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CpG-Oligodeoxynucleotides Alleviate Tert-Butyl Hydroperoxide-Induced Macrophage Apoptosis by Regulating Mitochondrial
Yibai Qu1, Chunxiu Yang1, Xueyang Li1
1Guangdong Provincial Key Laboratory of Proteomics, State Key Laboratory of Organ Failure Research, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China.
Abstract:
Oxidative stress and mitochondrial dysfunction are related to disease pathogenesis. Oligodeoxynucleotide containing CpG motifs (CpG ODN) demonstrate possibilities for immunotherapy applications. The aim of the present work is to explore the underlying mechanism of the cytoprotective function of CpG ODN by employing the oxidative stress modulation in immune cells. We used the imaging flow cytometry to demonstrate that tert-butyl hydroperoxide (t-BHP) induces mitochondrial-mediated apoptosis and ROS production in RAW264.7 cells. After pretreatment with CpG ODN, the percentage of apoptotic cells and ROS production was both markedly reduced. The decrease in mitochondrial membrane potential (MMP) induced by t-BHP was partially reversed by CpG ODN. The t-BHP induced upregulation of the expression of apoptosis-related proteins (cleaved-caspase 3, cleaved-caspase 9, cleaved-PARP, and bax) was notably decreased in the presence of CpG ODN. Furthermore, we found that CpG ODN enhanced phosphorylation of ERK1/2 and Akt to inhibit ROS production. In conclusion, the protective effect of CpG ODN in mitigation of t-BHP-induced apoptosis is dependent on the reduction of ROS.
Insights
CpG Oligodeoxynucleotides (CpG ODN) protect immune cells from oxidative stress by reducing reactive oxygen species (ROS) and apoptosis. This immunotherapy approach shows promise in mitigating disease pathogenesis.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Oxidative stress and mitochondrial dysfunction are implicated in disease pathogenesis.
- Oligodeoxynucleotides containing CpG motifs (CpG ODN) show potential for immunotherapy.
- Understanding the cytoprotective mechanisms of CpG ODN is crucial.
Purpose of the Study:
- To investigate the underlying mechanism of CpG ODN's cytoprotective function.
- To explore the modulation of oxidative stress in immune cells by CpG ODN.
- To assess the impact of CpG ODN on apoptosis and reactive oxygen species (ROS) production.
Main Methods:
- Utilized imaging flow cytometry to analyze RAW264.7 cells.
- Induced oxidative stress using tert-butyl hydroperoxide (t-BHP).
- Assessed apoptosis, ROS production, mitochondrial membrane potential (MMP), and protein expression.
Main Results:
- t-BHP induced apoptosis and ROS production, which were reduced by CpG ODN pretreatment.
- CpG ODN partially reversed t-BHP-induced decrease in MMP.
- CpG ODN decreased the expression of apoptosis-related proteins and enhanced ERK1/2 and Akt phosphorylation.
Conclusions:
- CpG ODN exhibits a cytoprotective effect against t-BHP-induced apoptosis in immune cells.
- This protective effect is mediated by the reduction of ROS production.
- CpG ODN's mechanism involves modulating oxidative stress pathways and enhancing cell survival signaling.
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