Investigate The Mechanism Of Allyl Isothiocyanate In Treatment Of Copd Based On Its Pharmacokinetics And The Effect On The Function Of MRP1

Objective To study therapeutic effect for chronic obstructive pulmonary disease(COPD) rat model of Allyl isothiocyanate, the main component of the volatile oil fromSemen Sinapis Albae. The molecular mechanism of allyl isothiocyanate in COPD istreating is clarified according to study the pharmacokinetics of allyl isothiocyanate in ratand the impact of allyl isothiocyanate on function and expression of multidrugresistance-associated protein1(MRP1) in human bronchial epithelial cells.Methods1) The essential oil was extracted by steam distillation. The content of allylisothiocyanate was determined by GC.2) Make quantitative stimulation with tobacco，SO2and papin to establish COPD rat model, lung function measurements were carriedout before and after the treatment of allyl isothiocyanate.3) Ultra performance liquidchromatographic(UPLC) method was established for detecting allyl isothiocyanate ofbiological samples. After intravenous injection and oral administration of allylisothiocyanate, the absolute bioavailability and pharmacokinetic parameters of rats andthe main tissue distribution of rats were estimated.4) Human bronchial epithelial cellswere cultured. MTT assay was used to detect the inhibitory effect of allyl isothiocyanateon human bronchial epithelial cells. RT-PCR, Western blot and flow cytometry methodwere used to detect the effect of allyl isothiocyanate on MRP1in human bronchialepithelial cells.Results1) To establish the method for the content determination of allylIsothiocyanate in Semen Sinapis Albae by GC. Allyl Isothiocyanate has good linerrelationship in the concentration range of10～100μg/ml (r=0.9996). The averagerecovery was97.64%, RSD was2.39%. The content of7.36%.2）Compared withnormal group, the CL、FEV0.3/FVC、PEF and MMF of model rats were significantlylower (P <0.05). Compared with model group, the CL、FEV0.3/FVC、PEF and MMF of treatment group were significantly increased (P <0.05).3) An UPLC was established fordetecting for allyl isothiocyanate in biological samples. There was a linear relationshipof the concentration of allyl isothiocyanate within the range of0.156to40.0μg/mL (r=0.9989). The recovery was91.34%. The lower limit of quantitation of this method was0.156μg/mL. The elimination half life was77.36±8.43min, and the clearance was9.73±0.68mL/min, and the mean retention time was109.22±15.54min, and the area undercurve(AUC0→t) was2272.04±111.06min·μg/mL in rats with intravenous injection of20mg/kg allyl isothiocyanate. The elimination half life was107.46±5.42min, and theclearance was0.01±0.0075L/min/kg, and the mean retention time was323.14±2.31min,and the area under curve(AUC0→t) was1891.65±87.36min·μg/mL in rats with oraladministration of20mg/kg allyl isothiocyanate. The absolute bioavailability was84.95%. Allyl isothiocyanate was widely distributed in rat tissues after intravenousinjection administration of allyl isothiocyanate, mainly in the kidney, lung, liver,stomach, heart, spleen, muscle, brain.4）We evaluated the inhibitory effect of AITC onhuman bronchial epithelial by using the MTT assay. The IC50of AITC was35.86±0.037μM in16HBE14o-cells. It was found that AITC (1～40μM) caused a change ofmRNA levels in MRP1and AITC (5～40μM) increased the change of protein level andAITC treatment of16HBE14o-cells at concentration of5～40μM resulted insignificantly increased in MRP1function.Conclusions Allyl isothiocyanate has therapeutic effect in treating COPD byimproving the lung function of COPD rat model. Its therapeutic effect related to thatallyl isothiocyanate had a higher bioavailability in rats and widely distributed in thelung tissue, as well as up-regulated the function and expression of MRP1in humanbronchial epithelial cells.