TWEAK/Fn14 promotes oxidative stress through AMPK/PGC‑1α/MnSOD signaling pathway in endothelial cells

Abstract

Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) contributes to dysfunction of endothelial cells via its receptor, Fn14. However, its role in the production of reactive oxygen species (ROS), particularly mitochondrial ROS (mtROS) and the subsequent decrease in nitric oxide (NO) in endothelial cells remains unclear. In this study, the effect of TWEAK/Fn14 on generation of ROS, mtROS and NO in endothelial cells and its potential mechanism was investigated. Human umbilical vein endothelial cells (HUVECs) were treated with TWEAK with Fn14 small interfering (si)RNA or negative control RNA. It was demonstrated that TWEAK induced the production of ROS and mtROS in HUVECs, which were detected by fluorescent microscope, and flow cytometry. In addition, TWEAK decreased the generation of NO as indicated using the Nitric Oxide Assay kit. Furthermore, TWEAK aggravated mtDNA damage as measured by quantitative polymerase chain reaction analysis. Inhibition of Fn14 by Fn14 siRNA decreased TWEAK‑induced ROS and mtROS production, as well as mtDNA damage, while it increased the production of NO in endothelial cells. In addition, TWEAK inhibited the expression of active AMP‑activated protein kinase (AMPK) and its downstream protein peroxisome proliferator‑activated receptor‑γ coactivator-1α (PGC‑1α) and manganese superoxide dismutase (MnSOD). Notably, Fn14 siRNA enhanced the expression of the aforementioned proteins. Taken together, TWEAK/Fn14 contributes to endothelial dysfunction through modulation of ROS and mtROS. In addition, the underlying mechanism is implicated in the AMPK/PGC‑1α/MnSOD signaling pathway.

Short-Chain Fatty Acids and Their Association with Signalling Pathways in Inflammation, Glucose and Lipid Metabolism

“…Similar results were found by Kondo and Gao et al In their study, SCFA directly activated the AMPK signalling pathway by increasing the AMP/ATP ratio in the muscles and liver [13,119]. AMPK can affect many downstream signalling pathways, such as mTOR, PGC-1A and FOX03 [120][121][122]; however, whether these signalling pathways are related to glucose metabolism activated by SCFA remains to be further studied.…”

Tumor Necrosis Factor-Like Weak Inducer of Apoptosis (TWEAK) Enhances Activation of STAT3/NLRC4 Inflammasome Signaling Axis through PKCδ in Astrocytes: Implications for Parkinson’s Disease

“…Consistent with a previous report, 100 ng/mL TWEAK failed to elicit cell death in U373 human astrocytic cells ( Figure S1A) [38]. Thus, based on our cell viability studies showing a lack of toxicity, together with other reports [38,59,60] showing that 100 ng/mL TWEAK elicits a proinflammatory response in diverse cell culture models, we utilized this dosing regimen to investigate the TWEAK-induced astroglial activation response for our remaining studies. The U373 astrocytic cells were treated with 100 ng/mL TWEAK for the indicated durations (6, 12, 18, 24 h), and then ROS and mitochondrial (mito)ROS generation were determined by DCFDA and MitoSOX fluorescence plate reader assay, respectively.…”

Vildagliptin improves high glucose‐induced endothelial mitochondrial dysfunction via inhibiting mitochondrial fission

“…Mitochondrial DNA (mtDNA) damage in HUVECs was analysed by quantitative PCR as previously described . Briefly, DNA was extracted using a QIAamp DNA Micro kit (Qiagen, West Sussex, UK) and real‐time PCR reactions were performed on the Mastercycler EP realplex PCR System (Eppendorf, Wesseling‐Berzdorf, Germany) according to manufacturer's protocol.…”

“…Mitochondrial DNA (mtDNA) damage in HUVECs was analysed by quantitative PCR as previously described. 23 Briefly, DNA was…”