Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: RESULT (part 4)

Primers specific for beta-actin (positive control, lane 2) and negative control (lane 3) were used for comparison. Before LPS infusion, 15 mins or even 25 mins after LPS infusion (data not shown), NOS-II-specific product detected by RT-PCR was absent in the lung  (Figure 5). NOS-II-specific 384 base pair RT-PCR product was seen only after 30 mins (Figure 4,5). To identify the lung cell type expressing NOS-III mRNA, in situ hybridization histochemistry with specific 693 base pair DIG-labelled cDNA probe was performed on sections of rat lung. Buy Asthma Inhalers Online Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury

Figure 5 Time course of lipopolysaccharide- (LPS) induced changes in mRNA expression of inducible nitric oxide synthase (NOS-II) in rat lung tissue. Rats that were untreated or treated with LPS were decapitated, and their lungs were harvested at the times indicated. Total RNA was prepared and assayed by reverse transcription polymerase chain reaction (RT-PCR) using gene-specific primers. A single 384 base pair (bp) product was amplified with NOS-II gene-specific primers (lanes 6 and 14) 30 and 120 mins after LPS infusion; no PCR product was observed in lane 3 or 10, ie, 0 and 15 mins after LPS administration. The expected 308 bp product of beta-actin was observed (positive control, lanes 1, 4, 8 and 11). No PCR product was obtained in the absence of any primers (negative control, lanes 2, 5, 9 and 13). Marker M1 is shown in lane 7 (pUC19/MspI with bands 501/489, 404, 331, 242 and 190 bp long). The size of the predicted amplified products is indicated on the left and right

This entry was posted in Lung injury and tagged 1-Amino-2-hydroxy-guanidine, Endotoxic shock, NG-nitro-L-arginine, Nitric oxide, Nitric oxide synthase type II, Nitric oxide synthase type III, S-nitroso-N-acetyl-penicillamine.