本文選題：低氧誘導因子-1α　切入點：基質(zhì)金屬蛋白酶-9　出處：《華中科技大學》2014年博士論文

【摘要】：第一部分吸煙患COPD者與吸煙不患COPD者肺組織中HIF-1α和MMP9表達的研究 目的：探討吸煙患COPD者與吸煙不患COPD者及不吸煙者的肺組織中HIF-la和MMP9的表達以及相關(guān)性。 方法：收集不吸煙不患COPD(對照組)、吸煙不患COPD(吸煙組)和吸煙且患COPD (COPD組)的患者的肺組織各20例,性別、年齡相匹配,用免疫組織化學法檢測三組肺組織中HIF-1α和MMP9的表達部位,Western blotting法檢測三組肺組織中HIF-1α和MMP9的蛋白表達量,明膠酶譜法測定三組肺組織中MMP9的活性。 結(jié)果：相比于吸煙組和對照組,COPD組患者肺組織中HIF-1α和MMP9的細胞免疫組化染色陽性率最高,其中炎性細胞(主要為巨噬細胞等)、支氣管上皮細胞等部位HIF-1α和MMP-9的表達均為陽性。COPD組肺組織中HIF-1α和MMP9的蛋白表達明顯高于吸煙組,而吸煙組肺組織中HIF-1α和MMP9蛋白表達明顯高于對照組。HIF-1α和MMP9的蛋白表達呈明顯正相關(guān)。而且,COPD組肺組織中MMP9的活性明顯高于吸煙組,而吸煙組肺組織中MMP9的活性明顯高于對照組。 結(jié)論：COPD組的肺組織中HIF-1α和MMP9的蛋白表達較不患COPD者明顯增加,且兩者的表達呈明顯正相關(guān)。提示HIF-1α及MMP9可能參與吸煙致COPD發(fā)病機制的形成,兩者之間可能有一定的關(guān)聯(lián)性。 第二部分在CSE和HIF-1α轉(zhuǎn)錄抑制劑作用下,HIF-1α和MMP9在肺泡巨噬細胞中表達的研究 目的：探討小鼠肺泡巨噬細胞在香煙煙霧提取物(CSE)刺激下,表達HIF-1α和MMP9的情況,以及HIF-1α的抑制對MMP9表達的影響。 方法：灌注小鼠肺泡收集小鼠肺泡巨噬細胞,將其分為三組：對照組,CSE組,CSE+抑制劑組。用CSE單獨或聯(lián)合HIF-1α抑制劑CAY10585作用于小鼠肺泡巨噬細胞,Real-time PCR法檢測三組肺泡巨噬細胞中HIF-1α和MMP9的mRNA表達量,Western blotting法檢測三組肺泡巨噬細胞中HIF-1α和MMP9的蛋白表達量,明膠酶譜法測定三組肺泡巨噬細胞中MMP9的活性。 結(jié)果：CSE組的肺泡巨噬細胞中MMP9的mRNA和蛋白表達量明顯高于CSE+抑制劑組和對照組。CSE組中HIF-1α的mRNA表達量和CSE+抑制劑組沒有明顯差異,明顯高于對照組。而CSE組中HIF-1α的蛋白表達量明顯高于CSE+抑制劑組和對照組。HIF-1α和MMP9的蛋白表達呈明顯正相關(guān)。而且,CSE組肺組織中MMP9的活性也明顯高于其余兩組。 結(jié)論：肺泡巨噬細胞中,抑制HIF-1α的表達可以抑制MMP9的表達,且兩者的表達呈明顯正相關(guān)。提示HIF-1α的表達可能可以促進MMP9的表達。
[Abstract]:Part one study on the expression of HIF-1 偽 and MMP9 in lung tissues of smoking patients with COPD and smoking without COPDObjective: to investigate the expression and correlation of HIF-la and MMP9 in lung tissues of smoking patients with COPD and non smoking patients with COPD and non smokers.Methods: lung tissues were collected from 20 non-smoking and non-smoking patients (control group, smoking group, smoking group and COPD group), and matched by sex and age.The expression of HIF-1 偽 and MMP9 in three groups of lung tissues was detected by immunohistochemical method. The expression of HIF-1 偽 and MMP9 in three groups of lung tissues was detected by Western blotting, and the activity of MMP9 in three groups of lung tissues was detected by gelatinase spectrum.Results: compared with smoking group and control group, the positive rate of HIF-1 偽 and MMP9 in lung tissue was the highest.The expression of HIF-1 偽 and MMP-9 in inflammatory cells (mainly macrophages and bronchoepithelial cells) was significantly higher than that in smoking group.The expression of HIF-1 偽 and MMP9 protein in lung tissue of smoking group was significantly higher than that of control group. HIF-1 偽 and MMP9 protein expression were positively correlated.The activity of MMP9 in lung tissue of smoking group was significantly higher than that of smoking group, while the activity of MMP9 in lung tissue of smoking group was significantly higher than that of control group.Conclusion the protein expression of HIF-1 偽 and MMP9 in the lung tissue of the group with COPD was significantly higher than that in the group without COPD, and there was a significant positive correlation between the expression of the two proteins.It is suggested that HIF-1 偽 and MMP9 may be involved in the pathogenesis of COPD induced by smoking, and there may be a certain correlation between them.The second part is about the expression of HIF-1 偽 and MMP9 in alveolar macrophages under the action of CSE and HIF-1 偽 transcription inhibitors.Aim: to investigate the expression of HIF-1 偽 and MMP9 in mouse alveolar macrophages stimulated by cigarette smoke extract (CSE) and the effect of inhibition of HIF-1 偽 on the expression of MMP9.Methods: alveolar macrophages were collected and divided into three groups: control group (CSE group) and CSE inhibitor group.The mRNA expression of HIF-1 偽 and MMP9 in alveolar macrophages of three groups were detected by CSE alone or in combination with HIF-1 偽 inhibitor CAY10585. The protein expression of HIF-1 偽 and MMP9 in alveolar macrophages of three groups were detected by Western blotting.The activity of MMP9 in three groups of alveolar macrophages was determined by gelatinase method.Results the expression of mRNA and protein of MMP9 in alveolar macrophages in the group of 1: CSE was significantly higher than that in the group of CSE inhibitor and the control group. There was no significant difference in mRNA expression of HIF-1 偽 between the control group and the group of CSE inhibitor, and the expression of HIF-1 偽 was significantly higher than that of the control group.The protein expression of HIF-1 偽 in CSE group was significantly higher than that in CSE inhibitor group and control group. HIF-1 偽 and MMP9 protein expression were positively correlated.The activity of MMP9 in lung tissue was significantly higher in CSE group than in other two groups.Conclusion: inhibiting the expression of HIF-1 偽 in alveolar macrophages can inhibit the expression of MMP9, and there is a positive correlation between them.These results suggest that the expression of HIF-1 偽 may promote the expression of MMP9.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R563.9

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本文編號：1730584