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Increased level of free-circulating MtDNA in maintenance hemodialysis patients: Possible role in systemic
Xiao-Yi Zhong1, Yi Guo2, Zhen Fan3
1Department of Nephrology, Clinical Medical College and The First Affiliated Hospital of Chengdu Medical College, Chengdu, China.
Background:
Mitochondrial DNA (MtDNA) exposed to the extracellular space due to cell death and stress has immunostimulatory properties. However, the clinical significance of circulating MtDNA in maintenance hemodialysis (MHD) patients and the precise mechanism of its emergence have yet to be investigated.
Methods:
This cross-sectional study consisted of 52 MHD patients and 32 age- and sex-matched healthy controls. MHD patients were further categorized into high and low circulating cell-free MtDNA (ccf-MtDNA) groups based on the median value. Copy number of MtDNA was quantified using TaqMan-based qPCR. Plasma cytokines were measured using ELISA kits. Reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm) in peripheral blood mononuclear cells (PBMCs) were detected using DCFH-DA or JC-1 staining.
Results:
The copy numbers of ccf-MtDNA in patients with MHD were higher than those in healthy controls, and these alterations were correlated with changes of cytokines TNF-α and IL-6. Adjusted model in multivariate analysis showed that the presence of anuria and longer dialysis vintage were independently associated with higher levels of ccf-MtDNA. Meanwhile, although not statistically significant, an inverse correlative trend between urinary MtDNA and ccf-MtDNA was observed in patients with residual urine. Afterward, using PBMCs as surrogates for mitochondria-rich cells, we found that patients in the high ccf-MtDNA group exhibited a significantly higher ROS production and lower Δψm in cells.
Conclusions:
Our data suggested that changes in ccf-MtDNA correlate with the degree of inflammatory status in MHD patients, and that the excessive MtDNA may be caused by mitochondrial dysfunction and reduced urinary MtDNA excretion.
Insights
Circulating cell-free mitochondrial DNA (ccf-MtDNA) is elevated in maintenance hemodialysis (MHD) patients, correlating with inflammation and potentially caused by mitochondrial dysfunction and reduced kidney excretion.
Area of Science:
- Nephrology
- Immunology
- Mitochondrial Biology
Background:
- Extracellular mitochondrial DNA (MtDNA) possesses immunostimulatory properties.
- The role and origin of circulating MtDNA in maintenance hemodialysis (MHD) patients remain unclear.
Purpose of the Study:
- To investigate the clinical significance and emergence mechanisms of circulating cell-free MtDNA (ccf-MtDNA) in MHD patients.
Main Methods:
- Cross-sectional study of 52 MHD patients and 32 healthy controls.
- Quantified ccf-MtDNA using qPCR, plasma cytokines via ELISA.
- Assessed reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm) in peripheral blood mononuclear cells (PBMCs).
Main Results:
- MHD patients exhibited higher ccf-MtDNA levels than controls, linked to elevated TNF-α and IL-6.
- Anuria and longer dialysis vintage independently predicted higher ccf-MtDNA.
- High ccf-MtDNA group showed increased ROS production and lower Δψm in PBMCs.
Conclusions:
- ccf-MtDNA levels correlate with inflammation in MHD patients.
- Mitochondrial dysfunction and decreased urinary MtDNA excretion may contribute to elevated ccf-MtDNA.
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