肺纤维化形成过程中肺泡Ⅱ型上皮细胞对肺成纤维细胞的影响及相关机制的探讨

The effect of alveolar typeⅡcell to lung fibroblast in the rat model of pingyangmycin induced pulmonary fibrosis

目的:探讨肺间质纤维化形成过程中肺泡Ⅱ型上皮细胞对肺成纤维细胞FSP1,TGF-β,HGFmRNA表达的影响和调控作用机制。方法:28只SD大鼠随机分为平阳霉素组和生理盐水组,平阳霉素组气管内给药复制大鼠肺纤维化动物模型,生理盐水组气管内注射等量生理盐水。分别于7,28天各处死平阳霉素组大鼠10只,生理盐水组大鼠4只,其中平阳霉素组3只及生理盐水组3只提取肺泡Ⅱ型上皮细胞,平阳霉素组6只提取肺成纤维细胞,分别进行培养。至2种细胞到亚汇合状态时,实验组即:用平阳霉素组肺泡Ⅱ型上皮细胞上清液培养平阳霉素组肺成纤维细胞48h;对照组即:用生理盐水组肺泡Ⅱ型上皮细胞上清液培养平阳霉素组肺成纤维细胞48h,然后用半定量RT-PCR法检测肺成纤维细胞FSP1,TGF-β,HGFmRNA表达量的变化。平阳霉素组及生理盐水组各余1只用作HE染色。结果:(1)实验组7天,28天FSP1mRNA,TGF-βmRNA,HGFmRNA表达均高于对照组(P<0.01),(2)第7,28天实验组FSP1mRNA,FGF-βmRNA表达无明显差别(P>0.05),实验组28天HGFmRNA表达高于第7天(P<0.01)。结论:平阳霉素诱导的肺纤维化大鼠的肺泡Ⅱ型上皮细胞能够促进怖成纤维细胞FSP1mRNA、TGF-βmRNA的表达,能够促进纤维化的进展。

Objective：To investigate the FSP1mRNA,TGF-β mRNA and HGFmRNA expression of lung fibroblasts being affected by alveolar type Ⅱ cells in pingyangmycin-induced pulmonary fibrosis in rats and its possible mechanism. Methods：Twenty-eight SD rats were randomly divided into two groups as the Pingyangmycin group and saline group . Pulmonary fibrosis was induced by intratracheal instillation with pingyangmycin in the Pingyangmycin group , and the saline group was treated with normal saline. Then ten rats in Pingyangmycin group and four rats in saline group were sacrificed respectively at the 7th,28th day after intratracheal instillation. Three sacrificed rats in the Pingyangmycin group and three sacrificed rats in the saline group were abstracted alveolar type Ⅱ cells. The six sacrificed rats in the Pingyangmycin group were abstracted lung fibroblasts. When the two kinds of cells were at the good state,the experiment group：put the supernate fluid of alveolar type Ⅱ cells of Pingyangmycin group into the lung fibroblasts of Pingyangmycin group;the control group： put the supernate fluid of alveolar type Ⅱ cells of saline group into the lung fibroblasts of Pingyangmycin group, then cocuhure 48 hours,the levels of FSP1mRNA,TGF-βmRNA,HGFmRNA of lung fibroblasts were detected by demi-quantitate RT-PCR. The rest rats of Pingyangmycin group and the saline group were used HE stained. Results： （1）The expression of FSP1mRNA, TGF-β mRNA, HGFmRNA in experiment group were higher than the control group at the 7 th,28th day （P〈0.01）. （2）The levels of FSP1mRNA,TGF-βmRNA in experiment group were not significantly difference between at the 7 th,28th day （P〉 0.05）,except the level of HGFmRNA in experiment group at the 28th was higher than at the 7th （P〈0.01）. Conclusion： Alveolar type Ⅱ cells in pingyangmycin-induced pulmonary fibrosis in rats can promote the FSP1mRNA,TGF-βmRNA expression of lung fibroblasts,and can enhance the procession of pulmonary fibrosis.