Tag Archives: Nitric oxide synthase type II

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

RT-PCR technique was used to reveal the kinetics of NOS-II mRNA appearance in the lung. The 384 base pair single band was amplified from samples with NOS-II primers only 30 and 120 mins after LPS infusion, but not in controls or in rat pulmonary tissue treated for 15 mins and 25 mins (data not shown) with LPS . These data, together with results from dot … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: DISCUSSION (part 3)

Kinetics of appearance of NOS-II mRNA in several rat organs (lung, liver, heart, spleen and kidney) by dot blot analysis indicated that NOS-II transcripts appear in lung and kidney as early as 30 mins after LPS infusion. Two hours after LPS infusion, NOS-II mRNA was detected in all tested organs. We looked for confirmation of the unexpected finding by dot blot technique that pulmonary NOS-II … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: DISCUSSION (part 2)

When the preferential NOS-II inhibitor AGD (15 mg/kg intravenously) (Figure 2) was given instead of L-NNA before LPS infusion, survival time was lengthened compared with that of LPS-treated rats. These data suggest that the release of nitric oxide in the early phase of endotoxemia may prevent acute pneumotoxicity of LPS caused by platelet-activating factor, thromboxane A2 or sulphidopeptide leukotrienes . On the other hand, the … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: DISCUSSION (part 1)

Knowing the dosage of LPS and the mode of its administration (intraperitonial, intravenous, injection, infusion) is essential when studying the role of nitric oxide in endotoxe-mia. Some authorities in the field consider an initial fall in blood pressure occurring some minutes after administration of LPS to be an epiphenomenon – its mechanism not worthy of study. This reasoning seems correct when directly transferring experimental results … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: RESULT (part 9)

Only a few positive cells were found in lung sections from rats treated for 30 mins with LPS (Figure 10), with localization apparently different from those reacting with NOS-III-specific probe. Microscopic examination under a higher magnification (Figure 11) revealed that pulmonary vessels were filled with positive-stained cells, which may suggest that the cellular source of NOS-II, even at the beginning of the second phase of … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: RESULT (part 8)

As expected, NOS-II mRNA was detected in LPS-treated rats but not in control rats (Figure 9).  These data together with the results of RT-PCR  and dot blot analyses  suggest that NOS-II transcripts appeared in endotoxemic rat lung not earlier than 30 mins after LPS infusion. This is why the nitric oxide that is generated by LPS from NOS-III but not NOS-II is one of the … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: RESULT (part 7)

NOS-II mRNA was detected in all tested organs of LPS-treated rats after 2 h. The NOS-II transcripts appeared as early as 30 mins after LPS infusion in the lung and kidney (Figure 8). To ensure that it was possible to detect NOS-II transcript in lung tissue treated for 30 mins with LPS, Northern blot analysis was performed using a DIG-labelled 473 base pair cDNA probe. … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: RESULT (part 6)

Identification of NOS-II transcripts and kinetics of their appearance: Freshly homogenized lung tissue from controls and from LPS-treated rats (decapitated 15, 30 and 120 mins after LPS infusion) were subjected to total RNA isolation by the Trizol method. RNA (1 |ig) was reverse transcribed and amplified by PCR with rat-specific oligonucleotide primers for NOS-II (Figure 5). Beta-actin-specific primers were used as a positive control. Negative control … Continue reading

Role of nitric oxide synthase types II and III in early protection against endotoxin-induced lung injury: RESULT (part 5)

Similarly, by using a specific polyclonal antibody against NOS-III, the immunohistochemical localization of NOS-III antigen followed the signal that was obtained by the in situ hybridization technique (Figure 7). NOS-III mRNA was intensely and specifically visualized in endothe-lial cells of both control (data not shown) and LPS-treated rat lung (Figure 6). ventolin inhaler

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 … Continue reading