Study On Aitc Upregulating MRP1 And Nrf2 Expression In Pulmonary Bronchial Epithelial Of Copd

OBJECTIVE To analyze the effect of AITC on MRP1 and Nrf2 expression in pulmonary bronchial epithelial of COPD,we use in vivo and in vitro COPD model.METHODS In vitro: First, we select a appropriate CSE concentration via MTT assay; After stimulating 16 HBE cell with CSE, real time RT-PCR was used to analyze MRP1 and Nrf2 m RNA expression. Western blotting was used to analyze MRP1 and Nrf2 protein levels. Then, we retreatment 16 HBE with AITC before stimulating with CSE, real time RT-PCR was used to analyze MRP1 and Nrf2 m RNA expression. Western blotting was used to analyze MRP1 and Nrf2 protein levels. In vivo: We establish a COPD model through cigarette smoking and intratracheal instillation of LPS in rats. After treatment with AITC, We inspect the general condition of rats and examine lung function and pulmonary pathological changes of rats. Then, real time RT-PCR was used to analyze MRP1 and Nrf2 m RNA expression in rats lung tissues.RESULTS1. The method to prepare CSE is quite standard and normative.2. Along with the increase of CSE concentration and the time of incubation, the inhibitory rates of CSE to 16 HBE cell increased via the MTT assay. When the CSE concentration was lower than 10%, the inhibitory rates were significantly lower than those when the concentrations were higher than 25%. There were no obvious difference between 12 h and 24 h incubation.3. 5% CSE could decrease the MRP1 and Nrf2 m RNA and protein expression.4. Pretreatment with different concentration of AITC before stimulating 16 HBE cell with CSE could increase the MRP1 and Nrf2 m RNA and protein expression. 40μΜ AITC had an obvious difference(P<0.05).5. Using cigarette smoking and intratracheal instillation of LPS to establish rat COPD models. We analyze the lung function of them. There were significantly difference between COPD rats and normal rats(P<0.05). While after treatment with positive drug(NAC) or high/low dose of AITC, the indicators of lung function including FVC,FEV0.3%/ FVC, MMF, PEF, Cldyn were higher than those of COPD rats.We also inspect the pulmonary pathological changes of COPD rats. Compared with normal rats,there were inflammatory cells infiltrated, goblet cells increased in COPD rats. After treatment with NAC or AITC, all pathological changes in COPD rats were significantly reduced. 6.Compared with normal rats, the MRP1 and Nrf2 m RNA expression in lung tissue of COPD rats declined. After treatment with NAC or AITC, the MRP1 and Nrf2 m RNA expression in lung tissue of rats increased compared with COPD rats.CONCLUSIONS In pulmonary bronchial epithelial of COPD, the MRP1 and Nrf2 expression both decreased. While AITC can up-regulate MRP1 and Nrf2 expression. AITC up-regulate MRP1 expression may be associated with Nrf2.