原癌基因表达在哮喘气道重塑中的作用

目的观察原癌基因表达在哮喘气道重逆中的作用。方法卵蛋白致敏豚鼠 建立哮喘模型。Dot-blot,North-ern-blot分子杂交及免疫组织化学技术观察豚鼠气道上皮,肺组织中原癌基因c-fos,c-myc,c-jun和c-sis的表达及其与气道重塑的关系。结果：正常豚鼠气道及肺组织中c-fos和c-myc mRNA无或很少表达,哮喘发作后,豚鼠气道上皮及肺组织c-fos和c-mycmRNA的表达明显增加;免疫组织化学染色显示Fos,Myc,Jun及Sis在政党豚鼠低水平表达,4种原癌基因产物表达增,国,阳性反应细胞主要分布于气道上皮细胞胞质、气道及细支气的上皮细胞胞核及浸润的炎症细胞中,病理结果显示气道粘膜及小鼠气管平滑肌周围淋巴细胞、嗜酸性粒细胞及中性粒细胞浸润,气道平滑肌显著增生,结论原部达在气道重逆过程中有重要作用。     

Ras-MAPK信号转导系统在哮喘气道重塑中的作用

目的 探讨Ras-MAPK信号转导系统在哮喘气道重塑中的作用。方法 以卵蛋白诱导并激发的豚鼠哮喘模型为研究对象，选择末次诱喘后即刻至240min间数个时间点，采用Dot-blot分子杂交法检测气道ras mRNA水平。免疫组化ABC研究气道Ras和MAPK蛋白的表达情况。结果 与正常对照组相比，哮喘组豚鼠气道上皮水肿、脱落，平滑肌层及基底膜显著增厚。诱喘后ras mRNA的表达水平明显上调，30min达高峰并维持至120min。各级气道的上皮和平滑肌细胞Ras表达量显著增加，而MAPK只在气道平滑肌层呈现高表达现象。结论 Ras-MAPK信号转导系统在哮喘气道重塑早期发挥重要作用，阻断这一途径有望延缓重塑的病理过程。     

银杏苦内酯B对哮喘豚鼠气道反应性和嗜酸性粒细胞的影响

作利用豚鼠哮喘模型观察豚鼠吸入卵蛋白（ＯＡ）或血小板激活因子（ＰＡＦ）后引起的气道反应，血浆和支气管肺泡灌洗液（ＢＡＬＦ）中溶血－ＰＡＦ（Ｌｙｓｏ－ＰＡＦ）含量和肺组织中嗜酸性粒细胞（Ｅ０）改变，及ＰＡＦ拮抗剂银杏苦内酯Ｂ（ＧＢ）的影响。结果提示致敏豚鼠ＯＡ激发和正常豚鼠ＰＡＦ激发后，与对照组相比，豚鼠血浆和ＢＡＬＦ中Ｌｙｓｏ－ＰＡＦ含量增高（Ｐ〈０．０１），肺组织中Ｅ０数增多（Ｐ〈０．０１），     

川芎嗪对哮喘大鼠肺组织c-fos表达的影响

目的：探讨川芎嗪对哮喘大鼠肺组织中C-fos表达的影响。方法：将健康成年大鼠随机分为对照组、哮喘组、药物组3组，每组6只。制作卵蛋白致敏大鼠哮喘模型。采用免疫组化法观察3组大鼠肺组织中C-fos的蛋白表达，采用逆转录-聚合酶联反应技术测定3组大鼠肺组织中C-fosmRNA的表达。结果：哮喘组C-fos在肺组织中的蛋白表达明显高于对照组（P〈0．01），应用药物川芎嗪干预后，川芎嗪组C-fos蛋白表达明显低于哮喘组（P〈0．01）。哮喘组C-fos在肺组织的mRNA表达也高于对照组（P〈0．01），应用川芎嗪药物干预后，哮喘组C-fos在肺组织的mRNA表达明显低于哮喘组（P〈0．01）。结论：川芎嗪可减少哮喘肺组织C-fos的表达从而在哮喘中发挥其抗炎和抗气道重塑的作用。     

