase in the amount of airway smooth muscle that characterizes the asthmatic airway wall and the remodeling that underlies the structural changes that lead to persistent asthma.     

TGF-β effects on airway smooth muscle cell proliferation, VEGF release and signal transduction pathways

Background and objective:   Airway smooth muscle (ASM) cell hyperplasia is a key feature of airway remodelling. Mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K) are key components in signal transduction associated with cell proliferation; MAPK consists of the extracellular signal-regulated kinase (ERK), p38MAPK and c-Jun NH₂-terminal kinase (JNK). The effect of transforming growth factor (TGF)-β on the proliferation of ASM cells, the release of vascular endothelial growth factor (VEGF) by ASM cells and relevant signal transduction pathways were investigated.
Methods:   ASM cells were growth-arrested for 48 h then stimulated with platelet-derived growth factor (PDGF), TGF-β and dexamethasone. ASM cells were also treated with specific inhibitors of MAPK (PD98059), PI3K (wortmannin) and JNK (SP600125). Cell proliferation and VEGF concentrations were measured.
Results:   TGF-β neither augmented ASM cell proliferation nor showed a synergistic effect on PDGF-mediated ASM cell proliferation. Dexamethasone did not suppress ASM cell proliferation. VEGF release was augmented by TGF-β stimulation in a time-dependent manner, and was further enhanced by co-stimulation with PDGF and TGF-β. Dexamethasone suppressed VEGF release significantly. TGF-β enhanced PI3K phosphorylation, while PDGF augmented both ERK and PI3K phosphorylation. Wortmannin inhibited both TGF-β- and PDGF-stimulated VEGF release.
Conclusions:   TGF-β may facilitate airway remodelling by promoting VEGF release through the PI3K pathway, rather than via ASM cell proliferation.     

花姜酮通过抗氧化应激抑制人气道平滑肌细胞的增殖

目的 探讨花姜酮对转化生长因子β (transforming growth factor-β,TGF-β)诱导人气道平滑肌细胞(human airway smooth muscle cells,HASMCs)增殖的抑制作用及其机制.方法 用CCK-8的方法检测细胞增殖;用荧光显微镜观察2,7-二氯荧光乙酰乙酸(DCFH-DA)染色后细胞内的活性氧(reactive oxygen species,ROS);用流式细胞术检测细胞内ROS水平.结果 TGF-β1(1μg/L)作用72 h后可以明显促进HASMCs的增殖(P＜0.05);花姜酮可以呈浓度依赖性的抑制HASMCs的增殖(P＜0 05),抑制细胞内ROS的水平,以花姜酮组(30 mmol/L)的抑制作用最明显(P＜0.01).结论 花姜酮可以抑制HASMCs的增殖,其机制可能是通过抑制细胞内的ROS产生.     

辛伐他汀通过氧化应激抑制人气道平滑肌细胞增殖

目的 探讨辛伐他汀(simvastatin,Sim)对血小板活化因子(platelet activating factor,PAF)诱导的人气道平滑肌细胞(human airway smooth muscle cells,HASMCs)中活性氧(reactive oxidant stress,ROS)的生成及对细胞增殖的影响.方法 用CCK-8法检测细胞增殖,2′,7′-二氯荧光乙酰乙酸(DCFH-DA)染色,用荧光显微镜成像观察细胞荧光强弱,流式细胞术检测细胞内ROS水平.结果 PAF(10-6 mol/L,24 h)的增殖率是对照组的(1.77±0.51)倍,Sim(10-5 mol/L)(0.67±0.18)可抑制PAF促增殖作用(P＜0.01).PAF 10-6 mol/L组平均荧光强度(mean fluorescence intensity,MFI)(98.89±1.28)较阴性对照组(76.79±6.05)明显增强,PAF 10-6 mol/L + Sim 10-5 mol/L组(66.40±2.87)较PAF 10-6组减弱(P＜0.01).结论 Sim可抑制PAF诱导的HASMCs增殖,可能是通过减少ROS的生成起作用.     

Responsiveness to histamine in human sensitized airway smooth muscle.

Passive sensitization of human isolated airway smooth muscle increases contractile responses to histamine. We looked to see whether this increase was due to an alteration in the relative role of histamine H1 and H2 receptors. Human bronchial spiral strips obtained at thoracotomy were passively sensitized by incubation in serum from atopic asthmatic patients to Dermatophago?des pteronyssinus and control strips were incubated in serum from healthy non-allergic non-atopic subjects. We also studied spiral strips dissected from two spontaneously sensitized human lung specimens. Cumulative concentration-response curves (CCRC) to Ca2+ (10(-5) -3 x 10(-2) M) were constructed either in the presence of 10(-5) M histamine alone or in that of the combination histamine and the H2 antagonist cimetidine (10(-5) M). Unlike in the absence of histamine, Ca2+ CCRC in the presence of histamine alone were significantly shifted to the left in the passively sensitized tissues (mean EC50: 5.7 x 10(-4) M) compared to control ones (mean EC50: 9.3 x 10(-4) M, n = 6, P < 0.05). Addition of cimetidine to histamine did not alter the Ca2+ CCRC either in the control or in passively or spontaneously sensitized airway smooth muscle. These results suggest that (i) passive sensitization increases contractile response to Ca2+ of human bronchial smooth muscle in the presence of histamine; (ii) this increase is not due to a difference in the H1- vs H2-mediated response; and (iii) H2-mediated effects do not play a significant role in spontaneously sensitized human lung as both in the non-sensitized and passively sensitized lung.     

信号转导子和激活子3通路调控大鼠血管平滑肌细胞增殖迁移机制研究

目的研究转录信号转导子和激活子3（stat3）通路与大鼠血管平滑肌细胞（VSMCs）增殖迁移的关系,明确VSMCs增殖的信号转导过程。方法应用脂质体转染Stat3反义寡核苷酸作用于大鼠VSMCs,应用酶联免疫法（ELISA）检测Stat3水平变化及MTF法检测细胞增殖状态,蛋白免疫印迹法（Western blot）检测stat3、磷酸化stat3及其靶基因产物Cyclin D1、Bcl—XL的表达。结果转染Stat3反义寡核苷酸后,大鼠VSMCs中Stat3水平明显下降（P〈0．01）,同时其增殖水平降低,而相应空白对照组、脂质体组、转染正义寡核苷酸组变化不明显。转染Stat3反义寡核苷酸的VSMCs中stat3、p-Stat 3、Cyclin D1蛋白表达水平随作用时间延长而下降（P〈0．01）,而Bcl—xL水平无明显变化。结论癌基因stat3信号通路与大鼠VSMCs增殖高度相关,可能通过其下游靶基因Cyclin D1影响其增殖,而阻断此通路则可抑制VSMCs的增殖。