| Objective: Study on the changes of multidrug resistance-associated protein1（MRP1）function in progression of chronic obstructive pulmonary disease (COPD) in pulmonarybronchial epithelial and the reverse effects of huatanjiangqi prescription on the functionof MRP1Methods:1) Establish the method of evaluation of the pulmonary bronchial epithelialMRP1function: The mice intravenous administered phenolsulfonphthalein (250mg/kg),pulmonary alveoli lavage after30min, the phenolsulfonphthalein concentration inBALF and plasma was mensurated, and inhalation of the MRP1specific inhibitorprobenecid (150mg/kg) as control, observed given the inhibitors before and after theconcentration of phenolsulfonphthalein in BALF/plasma ratio, to evaluate the changesof MRP1function in mice pulmonary epithelial cells.2) Study the relationship betweenprogression of COPD and the change of MRP1function in pulmonary bronchialepithelium: Make stimulation with tobacco and papain to establish COPD rat model.According to the principle of random, the rats were divided into the normal group,different course of COPD groups, probenecid control group and inhibitor groups. Theinhibitor groups were gave the MRP1inhibitor probenecid with the establishing COPDrat model. All the rats were evaluated the function of lung. All the rats were measuredthe permeability and integrity of the lung epithelial cells, expect the inhibitor group. Thephenolsulfonphthalein concentration in bronchoalveolar lavage fluid (BALF) andplasma was determined and the concentration of in BALF/plasma ratio was calculatedto evalutation of COPD lung bronchial epithelial MRP1function.3) Based on thechanges of MRP1function, we compared the characteristics of making stimulation withtobacco and papain and with lipopolysaccharide to establish COPD rat model.Usinglipopolysaccharide dropping method to establish COPD rat model, the rats wererandomly divided into process and model groups during the preparation. All the rats were taken the determination of pulmonary function; We measured the concentration ofphenolsulfonphthalein in BALF and the concentration of phenolsulfonphthalein inBALF/plasma ratio as the evaluation indicators of COPD pulmonary bronchialepithelial MRP1function. At the same time using immunohistochemistry to observe thechanges of pulmonary bronchial epithelial MRP1expression in tobacco and papainmodel group and lipopolysaccharide model group.4) The effect of huatanjiangqiprescription to the MRP1function changes of COPD model rats: Making stimulationwith tobacco and papain to establish COPD rat model. According to the principle ofrandom, we separated the rats into high dose group, middle dose group, low dose groupand positive control group. After drug treatment, all the rats were taken thedetermination of pulmonary function; We also measured the concentration ofphenolsulfonphthalein in BALF and plasma, the concentration of phenolsulfonphthaleinin BALF/plasma ratio as the evaluation indicators of COPD pulmonary bronchialepithelial MRP1function.5) Huatanjiangqi prescription had an effect on the excretionfunction of bronchial epithelial MRP1in vitro:16HBE14o-cells were cultured in vitro,5-CFDA as a substrate, determination of huatanjiangqi prescription and serumcontaining huatanjiangqi prescription on intracellular fluorescence intensity by flowcytometry instrument to evaluate the effect of drug and its metabolites on MRP1function.Results:1) Compared with the inhibitor is not given, to give the MRP1inhibitorprobenecid, plasma concentration of phenolsulfonphthalein was no significantdifference（P>0.05）. But not given inhibitor, after giving MRP1inhibitor probenecid60min, the concentration of phenolsulfonphthalein in BALF/plasma ratio was significantlydecreased（P<0.05）.2) With the development of disease, the lung functions of the ratsin different progression of COPD are gradually reduced. There is a significantdifference in45days later. With the process of COPD, the ratio ofphenolsulfonphthalein concentration in BALF and plasma are gradually reduced, but the lung epithelial cell permeability in each groups has not changed and the integrity of thelung epithelial cells is fine. Compared with normal groups, lung function of the inhibitorgroup (30days) is significantly reduced. There is a significant difference inFEV0.3%/FVC between the different courses of COPD groups and inhibitor groups in60days.3) We dropped lipopolysaccharide in rat bronchial to establish COPD modelrats.With the development of disease, the lung functions of the rats in differentprogression of COPD and model group were gradually reduced. the concentration ofphenolsulfonphthalein in BALF/plasma ratio were gradually reduced.Immunohistochemistry results showed that the MRP1protein in the model group oflipopolysaccharide and tobacco were weakly positive expression compared with thenormal group. There was no significant difference between two modeling methodsbased on the study of MRP1function in COPD condition or the drug affect the functionof MRP1.4) After treatment with huatanjiangqi prescription, the FEV0.3%/FVC、MMF、PEF and Cldyn of treatment group and positive control group rats weresignificantly higher（P<0.05) compared with the model group; After intravenousadministrated of phenolsulfonphthalein, the concentration of phenolsulfonphthalein inBALF/plasma ratio of treatment group and positive control group rats was graduallyincreased, which had a statistically significant(P<0.05) compared with the model group.5) Compared with the5-CFDA group, the intracellular fluorescence intensity ofhuatanjiangqi prescription and serum containing huatanjiangqi prescription becameweaker and had statistical significance(P<0.05).Conclusions: There is a certain correlation between the process of COPD and changesof MRP1function in lung bronchial epithelial. With the development of Chronicobstructive pulmonary disease, the efflux function of MRP1were gradually reduced.Inhibit the function of MRP1could aggravate the pulmonary function of COPD. At thesame time，this results illustrated that the MRP1played an important role in pulmonarybronchial epithelium cell. Huatanjiangqi prescription could improve the lung function in a rat COPD model. Its effect was correlated with regulating the function of MRP1inbronchial epithelium. |
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