HO-1、NF-κB在矽肺纤维化中作用机制的实验研究

The role of HO-1 and NF-κB in the formation of experimental silicosis of rats

目的观察血红素加氧酶-1（HO-1）及核转录因子kappa B（NF-κB）在大鼠矽肺纤维化中的作用机制及相互关系。方法实验动物采用Wistar大鼠60只,体重180~220g,雌雄不拘,适应性饲养1周后,随机分为4组,分别为对照组（A组）和染尘模型组（B组）、染尘＋HO-1抑制剂组（C组）、染尘＋HO-1诱导剂组（D组）,每组15只。采用气管暴露法滴注二氧化硅悬液制造大鼠矽肺模型。建模28d后HE染色观察各组大鼠肺组织病理结构,采用Western blot法检测各组肺组织HO-1和NF-κB蛋白含量,采用荧光定量PCR法检测各组肺组织HO-1和NF-κB mRNA的表达。结果 Western blot检测结果显示HO-1、NF-κB蛋白含量,对照组明显低于其他各组（P〈0.05）,C组较B组明显降低,而D组较B组表达量明显升高（P〈0.05）。RT-PCR检测各组HO-1mRNA相对表达量分别为2.25±0.38、1.81±0.74、7.75±0.56,C组较B组表达明显降低,而D组较B组表达显著升高（P〈0.05）;NF-κB mRNA相对表达分别为3.06±0.42、6.15±0.37、1.39±0.14,C组较B组表达明显升高,而D组较B组表达显著降低（P〈0.05）。结论 HO-1、NF-κB共同参与了大鼠矽肺纤维化的形成过程,通过对其干预可以改变大鼠矽肺纤维化的程度,可为矽肺纤维化的诊断与治疗提供新途径。

Objective To observe the regulatory function of heme oxygenase 1（HO-1）and nuclear factor Kappa B（NF-κB）on the formation of pneumonoconiosis in rats. Methods Sixty Wistar rats weighted 80-220 g were randomly divided into 4groups after a week adaptive breeding.Rats in group A served as controls,while those in other groups were treated with dust（group B）,plus HO-1inhibitor zinc protoporphyrin（group C）,or HO-1inducer cobalt protoporphyrin（group D）.Twenty-eight days after the rat model of silicosis was established,all rats were sacrificed and pathological changes of lung tissues were observed.Meanwhile,the expressions of HO-1and NF-κB in lung tissues of rats were detected using Western blot and real time quantitative PCR. Results The expressions of HO-1and NF-κB in lung tissues were the highest in group D,followed by those in group B,and the lowest in control group（P〈0.05）.RT-PCR revealed that HO-1mRNA increased significantly from 2.25±0.38 in group B to 7.75±0.56 in group D,while it was the lowest of 1.81±0.74 in group C rats（P〈0.05）.However,NF-κB mRNA was found to be the highest in group C（6.15±0.37）and the lowest in group D（1.39±0.14）,it was 3.06±0.42 in rats of group B,the differences were statistically significant（P〈0.05）. Conclusions Both HO-1and NF-κB involve in the process of idiopathic pulmonary fibrosis in rats,indicating that they may serve as intervention targets to provide new ways for pulmonary fibrosis diagnosis and therapy.