Folic acid supplementation repressed hypoxia-induced inflammatory response via ROS and JAK2/STAT3 pathway in human promyelomonocytic cells.

Hypoxia is associated with inflammation and various chronic diseases. Folic acid is known to ameliorate inflammatory reactions, but the metabolism of folic acid protecting against hypoxia-induced injury is still unclear. In our study, we examined the inflammatory signal transduction pathway in human promyelomonocytic cells (THP-1 cells) with or without treatment with folic acid under hypoxic culture conditions. Our results indicated that supplementation with folic acid significantly reduced the levels of interleukin-1ß and tumor necrosis factor-α in hypoxic conditions. Treating THP-1 cells with folic acid suppressed oxidative stress and hypoxia-inducible factor-1α in a dose-dependent manner. Folic acid targeted the activation of Janus kinase 2, downregulated the phosphorylation of signal transducer and activator of transcription 3, and decreased the expression of nuclear factor-κB p65 protein in cells. However, the absence of folic acid did not make cells more vulnerable under hypoxic conditions. In conclusion, folic acid efficiently inhibited the inflammatory response of THP-1 cells under hypoxic conditions by inhibiting reactive oxygen species production and the Janus kinase 2/signal transducer and activator of transcription 3 signaling pathway. Our study supports a basis for treatment with folic acid for chronic inflammation, which correlated with hypoxia.

Folic acid attenuates cobalt chloride-induced PGE2 production in HUVECs via the NO/HIF-1alpha/COX-2 pathway.

Prostaglandin E2 (PGE2), an important lipid inflammatory mediator involved in the progression of vascular diseases, can be induced by hypoxia in many cell types. While folic acid has been shown to protect against inflammation in THP-1 cells during hypoxia and hypoxia-induced endothelial cell injury, whether it might do so by attenuating PGE2 production remains unclear. To investigate this we constructed a hypoxia-induced injury model by treating human umbilical vein endothelial cells (HUVECs) with cobalt chloride (CoCl2), which mimics the effects of hypoxia. In CoCl2-treated HUVECs, folic acid significantly attenuated PGE2 production and increased vasoprotective nitric oxide (NO) content. Folic acid also decreased cyclooxygenase-2 (COX-2) and hypoxia-inducible factor 1-alpha (HIF-1α) expression and altered endothelial nitric oxide synthase (eNOS) signaling by increasing p-eNOS(Ser1177) and decreasing p-eNOS(Thr495) in a dose-dependent manner. Further investigation of the pathway demonstrated that treatment with 2-Methoxyestradiol (2-MeOE2) and celecoxib both decreased CoCl2-induced COX-2 expression but only 2-MeOE2 decreased HIF-1α expression. The ability of folic acid to down-regulate HIF-1α and COX-2 protein levels was dramatically abrogated by L-NAME treatment, which also decreased eNOS mRNA and NO production. The NO donor sodium nitroprusside also dose-dependently down-regulated HIF-1α and COX-2 protein levels. Overall, these findings suggest a novel application for folic acid in attenuating CoCl2-induced PGE2 production in HUVECs via regulation of the NO/HIF-1α/COX-2 pathway.

Folic Acid Represses Hypoxia-Induced Inflammation in THP-1 Cells through Inhibition of the PI3K/Akt/HIF-1α Pathway.

Though hypoxia has been implicated as a cause of inflammation, the underlying mechanism is not well understood. Folic acid has been shown to provide protection against oxidative stress and inflammation in patients with cardiovascular disease and various models approximating insult to tissue via inflammation. It has been reported that hypoxia-induced inflammation is associated with oxidative stress, upregulation of hypoxia-inducible factor 1-alpha (HIF-1α), and production of pro-inflammatory molecules. Whether folic acid protects human monocytic cells (THP-1 cells) against hypoxia-induced damage, however, remains unknown. We used THP-1 cells to establish a hypoxia-induced cellular injury model. Pretreating THP-1 cells with folic acid attenuated hypoxia-induced inflammatory responses, including a decrease in protein and mRNA levels of interleukin (IL)-1ß and tumor necrosis factor-alpha (TNF-α), coupled with increased levels of IL-10. Folic acid also reduced hypoxia-induced Akt phosphorylation and decreased nuclear accumulation of HIF-1α protein. Both LY294002 (a selective inhibitor of phosphatidyl inositol-3 kinase, PI3K) and KC7F2 (a HIF-1α inhibitor) reduced levels of hypoxia-induced inflammatory cytokines. We also found that insulin (an Akt activator) and dimethyloxallyl glycine (DMOG, a HIF-1α activator) induced over-expression of inflammatory cytokines, which could be blocked by folic acid. Taken together, these findings demonstrate how folic acid attenuates the hypoxia-induced inflammatory responses of THP-1 cells through inhibition of the PI3K/Akt/HIF-1α pathway.