Expression Of K_ATP Channel In Rat And Research Of Efedrina On Smooth Muscle Cell Growth

Two methods of obtaining Artery Smooth Muscle Cell (ASMC) and Bronchus Smooth Muscle Cell (BSMC) were discussed and compared: explant culture and digest method. 0.2% Collagenase I was used to digest ASM for about 1hr in 37℃, followed by 0.05% Trypsin for 5-10min in RT. After 1 week, cells grew from the edge of original 1mm3 BSM explant, and they overspread after 3-4 weeks. Both ASMC and BSMC are zonary or spindly, with smooth membrane. They were subcultured in DMEM containing 10% FBS. To detect the expression of KATP channel subunits on rat airway smooth muscle cells and rat bronchia smooth muscle cells, RT-PCR was used. Total RNA is extracted from the 3rd-5th generation cells cultured, reversely translated, followed with PCR. Kir6.1 and SUR2B were found to be present on airway smooth muscle cells with SUR1 expressed in low quantity, and Kir6.1 and SUR2B were expressed on bronchia smooth muscle cells. Expected cDNA of Kir6.1 and SUR2B from both ASMC and BSMC have also been achieved. Ephedra is important in Respiratory Disease and Cardiovascular Disease. Ephedrine Hydrochloride and Pseudoephedrine Hydrochloride inhibited ASMC growth by 39.6% and 42.7% after 48hrs. Ephedrine Hydrochloride inhibited BSMC growth by 7.2% after 72 hrs, while Pseudoephedrine Hydrochloride had no effect.