肌酸对过敏反应豚鼠呼吸道反应的抑制作用

目的 :探讨肌酸 (creatine ,Cr)对气道反应的抑制作用。方法 :采用卵白蛋白致敏和吸入激发的方法制成豚鼠哮喘模型 (哮喘组 ) ;一部分致敏后的豚鼠激发前经口给予Cr(干预组 )。用比气道传导率 (specificairwayscon ductance ,sGaw)作为气道闭塞指标 ,气道反应强度用AUC(areaundertheresponsecurve)评价。激发后第 7h行支气管肺泡灌洗 ,对其灌洗液进行细胞计数。结果 :哮喘组和干预组在激发后均一过性出现sGaw值下降 ,AUC无明显差异。但哮喘组在致敏后 3h后再次出现sGaw值下降 ,哮喘组AUC显著地高于干预组 ;哮喘组灌洗液中的细胞总数和巨噬细胞 ,嗜酸性细胞数增高 ,而干预组显著性减少。结论 :Cr一定程度可以抑制气道炎症 ,并能抑制过敏反应豚鼠的迟发型气道反应     

二甲双胍抑制哮喘模型大鼠气道重塑的研究

目的 观察二甲双胍对哮喘气道重塑的影响及其可能作用机制.方法 28只B/N大鼠随机分为对照组、哮喘组、二甲双胍干预组和雷帕霉素干预组;制备哮喘模型,以二甲双胍、雷帕霉素进行干预;末次激发48小时后测定大鼠气道阻力及气道反应性,收集肺组织标本,采用组织病理学方法观察气道炎症细胞浸润、气道壁杯状细胞增生、气道壁纤维化和重塑...     

哮喘中外周血单个核细胞白细胞介素5表达及意义研究

该文研究了哮喘豚鼠嗜酸性粒细胞（ＥＯＳ）在外周血及支气管肺泡灌洗液（ＢＡＬＦ）中的变化,用ＲＴ－ＰＣＲ半定量测定了外周血单个核细胞（ＰＢＭＣ）及支气管组织白细胞介素５（ＩＬ－５）ｍＲＮＡ表达量。结晶显示哮喘豚鼠ＥＯＳ及ＩＬ－５ｍＲＮＡ表达量均较对照组及地塞米松干预组显著升高,ＰＢＭＣ中ＩＬ－５ｍＲＮＡ表达量与支这组织ＩＬ－５表达及外周血ＥＯＳ计数成正相关。提示哮喘豚鼠ＰＢＭＣ、ＩＬ－５ｍＲＮＡ表达     

Effect of vaisartan on the expression of angiotensin II receptors in the lung of chronic antigen exposure rats

Background Many studies have suggested that angiotensin II (Ang II) and its receptors may be involved in the development of asthma.However,the expression of angiotensin II receptors (AGTR) is not clear in the lung tissue of chronic asthmatics.This study was designed to determine the relationship between airway remodeling,dysfunction and the expression of AGTRs in a rat model of asthma.Methods Rats were sensitized with ovalbumin (OVA) for 2 weeks.Sixty minutes before an inhalation challenge,the rats challenge for 30 alternative days.Acetylcholine (Ach)-induced bronchoconstriction was measured after the final antigen challenge.White cell counts in bronchoalveolar lavage fluid (BALF) and morphological changes in the airways were then assessed.The levels of transforming growth factor-beta 1 (TGF-β1) and platelet-derived growth factor (PDGF) in BALF were detected by ELISA.The levels of AGTR1 and AGTR2 mRNA and protein in lung tissues were measured by RT-PCR and Western blotting.Results AGTR1 mRNA and protein levels in repeatedly OVA-challenged rats were significantly increased as compared with negative controls.The AGTR1 mRNA expression versus white cell counts of BALF and airway wall thickness (mainly in small airways) in lungs of chronic antigen-exposed rats were positively correlated.Valsartan decreased the level of AGTR1 in repeatedly OVA-challenged rats.However,AGTR2 mRNA and protein levels in the OVA-challenged rats and accumulation and attenuated Ach-evoked bronchoconstriction in repeatedly antigen-challenged rats.Valsartan also decreased allergen-induced structural changes in rat airway (including total airway wall thickness and smooth muscle area) and the levels of TGF-β1 and PDGF in BALF.Conclusions AGTR1 expression is potentially associated with airway remodeling and dysfunction in asthma.Ang II and AGTR1 may participate in airway inflammation and airway remodeling of chronic antigen-exposed rats.Valsartan,a AGTR1 antagonist,could inhibit AGTR1 expression and partially inhibits structural airway changes as well as airway inflammation in chronic OVA-exposed rats.     

