吸烟患COPD者与吸烟不患COPD者、肺组织中HIF-1α和MMP9表达的研究

发布时间：2018-04-10 09:11

本文选题：低氧诱导因子-1α　切入点：基质金属蛋白酶-9　出处：《华中科技大学》2014年博士论文

【摘要】：第一部分吸烟患COPD者与吸烟不患COPD者肺组织中HIF-1α和MMP9表达的研究目的：探讨吸烟患COPD者与吸烟不患COPD者及不吸烟者的肺组织中HIF-la和MMP9的表达以及相关性。方法：收集不吸烟不患COPD(对照组)、吸烟不患COPD(吸烟组)和吸烟且患COPD (COPD组)的患者的肺组织各20例,性别、年龄相匹配,用免疫组织化学法检测三组肺组织中HIF-1α和MMP9的表达部位,Western blotting法检测三组肺组织中HIF-1α和MMP9的蛋白表达量,明胶酶谱法测定三组肺组织中MMP9的活性。结果：相比于吸烟组和对照组,COPD组患者肺组织中HIF-1α和MMP9的细胞免疫组化染色阳性率最高,其中炎性细胞(主要为巨噬细胞等)、支气管上皮细胞等部位HIF-1α和MMP-9的表达均为阳性。COPD组肺组织中HIF-1α和MMP9的蛋白表达明显高于吸烟组,而吸烟组肺组织中HIF-1α和MMP9蛋白表达明显高于对照组。HIF-1α和MMP9的蛋白表达呈明显正相关。而且,COPD组肺组织中MMP9的活性明显高于吸烟组,而吸烟组肺组织中MMP9的活性明显高于对照组。结论：COPD组的肺组织中HIF-1α和MMP9的蛋白表达较不患COPD者明显增加,且两者的表达呈明显正相关。提示HIF-1α及MMP9可能参与吸烟致COPD发病机制的形成,两者之间可能有一定的关联性。第二部分在CSE和HIF-1α转录抑制剂作用下,HIF-1α和MMP9在肺泡巨噬细胞中表达的研究目的：探讨小鼠肺泡巨噬细胞在香烟烟雾提取物(CSE)刺激下,表达HIF-1α和MMP9的情况,以及HIF-1α的抑制对MMP9表达的影响。方法：灌注小鼠肺泡收集小鼠肺泡巨噬细胞,将其分为三组：对照组,CSE组,CSE+抑制剂组。用CSE单独或联合HIF-1α抑制剂CAY10585作用于小鼠肺泡巨噬细胞,Real-time PCR法检测三组肺泡巨噬细胞中HIF-1α和MMP9的mRNA表达量,Western blotting法检测三组肺泡巨噬细胞中HIF-1α和MMP9的蛋白表达量,明胶酶谱法测定三组肺泡巨噬细胞中MMP9的活性。结果：CSE组的肺泡巨噬细胞中MMP9的mRNA和蛋白表达量明显高于CSE+抑制剂组和对照组。CSE组中HIF-1α的mRNA表达量和CSE+抑制剂组没有明显差异,明显高于对照组。而CSE组中HIF-1α的蛋白表达量明显高于CSE+抑制剂组和对照组。HIF-1α和MMP9的蛋白表达呈明显正相关。而且,CSE组肺组织中MMP9的活性也明显高于其余两组。结论：肺泡巨噬细胞中,抑制HIF-1α的表达可以抑制MMP9的表达,且两者的表达呈明显正相关。提示HIF-1α的表达可能可以促进MMP9的表达。
[Abstract]:Part one study on the expression of HIF-1 伪 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 伪 and MMP9 in three groups of lung tissues was detected by immunohistochemical method. The expression of HIF-1 伪 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 伪 and MMP9 in lung tissue was the highest.The expression of HIF-1 伪 and MMP-9 in inflammatory cells (mainly macrophages and bronchoepithelial cells) was significantly higher than that in smoking group.The expression of HIF-1 伪 and MMP9 protein in lung tissue of smoking group was significantly higher than that of control group. HIF-1 伪 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 伪 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 伪 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 伪 and MMP9 in alveolar macrophages under the action of CSE and HIF-1 伪 transcription inhibitors.Aim: to investigate the expression of HIF-1 伪 and MMP9 in mouse alveolar macrophages stimulated by cigarette smoke extract (CSE) and the effect of inhibition of HIF-1 伪 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 伪 and MMP9 in alveolar macrophages of three groups were detected by CSE alone or in combination with HIF-1 伪 inhibitor CAY10585. The protein expression of HIF-1 伪 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 伪 between the control group and the group of CSE inhibitor, and the expression of HIF-1 伪 was significantly higher than that of the control group.The protein expression of HIF-1 伪 in CSE group was significantly higher than that in CSE inhibitor group and control group. HIF-1 伪 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 伪 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 伪 may promote the expression of MMP9.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R563.9

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