The Mitochondrial-Derived Peptide MOTS-c Attenuates Oxidative Stress Injury and the Inflammatory Response of H9c2 Cells Through the Nrf2/ARE and NF-κB Pathways

Abstract

Aim-Oxidative stress and the inflammatory response contribute to the progression of cardiovascular disease. The present study aimed to investigate whether the mitochondrial-derived peptide MOTS-c could alleviate H 2 O 2 -induced oxidative stress and inflammatory status in H9c2 cells through activation of nuclear factor erythroid 2-related Factor 2 (Nrf2)/antioxidative response element (ARE) and inhibition of the NF-jB pathway. Methods-Rat H9c2 cardiomyocytes were obtained, and 10, 20 or 50 lM MOTS-c was pretreated for 24 h before treatment with H 2 O 2. Then, the cell was treated with 100 lM H 2 O 2 for 1 h to induce oxidative stress. An inhibition model of sh-Nrf2 was constructed via a lentivirus expression system, and an activation model of NF-jB was achieved using phorbol 12-myristate-13-acetate (PMA). Cell viability was determined using a Cell Counting kit-8 assay. Relative measurement of relative protein and mRNA expression used western blotting and qRT-PCR, respectively. Intracellular reactive oxygen species (ROS) levels were detected using dichlorodihydrofluorescein diacetate, and malondialdehyde (MDA) and superoxide dismutase (SOD) levels were determined via commercial kits. The protein expression and distribution in the cells were visualized by immunofluorescence analysis. Enzyme-linked immunosorbent assay was used to detect the levels of inflammatory cytokines, including TNF-a, IL-6 and IL-1b. Results-We found that H 2 O 2 treatment significantly decreased cell viability and the level of SOD, increased the levels of ROS and MDA, and upregulated the expression of inflammatory cytokines, including TNF-a, IL-6 and IL-1b, in H9c2 cells. The expression levels of Nrf2, HO-1 and NQO-1 were significantly downregulated in the H 2 O 2 , while the phosphorylation of NF-jBp65 was promoted by H 2 O 2 . However, pretreatment with MOTS-c significantly reversed H 2 O 2 -induced damage in H9c2 cells. Moreover, both inhibition of the Nrf2/ARE pathway and activation of the NF-jB pathway significantly decreased the effects of MOTS-c, suggesting that MOTS-c might play a role in alleviating oxidative damage via the Nrf2/ARE and NF-jB pathways. Conclusions-Our investigation indicated that MOTS-c could protect against H 2 O 2 -induced inflammation and oxidative stress in H9c2 cells by inhibiting NF-jB and activating the Nrf2/ARE pathways.

Selenium Deficiency via the ROS/NLRP3/IL-1β Signaling Pathway Leads to Pyroptosis Injury in Pig Spleen

“…It can regulate related in ammatory factors through the MAPK signaling pathway. The content of p-Nrf2 is signi cantly reduced after H 2 O 2 treatment [30]. As Fig.…”

Heart Rate Variability Analysis Reveals a Non-monotonic Relationship between Humanin Concentration and Cardiac Autonomic Regulation

“…Given the important role of HN in lowering mitochondrial OS, our results could be interpreted as the impact of HN levels on autonomic modulation through its antioxidant properties. Future studies should investigate mitochondrial markers other than HN to gain insight into mitochondrial influence on the ANS through other MDPS such as MOTS-c, which potentially alleviates OS [22], and P66-Shc, an adaptor protein that promotes ROS production [23]. Furthermore, given that non-linear HRV measures showed the greatest differences, more non-linear measures could be explored in subsequent investigations to observe whether similar results are obtained.…”

Mitochondrial Dysfunction and Increased DNA Damage in Vascular Smooth Muscle Cells of Abdominal Aortic Aneurysm (AAA-SMC)

“…Activation of the ARE element in genes encoding detoxification enzymes by the transcription factor Nrf2 protects cells from oxidative stress-induced cell death. The Nrf2/ARE-pathway has been shown to be important for protection against H 2 O 2 -induced inflammation and oxidative stress in cardiomyocytes [ 46 ], and alterations in this cascade have been reported to be associated with neurodegenerative diseases [ 47 ]. Given the importance of ROS detoxification, it will be very interesting to investigate Nrf2 and other components of this pathway for their expression and function in vascular cells in connection with AAA.…”

“…Indeed, our data demonstrated increased O 2- levels in AAA-SMC. The fact that mitochondria-targeted MitoQ was able to reduce O 2- levels in both controls and AngII-challenged healthy VSMCs, but not in AAA-SMCs, suggests involvement of mitochondria in activation of the Nrf2/ARE cascade, as it was demonstrated in cardiomyocytes [ 46 ]. Interestingly, the level of another ROS, H 2 O 2 , was clearly higher in healthy VSMC cultures than in AAA-SMC after 24 h. Moreover, the amount of H 2 O 2 was significantly increased in MitoQ-treated cells after 24 h, again with a stronger effect in healthy VSMC than in AAA-SMC.…”