| BackgroundInterstitial lung disease (interstitial lung disease, ILD) is a group of diffuse lung disease mainly involved alveoli, bronchioles and pulmonary interstitial tissue. The progress is slow, and gradual loss of lung function. Ultimately it develops into diffuse pulmonary fibrosis and honeycomb lung, leading to respiratory failure and death. Pulmonary fibrosis (pulmonary fibrosis, PF) is the most common interstitial lung disease, as well as the common end of a variety of lung diseases, because of its 5-year survival rate is low, so the concern of many scholars at home and abroad. At present the pathogenesis of pulmonary fibrosis mechanisms are not yet clear, most studies focus mainly on two aspects:one is the cell factor, the other is apoptosis. A variety of cells and cytokines participate in the pathogenesis of pulmonary fibrosis. Moderate apoptosis of these abnormal elements is conducive to the recovery of lung tissue structure, if insufficient apoptosis or excessive apoptosis may result in damage.of lung structure. Fas/FasL signal transduction pathway is the classical pathway of apoptosis, and it plays an important role in the pathogenesis of pulmonary fibrosis.Variety of experiments confirmed that in the fibrosis lung tissue, the expression of Fas/FasL upregulated in a variety of inflammatory cells,alveolar and bronchial epithelial cells, so it suggested that Fas/FasL-mediated apoptosis related to the occurrence of pulmonary fibrosis, In particular, the structure of lung tissue excessive apoptosis of cells, caused structural changes in lung tissue and thus led to pulmonary fibrosis. Soluble Fas antigen, anti-FasL antibodies could delay bleomycin-induced pulmonary fibrosis in rats, indicating that by inhibiting the Fas/FasL signaling pathway may have therapeutic value of pulmonary fibrosis.RNA interference (RNAinterference, RNAi) is a rapid development bio-engineering technology in recent years.It Refers to in the role of specific factors, by intracellular double-stranded into RNA (double strand RNA, dsRNA) degraded into small interfering RNA (small interfering RNA, siRNA), the siRNA with homologous complementary target RNA binding, specific enzyme Degradation of target RNA, thereby inhibiting, down target gene expression,.It is a new induced gene silencing.ObjectiveTo constructthe specific small interfereing RNA(siRNA) Duplication-deficiency adenovirus vector that can block the ratFasL gene according to the mRNA of rat FasL, To investigate the inhibitory effect of FasL mRNA transgenic expression on bleomycin-induced pulmonary fibrosis in mice.Materials and methods80 healthy SD rats were divided into control group, normal saline control group, FasL recombinant replication-deficient adenovirus treatment group, replication- defective adenovirus vector group, the three groups after were in 0d 0.4ml by intratracheal instillation of bleomycin (BLM 5 mg/kg) to prepare the interstitial lung fibrosis model,while the control group instilled the same volume of saline. Treatment group after instillation of bleomycin by nasal instillation of FasL 24h replication defective recombinant adenovirus (AdCMV FasL) 10×109 plaque forming units (pfu) (100μl)/mice, empty vector group in the trickle-down to Bo After the nasal instillation of amphotericin 24h replication-defective adenovirus vector (AdCMVNull), saline group at 24h after bleomycin instillation of nasal drops with the same volume of saline (100μl)/rat, control group 24h post-nasal drip with saline (100μl)/mice. The rats were first 7,14,28d were killed 5, the lung tissue hematoxylin eosin (HE) staining, was detected by immunohistochemistry in lung tissue levels of FasL protein expression, the evaluation by inhibiting the Fas/FasL pathway in pulmonary interstitial fibrosis.ResultThe alveolitis and fibrosis of the treatment group were less than that in normal saline group and sham recombinant adenoviruses group, in HE sections, the Less collagen deposition showed in treatment group compared to the normal saline group and sham recombinant adenoviruses group, and no collagen deposition showed in control group in Masson section. The immunohistochemistry expression of Fasl in treatment group was significantly lower than the normal saline group and sham recombinant adenoviruses group (P<0.05)ConclusionThe RNA interference mediated by Adenovirus vector can inhibit the expression of Fasl in lungs of mice with pulmonary fibrosis; and reduce pulmonary fibrosis, It is expected to be a way in the prevent and treat of pulmonary fibrosis. |
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