【摘要】：目的：探討吸煙伴或不伴有慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)患者肺血管重塑中細(xì)胞外信號(hào)調(diào)節(jié)激酶(extracellular signal-regulated kinase, ERK)的表達(dá)及意義。 方法：取48份(吸煙患COPD組16例、吸煙不患COPD組18例、不吸煙不患COPD組14例)手術(shù)切除的肺組織,HE染色觀察肺血管重塑；用Western blot檢測(cè)ERK及cyclinD1在肺動(dòng)脈的表達(dá)；實(shí)時(shí)熒光RT-PCR檢測(cè)cyclinD1mRNA的表達(dá)。 結(jié)果：與非吸煙對(duì)照組比較,吸煙不伴COPD組肺血管壁厚度和血管直徑比值(%WT)明顯增加,ERK及cyclinD1表達(dá)增加,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。吸煙不伴COPD組與吸煙伴COPD組相比較,肺血管壁厚度和血管直徑比值及ERK的表達(dá)差異無統(tǒng)計(jì)學(xué)意義。 結(jié)論：吸煙伴或不伴有COPD患者肺動(dòng)脈ERK和cyclinD1的表達(dá)均較不吸煙者明顯升高,ERK的高表達(dá)可能與香煙煙霧導(dǎo)致的肺血管重構(gòu)有關(guān)。 目的：探討細(xì)胞外信號(hào)調(diào)節(jié)激酶(extracellular signal-regulated kinase, ERK)在香煙煙霧提取物(cigarette smoke extract, CSE)所致人肺動(dòng)脈平滑肌細(xì)胞(human pulmonary artery smooth muscle cells, HPASMCs)增殖中的作用。 方法：原代培養(yǎng)的HPASMCs予以CSE干預(yù)。細(xì)胞隨機(jī)分為四組：正常對(duì)照組；5%CSE組；PD98059+5%CSE組；PD98059組。采用細(xì)胞計(jì)數(shù)及四甲基偶氮唑鹽比色法(MTT)檢測(cè)細(xì)胞的增殖能力。流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期的分布。用Western blot檢測(cè)ERK及cyclinD1在HPASMCs的表達(dá)。實(shí)時(shí)熒光RT-PCR檢測(cè)cyclinDl mRNA的表達(dá)。 結(jié)果：在5%CSE的刺激下,HPASMCs的ERK mRNA和蛋白水平均升高,并促進(jìn)細(xì)胞增殖(P＜0.01)。PD98059抑制5%CSE誘導(dǎo)的細(xì)胞增殖(P＜0.01)。此外,PD98059明顯抑制cyclinDl的mRNA和蛋白的表達(dá),并導(dǎo)致細(xì)胞周期G1期停滯,進(jìn)而減少HPASMCs的增殖(P0.01)。 結(jié)論：ERK在CSE誘導(dǎo)HPASMCs增殖中發(fā)揮重要作用,其部分原因可能是其上調(diào)cyclinD1的表達(dá)。 目的：探討還原型輔酶Ⅱ(nicotinamide adenine dinucleotide phosphate, NADPH)氧化酶在香煙煙霧提取物(cigarette smoke extract, CSE)所致人肺動(dòng)脈平滑肌細(xì)胞(human pulmonary artery smooth muscle cells, HPASMCs)增殖中的作用。 方法：原代培養(yǎng)的HPASMCs予以CSE干預(yù)。細(xì)胞隨機(jī)分為四組：正常對(duì)照組；5%CSE組；apocynin+5%CSE組;apocynin組。采用細(xì)胞計(jì)數(shù)及四甲基偶氮唑鹽比色法(MTT)檢測(cè)細(xì)胞的增殖能力。流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期的分布。用Western blot檢測(cè)NADPH氧化酶亞基p47phox及細(xì)胞外信號(hào)調(diào)節(jié)激酶(extracellular signal-regulated kinase, ERK)在HPASMCs的表達(dá)。實(shí)時(shí)熒光RT-PCR檢測(cè)p47phox mRNA的表達(dá)。比色法檢測(cè)丙二醛(malondiadehyde, MDA)表達(dá)。 結(jié)果：在5%CSE的刺激下,HPASMCs中的p47phox以及ERK表達(dá)水平升高,并促進(jìn)細(xì)胞增殖(P＜0.01)。Apocynin抑制5%CSE誘導(dǎo)的細(xì)胞增殖(P＜0.01)。此外,apocynin明顯抑制ERK的表達(dá),并導(dǎo)致細(xì)胞周期G1期停滯,進(jìn)而減少HPASMCs的增殖(P＜0.01)。 結(jié)論：NADPH氧化酶作為ERK的上游信號(hào)通路可能在CSE誘導(dǎo)HPASMCs增殖中發(fā)揮了重要的調(diào)控機(jī)制。
[Abstract]:Aim: to investigate the expression and significance of extracellular signal regulated kinase (extracellular signal-regulated kinase, ERK) in pulmonary vascular remodeling in patients with or without chronic obstructive pulmonary disease (chronic obstructive pulmonary disease, COPD). Methods: 48 surgically resected lung tissues (16 cases with COPD, 18 cases with COPD without smoking and 14 cases with no smoking without COPD) were taken to observe the pulmonary vascular remodeling by HE staining, and the expression of ERK and cyclinD1 in pulmonary artery were detected by Western blot. The expression of cyclinD1mRNA was detected by real-time fluorescence RT-PCR. Results: compared with the non-smoking control group, the pulmonary vascular wall thickness and vascular diameter ratio (% WT) and the expression of ERK and cyclinD1 were significantly increased in the non-smoking group without COPD (P0.01). There was no significant difference in pulmonary vascular wall thickness, vessel diameter ratio and ERK expression between smoking without COPD group and smoking with COPD group. Conclusion: the expression of ERK and cyclinD1 in pulmonary artery of smoking patients with or without COPD is significantly higher than that of non-smokers. The high expression of ERK may be related to pulmonary vascular remodeling induced by cigarette smoke. Aim: to investigate the role of extracellular signal regulated kinase (extracellular signal-regulated kinase, ERK) in the proliferation of human pulmonary artery smooth muscle cells (human pulmonary artery smooth muscle cells, HPASMCs) induced by cigarette smoke extract (cigarette smoke extract, CSE). Methods: primary cultured HPASMCs were treated with CSE. Cells were randomly divided into four groups: normal control group; 5%CSE group; PD98059 5%CSE group; PD98059 group. Cell count and tetramethylazo salt colorimetric assay (MTT) were used to detect the proliferation ability of the cells. Cell cycle distribution was detected by flow cytometry. The expression of ERK and cyclinD1 in HPASMCs was detected by Western blot. The expression of cyclinDl mRNA was detected by real-time fluorescence RT-PCR. Results: under the stimulation of 5%CSE, the levels of ERK mRNA and protein of HPASMCs increased and promoted the proliferation of cells (P < 0 01). PD98059 inhibited 5%CSE induced cell proliferation (P < 0 01). In addition, PD98059 significantly inhibited the expression of mRNA and protein in cyclinDl, and led to cell cycle arrest in G1 phase, thus reducing the proliferation of HPASMCs (P0.01). Conclusion: ERK plays an important role in the proliferation of HPASMCs induced by CSE, which may be due to its up-regulation of cyclinD1 expression. Aim: to investigate the role of reduced coenzyme 鈪，

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