Effect of valsartan on the expression of angiotensin II receptors in the lung of chronic antigen exposure rats

Background Many studies have suggested that angiotensin Ⅱ （Ang Ⅱ） and its receptors may be involved in the development of asthma. However, the expression of angiotensin Ⅱ receptors （AGTR） is not clear in the lung tissue of chronic asthmatics. This study was designed to determine the relationship between airway remodeling, dysfunction and the expression of AGTRs in a rat model of asthma. Methods Rats were sensitized with ovalbumin （OVA） for 2 weeks. Sixty minutes before an inhalation challenge, the rats were pretreated either with valsartan （15, 30, 50 mg.kg-1.d-1） or saline intragastrically. Then the rats received an OVA challenge for 30 alternative days. Acetylcholine （Ach）-induced bronchoconstriction was measured after the final antigen challenge. White cell counts in bronchoalveolar lavage fluid （BALF） and morphological changes in the airways were then assessed. The levels of transforming growth factor-beta 1 （TGF-β1） and platelet-derived growth factor （PDGF） in BALF were detected by ELISA. The levels of AGTR1 and AGTR2 mRNA and protein in lung tissues were measured by RT-PCR and Western blotting. Results AGTR1 mRNA and protein levels in repeatedly OVA-challenged rats were significantly increased as compared with negative controls. The AGTR1 mRNA expression versus white cell counts of BALF and airway wall thickness （mainly in small airways） in lungs of chronic antigen-exposed rats were positively correlated. Valsartan decreased the level of AGTR1 in repeatedly OVA-challenged rats. However, AGTR2 mRNA and protein levels in the OVA-challenged rats and high-dose valsartan-treated rats （50 mg.kg-1.d-1） were also increased. Valsartan significantly decreased inflammatory cell accumulation and attenuated Ach-evoked bronchoconstriction in repeatedly antigen-challenged rats. Valsartan also decreased allergen-induced structural changes in rat airway （including total airway wall thickness and smooth muscle area） and the levels of TGF-β1 and PDGF in BALE Conclusions AGTR     

白三烯受体拮抗剂对哮喘小鼠气道重塑及细胞周期蛋白D1的表达影响

目的：研究应用白三烯受体拈抗剂（孟鲁司特钠）对哮喘小鼠肺组织中细胞周期蛋白D1（Cyclin D1）的表达影响及对支气管哮喘气道重塑的作用，探讨白三烯受体拮抗剂在哮喘治疗中的作用。方法：将30只BALB／c小鼠随机分为正常对照组、哮喘组、孟鲁司特钠组3组，每组10只；卵蛋白致敏和激发建立哮喘小鼠模型；HE染色观察各组气道炎症发生及气道结构改变情况；免疫组化观察肺组织中Cyclin D1的表达及定位；RT-PCR及Western-blot测定各组小鼠肺组织中Cyclin D1的mRNA和蛋白表达变化。结果：HE染色提示哮喘组与对照组相比出现嗜酸性粒细胞浸润增多、纤毛脱失、平滑肌细胞层增厚等改变．而治疗组上述改变较哮喘组为轻；免疫组化显示Cyclin D1在哮喘小鼠气道平滑肌细胞、内皮细胞、成纤维细胞中皆有表达而在对照组中表达减弱（P〈0．05），而治疗组表达量较哮喘组低（P〈0．05）；RT-PCR及Western blot检测发现Cyclin D1在哮喘组表达较对照组为高（P〈0．01），而治疗组表达量低于哮喘组（P〈0．01）。结论：哮喘小鼠肺组织中Cyclin D1表达量较正常组为高；白三烯受体拮抗剂能够抑制Cyclin D1表达，减轻气道炎症反应、延缓气道重塑进程。     

