Investigation the role of nitric oxide pathway in TNF-& alpha; induced HUVEC cells

Nitric oxide (NO) is a highly reactive molecule involved in a variety of physiological and pathophysiological processes, such as inflammation. eNOS-produced NO has anti-inflammatory properties and play a role in vascular homeostasis. Through a variety of mechanisms, tumor necrosis factor alpha (TNF-a) has been shown to induce inflammation in HUVECs. The purpose of this study was to investigate the role of NO in HUVEC cells under inflammatory conditions induced by TNF-a. To identify TNF-a induction mechanisms, the phosphorylation of ERK, Akt, and eNOS was investigated. Sildenafil and L-NAME used to examine the role of NO in this process. In TNF-ainduced HUVEC cells, cell viability, nitrite/nitrate production, phosphorylated and total ERK, Akt, and eNOS levels were measured with or without sildenafil and L-NAME. At 20 and 40 ng/ml concentrations, TNF-a increased nitrite/nitrate and decreased cell viability (p<0.05). Sildenafil and L-NAME had no effect on cell viability and they both decreased nitrite/ nitrate in HUVEC that had been stimulated by TNF-a. TNF-a had no effect on the phosphorylation of ERK, Akt, and eNOS, but it did increase total eNOS levels. The phosphorylation of ERK, Akt, and eNOS was unaffected by sildenafil and L-NAME; however, L-NAME decreased total eNOS. According to our findings, there is no known direct relationship between TNF-a, sildenafil, or L-NAME and protein phosphorylation.

Automated summary

The study investigated the role of nitric oxide (NO) in HUVEC cells under inflammatory conditions induced by tumor necrosis factor alpha (TNF-a). The phosphorylation of ERK, Akt, and eNOS was examined to identify TNF-a induction mechanisms. Sildenafil and L-NAME were used to examine the role of NO. The results showed that TNF-a increased nitrite/nitrate production and decreased cell viability, while sildenafil and L-NAME had no effect on cell viability but decreased nitrite/nitrate levels in stimulated HUVEC cells. TNF-a had no effect on the phosphorylation of ERK, Akt, and eNOS, but increased total eNOS levels. Sildenafil and L-NAME had no effect on protein phosphorylation.