Abhd2基因与肺气肿发病机制研究

Abhd2 Genes and Emphysema Pathogenesis Research

目的:利用基因敲除技术构建的小鼠肺气肿模型研究Abhd2基因与肺气肿发病机制的相关性。方法:生后6月龄,12月龄的Abhd2基因敲除纯合子和野生型鼠各5只。断髓法处死小鼠,取全肺,提取肺总RNA,紫外线分光光度计测定肺总RNA纯度并计算其浓度。PCR扩增产物行1.5%的琼脂糖凝胶电泳,在紫外线凝胶扫描仪观察拍照,存入IDKadak成像分析系统,分别读取肺气肿的相关因子和相应β-actin光密度值。电泳带密度运用ImageJ软件通过光密度测定法定量分析,比值用于统计学分析。结果:12个月龄Abhd2基因敲除纯合子小鼠与野生型对照比肺组织中基质金属蛋白酶9、12、13、14、组织蛋白酶B、K、S的mRNA表达水平明显增高,金属蛋白酶组织抑制剂3mRNA表达水平明显下降,氧化剂与抗氧化剂的mRNA表达水平未见异常,Abhd2基因敲除纯合子小鼠与野生型对照小鼠比肺组织中白细胞介素-lβ、白细胞介素-6、白细胞介素-13和肿瘤坏死因子-α的mRNA表达明显增高。结论:Abhd2基因敲除小鼠通过肺组织中巨噬细胞浸润增多、致炎细胞因子表达过多、蛋白酶基因表达过强与抗蛋白酶抑制剂表达降低以及异常增多的细胞凋亡,自发形成了肺气肿,且渐进性进展,而且肺气肿发生、发展过程与人类是相似的;因此它们对于人类肺气肿遗传易感性及环境因子的研究具有重要价值。

Objective： To investigate the relationship of Abhd2 genes and emphysema pathogenesis by using gene knock out mice model construction technology. Methods： 6 months and 12 months after birth the Abhd2 knockout homozygous form and wild type of 5 rat alone. Broken pulp method put to death, take the mice lung, total RNA, ultraviolet radiation extraction pulmonary lung spectrophotometer to measure and calculate the total RNA purity concentration. Amplified PCR product line 1.5% agarose gel electrophoresis, uv gel scanner observation in pictures and deposit IDKadak imaging analysis system, readed the related factors respectively emphysema and corresponding beta actin light density values. Electrophoresis using ImageJ software with density through the optical density measurement of quantitative analysis for statistical analysis, ratio. Results： 12 months Abhd2 gene knock out mice with homozygous wild type than controls lung of matrix metalloproteinases 9, 12, 13, 14, organization protease B, K, S mRNA expression level higher, metal protease inhibitors 3 mRNA expression level organization declined obviously, antioxidant and antioxidant mRNA expression level did not see, Abhd2 abnormal gene knock out mice with homozygous wild type than control mice lung of interleukin -16 , intefleukin-6, interleukin - 13 and pharmacological mRNA express significantly higher. Conclusion： Abhd2 knockout mice through the lung of macrophage inflammatory cells infiltrating increase, the factor expression too much, protease gene expressed a strong resistance and protease inhibitors decreased and the increase of abnormal express cell apoptosis, spontaneously formed emphysema, and incremental progress, and emphysema occurrence, development process and humans are similar; so they for human genetic predisposition and environmental factors of emphysema research is of important value.