三七花总皂苷对内皮素诱导的人主动脉血管平滑肌细胞c-myc、c-fos表达的影响

目的:观察三七花总皂苷(PNFS)对内皮素(ET-1)诱导的人主动脉血管平滑肌细胞(HASMC)癌基因c-myc、c-fos表达的影响。方法:采用ET-1诱导HASMC异常增殖,应用RT-PCR方法检测PNFS对c-myc、c-fos mRNA表达的影响。结果:PNFS能明显抑制HASMC原癌基因c-myc mRNA表达(P<0.05),而对c-fos mRNA表达影响不明显。结论:PNFS可影响癌基因c-myc mRNA表达以抑制HASMC异常增殖。     

哮喘大鼠模型肺组织平滑肌肌动蛋白-αmRNA的表达

目的　探讨平滑肌肌动蛋白 (SMA) -α m RNA的表达 ,以了解其在哮喘发病机制中的作用。方法　以卵蛋白雾化复制哮喘模型 ,在激发后 3d、1周及 2周用逆转录聚合酶链反应 (RT- PCR)检测肺部α- SMA m RNA表达 ,同时用图象分析法测量大鼠气道形态学参数。结果　大鼠哮喘激发后 2周开始出现气道平滑肌肌层增厚 ,而α-SMA m RNA在哮喘激发 1周表达升高 ,2周后更加明显 ,α- SMA m RNA表达与气道平滑肌肌层增厚具有一定相关性。结论　哮喘气道重构中既有气道平滑肌的增生肥大 ,又可能有平滑肌表型改变     

麻黄水提物对哮喘豚鼠气道炎症和气道上皮细胞STAT1信号传导影响

目的探讨麻黄水提物对哮喘豚鼠气道炎症和气道上皮细胞信号转录因子与转录活化因子1（STAT1）表达水平及其信号传导途径的调控作用。方法将36只豚鼠随机分为三组：哮喘组、麻黄煎剂组及生理盐水组（正常对照组）,每组各12只。用卵蛋白腹腔注射致敏及雾化吸入激发建立哮喘模型,给予麻黄水煎剂干预治疗。用ELISA法测定其支气管肺泡灌洗液（BALF）中白细胞介素-5（IL-5）浓度,用SABC免疫组化测定气道上皮细胞STAT1表达,同时观察BALF炎性细胞数及豚鼠肺组织病理学改变,分析其相关性。结果麻黄煎剂组豚鼠肺内炎症改变明显减轻,其BALF中白细胞总数、嗜酸性粒细胞数、IL-5浓度,明显低于哮喘组,两组比较差异有显著性,哮喘组BALF中IL-5含量与支气管上皮细胞STAT1表达均明显升高,且呈正相关关系,但麻黄煎刺组STAT1的表达与哮喘组比较差异无显著性。结论麻黄水提物对哮喘豚鼠气道炎症有明显的调控作用,其作用机制可能与抑制IL-5的表达和炎性细胞聚积有关,但麻黄不能有效抑制STAT1的表达和信号传导异常。     

