D4-GDI Is Up-regulated In Lung Tissue Of Passive Smoking Rat And RLE-6TN Cells Exposed To Cigarette Smoke Extract

[Objective] To explore the expression of D4-GDI in lung tissue of passivesmoking rat and RLE-6TN exposed to Cigarette Smoke extract, to verify whethersmoking cause expression of D4-GDI increased.[Methods]1.Animal model experiment: To establish the model of passive smoking rataccording to the traditional methods in the lab, measured pulmonary function. Tocollect the control groups, the2-mouth CS exposed groups,4-mouth CS exposedgroups, smoking cessation groups’lung tissue, expression of D4-GDI was detected byimmunohistochemistry, Reverse transcription-Polymerase Chain Reaction(RT-PCR)and Western blot.2.Cell experiment: RLE-6TN exposed to10%CSE for0h,3h,6h,12h,24h,expression of D4-GDI was detected by immunofluorescence technic, RT-PCR andWestern blot.[Results]1.Animal model experiment: RT-PCR results showed the expression of D4-GDIis the lowest in control groups(1.21±0.06), then followed by2-mouth CS exposedgroups(2.09±0.07), the highest in4-mouth CS exposed groups(2.45±0.12),smoking cessation groups(1.84±0.05)reduced but still higher than control groups,P<0.05, western blot results showed the expression level change trend of four groupswas accorded with the RT-PCR results, they respectively were(1.46±0.11、1.03±0.06、0.79±0.15、0.30±0.06), P<0.05, immunohistochemical staining showed thatD4-GDI expressed in variety of lung tissue cells except for bronchial epithelial cells,expression level change trend between groups was accorded with the Western blot results.2.Cell experiment: At different time point of RLE-6TN exposed to10%CSE,RT-PCR results showed that D4-GDI expression began to rise gradually from3hgroup(0.40±0.02), the12h group(0.69±0.05) was peak, then24h group(0.60±0.03)reduced but still higher than the6h group(0.52±0.04),0h group(0.31±0.01) was thelowest, P<0.05, western blot results showed that D4-GDI expression began to risegradually from3h group(0.78±0.08),6h group(1.11±0.12)、12h group were(1.65±0.13), the24h group(1.37±0.07) was peak,0h group(0.44±0.06) was the lowest,P<0.05, immunofluorescence staining results was accorded with the Western blotresults.[Conclusion]Smoking lead to expression of D4-GDI increased in rat lung tissue and AECII，D4-GDI may be involved in the pathogenesis of COPD.