本文選題：支氣管哮喘　切入點(diǎn)：糖皮質(zhì)激素　出處：《北京協(xié)和醫(yī)學(xué)院》2012年博士論文

【摘要】：目的建立煙霧暴露的哮喘大鼠模型及尼古丁誘導(dǎo)人肺腺癌A549細(xì)胞糖皮質(zhì)激素抵抗的實(shí)驗(yàn)?zāi)Ｐ?觀察p38絲裂原活化蛋白激酶(p38MAPK)抑制劑SB203580對煙霧暴露的哮喘大鼠的影響,并分別從體內(nèi)實(shí)驗(yàn)、體外實(shí)驗(yàn)兩個方面探討SB203580影響糖皮質(zhì)激素敏感性的機(jī)制。 方法Wistar大鼠隨機(jī)分為4組,即正常對照組、哮喘組、煙霧暴露的哮喘組及SB203580干預(yù)組。動物肺功能儀測定大鼠呼氣阻力、吸氣阻力及肺順應(yīng)性,觀察肺組織病理學(xué)改變,通過ELISA檢測大鼠肺組織勻漿中IL-4、IL-5和IL-8的水平。經(jīng)RT-PCR檢測大鼠肺組織糖皮質(zhì)激素受體(GR)、HSP90和p38MAPK mRNA的表達(dá),經(jīng)Western blot檢測大鼠肺組織GR、HSP90和p38MAPK蛋白的表達(dá)。培養(yǎng)A549細(xì)胞,將其隨機(jī)分為四組：A組：空白對照組,B組：1umol/L的地塞米松,C組：1umol/L的地塞米松+1umol/L的尼古丁,D組：1umol/L的地塞米松+1umol/L的尼古丁+1umol/L的SB203580,采用免疫熒光染色技術(shù)分析GR亞細(xì)胞定位。 結(jié)果與煙霧暴露的哮喘組相比,SB203580干預(yù)組大鼠的氣道阻力顯著下降,肺順應(yīng)性顯著升高,差異有統(tǒng)計學(xué)意義(P0.05)；氣道炎癥明顯減輕；肺組織中IL-4、IL-5和IL-8的含量顯著下降,差異有統(tǒng)計學(xué)意義(P0.05)；肺組織GRmRNA的表達(dá)無明顯變化,HSP90和p38MAPK mRNA的表達(dá)顯著下降,差異有統(tǒng)計學(xué)意義(P0.05)；肺組織GR、HSP90和p38MAPK蛋白表達(dá)呈現(xiàn)與mRNA表達(dá)相一致的結(jié)果。C組A549細(xì)胞GR的核漿比為0.08±0.05,D組A549細(xì)胞GR的核漿比為0.59±0.25,兩組之間的差異有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論p38MAPK抑制劑SB203580可以改善煙霧暴露的哮喘大鼠的氣道炎癥,減輕其支氣管收縮反應(yīng)。其通過促使GR核轉(zhuǎn)位提高糖皮質(zhì)激素敏感性。
[Abstract]:Objective to establish a rat model of asthma induced by smoke exposure and an experimental model of nicotine induced glucocorticoid resistance in human lung adenocarcinoma cell line A549, and to observe the effect of p38 mitogen activated protein kinase (p38 MAPK) inhibitor SB203580 on smog exposed asthmatic rats.The effects of SB203580 on glucocorticoid sensitivity were investigated in vivo and in vitro.Methods Wistar rats were randomly divided into 4 groups: normal control group, asthma group, smoke exposed asthma group and SB203580 intervention group.The expiratory resistance, inspiratory resistance and lung compliance in rats were measured by animal pulmonary function analyzer. The histopathological changes of lung tissue were observed. The levels of IL-4 IL-5 and IL-8 in rat lung homogenate were detected by ELISA.The expression of glucocorticoid receptor (GRN) HSP90 and p38MAPK mRNA in rat lung tissue was detected by RT-PCR, and the expression of GRN HSP90 and p38MAPK protein in rat lung tissue was detected by Western blot.A549 cells were cultured,灝嗗叾闅忔満鍒嗕負(fù)鍥涚粍:A緇，

本文編號：1713147