天一止咳糖浆的祛痰平喘作用研究

目的 观察天一止咳糖浆的祛痰、平喘作用。方法 采用小鼠酚红法和大鼠毛细管法祛痰试验、喷雾致喘法和气管螺旋条法平喘试验，观察天一止咳糖浆不同剂量组相应作用。结果 天一止咳糖浆能明显增加小鼠的酚红排出量和大鼠的排痰量，增加豚鼠的哮喘潜伏期和显著扩张支气管平滑肌。结论 天一止咳糖浆具有显著的祛痰、平喘作用。     

镁可防治哮喘豚鼠气道重塑

目的探讨镁对哮喘豚鼠气道重塑的防治作用。方法豚鼠分为正常对照组、哮喘模型组、哮喘模型延续组、高镁组、低镁组。生化分析仪测血清镁的浓度;图像分析系统测定气道壁平滑肌厚度;取肺、心、肝、脾、肾、肾上腺、脑称重,计算出脏器重量/体重。结果哮喘模型组镁低于正常对照组（P〈0.05）,而气道壁平滑肌厚度明显高于正常对照组（P〈0.01）;哮喘模型延续组血清镁低于哮喘模型组,而气道壁平滑肌厚度明显高于哮喘模型组（P〈0.01）;高镁组血清镁高于哮喘模型延续组,而气道壁平滑肌厚度明显低于哮喘模型延续组（P〈0.01）,低镁组血清镁低于而气道壁平滑肌厚度明显高于哮喘模型延续组（P〈0.01）;哮喘模型组肺重/体重、脑重/体重明显高于正常对照组（P〈0.01）;高镁组脏器重量/体重低于哮喘模型延续组（P〈0.05）,而低镁组脏器重量/体重高于哮喘模型延续组（P〈0.05）。示豚鼠慢性哮喘时存在气道重塑和血清镁降低以及肺、脑变化,低镁可加重这些变化,高镁可缓解之。结论镁可能在哮喘气道重塑中起调节作用,镁对防治哮喘气道重塑及保护重要脏器有一定作用。     

吸入冷空气对哮喘豚鼠肺内神经激肽 A含量及基因表达的影响研究

目的研究冷空气吸入刺激对哮喘豚鼠血浆及肺组织中神经激肽A(NKA)含量及NKA mRNA表达的影响,并探讨其影响的机制.方法用卵蛋白超声雾化法制作豚鼠哮喘模型;哮喘组15例:按哮喘模型制作要求致敏及诱喘,诱喘后24 h取材.冷空气刺激组15例:致敏及诱喘同哮喘组,于诱喘前5 d、诱喘当天及取材当天(诱喘后24 h),每日吸入冷空气20 min;正常对照组:用生理盐水替代卵蛋白雾化溶液,方法同上.用酶联免疫法检测各组血浆及肺组织中NKA含量;用RT-PCR方法检测肺组织中NKA mRNA的相对含量.结果哮喘组豚鼠血浆、肺组织中NKA含量及肺组织中NKA mRNA的相对含量均明显高于正常组豚鼠,差异非常显著(均P＜0.01);冷空气刺激组哮喘豚鼠,其血浆、肺组织中NKA含量及肺组织中NKA mRNA的相对含量均明高于哮喘组豚鼠,差异显著(均P＜0.05).结论NKA参与了卵蛋白诱发的豚鼠哮喘发病机制;冷空气吸入刺激能显著增加哮喘豚鼠血浆及肺组织中NKA含量;冷空气吸入刺激具有在转录水平上调NKA mRNA表达的作用,由此导致血浆及肺组织中NKA蛋白含量增多.     

THE EFFECTS OF RETINOIC ACID ON EXPRESSION OF C-MYC, C-FOS IN LEUKEMIC PROMYELOCYTES

The expression of c-myc, c-fos of leukemic promyelocytes (HL-60 and acute promyelocytic leukemia cells) from 18 acute promyelocytic leukemia (APL) patients treated with all-trans retinoic acid (RA) in vitro was studied. There was no expression of c-fos in HL-60 cells and APL cells from 17 patients. But in one case, a slight expression of c-fos in leukemic cells was observed, and the alteration of expression level was found during the treatment of the cells with RA in vitro. The expression of c-myc in HL-60 cells induced by RA was altered, decrease in the early, increase in the middle, and decline in the later stage were found. The c-myc expression in leukemic cells of eighteen APL patients was variable. There was c-myc expression in eleven APL cells, but no expression in the others. The APL cells with c-myc expression were treated with RA in vitro to observe the kinetic changes of c-myc RNA level. The results showed that the expression of c-myc was gradually decreased except in few cases. Using in situ hybridization technique for detecting the alteration of c-myc expression in leukemic cells of two APL patients. the high level of c-myc before RA treatment and low level of c-myc expression after obtaining complete remission induced by RA were found. The possibility of different proto-oncogenes implicated differentiation was discussed.     

祛风宣痹方对哮喘豚鼠细胞粘附分子的影响

[目的]探讨祛风宣痹方(平哮合剂)对支气管哮喘的作用机理.[方法]用卵蛋白致敏豚鼠复制支气管哮喘动物模型,用祛风宣痹方(平哮合剂)治疗,观察动物血浆中细胞粘附分子(ICAM-1)水平的变化.[结果]实验性哮喘豚鼠发作时血浆中细胞粘附分子(ICAM-1)水平显著高于正常对照组,祛风宣痹方(平哮合剂)对实验性哮喘豚鼠中细胞粘附分子(ICAM-1)水平有明显降低作用(95%可信区间为20.387～53.834u/ml)(P＜0.01).[结论]祛风宣痹方(平哮合剂)有效治疗哮喘的作用与抑制细胞粘附分子(ICAM-1)产生密切相关.     

哮喘豚鼠气道内NK-1R mRNA表达及其含量的研究

目的:研究哮喘豚鼠气道内神经激肽1受体(NK-1R)mRNA的分布及相对含量.方法:用卵蛋白超声雾化法制作哮喘豚鼠模型;原位杂交技术检测NK-1R mRNA,光镜观察其在气道内表达的部位.图像分析测定NK-1R mRNA的相对含量.结果:哮喘组NK-1R mRNA表达程度明显高于正常组;NK-1RmRNA分布于气管上皮细胞上层细胞表面、黏膜下层、血管周围上皮细胞、炎性细胞的表面和平滑肌细胞、腺体细胞表面.结论:NK-1R参与了卵蛋白诱导的哮喘豚鼠发病机制.     

Inhibitory effects of sunitinib on ovalbumin-induced chronic experimental asthma in mice

Background Tyrosine kinase signaling cascades play a critical role in the pathogenesis of allergic airway inflammation. Sunitinib, a multitargeted receptor tyrosine kinase inhibitor, has been reported to exert potent immunoregulatory, anti-inflammatory and anti-fibrosis effects. We investigated whether sunitinib could suppress the progression of airway inflammation, airway hyperresponsiveness （AHR）, and airway remodeling in a murine model of chronic asthma.
Methods Ovalbumin （OVA）-sensitized mice were chronically challenged with aerosolized OVA for 8 weeks. Some mice were intragastrically administered with sunitinib （40 mg/kg） daily during the period of OVA challenge. Twelve hours after the last OVA challenge, mice were evaluated for the development of airway inflammation, AHR and airway remodeling. The levels of total serum immunoglobulin E （IgE） and Th2 cytokines （interleukin （IL）-4 and IL-13） in bronchoalveolar lavage fluid （BALF） were measured by ELISA. The expression of phosphorylated c-kit protein in the lungs was detected by immunoprecipitation/Western blotting （IP/WB） analysis.
Results Sunitinib significantly inhibited eosinophilic airway inflammation, persistent AHR and airway remodeling in chronic experimental asthma. It reduced levels of total serum IgE and BALF Th2 cytokines and also lowered the expression of phosphorylated c-kit protein in remodelled airways.
Conclusions Sunitinib may inhibit the development of airway inflammation, AHR and airway remodeling. It is potentially beneficial to the prevention or treatment of asthma.