Kidney Week

Abstract: SA-PO160

Remote Ischemia Precondition Protects against Kidney Ischemia-Reperfusion Injury (IRI) through Apoptosis-Associated Vesicles Carrying MIF Protein

Session Information

• AKI: Metabolism and Cell Death
  October 26, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Acute Kidney Injury

• 103 AKI: Mechanisms

Authors

• Zhang, Nieke, Southeast University, Nanjing, Jiangsu, China

Nieke Zhang, MS

• Xia, Yi, Southeast University, Nanjing, Jiangsu, China

Yi Xia, MD

• Chen, Ming, Southeast University, Nanjing, Jiangsu, China

Ming Chen, PhD

• Zhang, Guangyuan, Southeast University, Nanjing, Jiangsu, China

Guangyuan Zhang, PhD

Background

Previous study showed remote ischemia precondition (rIPC) protected against kidney ischemia-reperfusion injury (IRI) through extracellular vesicles (EVs) delivering, however the mechanism remains unknown. It’s showed apoptosis-induced compensatory proliferation signaling related vesicles (ACPSVs) could transmit proliferation signal to surrounding cells. In this study, we aim to investigate the role of ACPSVs in renal IRI after rIPC and its underlying mechanism.

Methods

We established a bilateral hind limb IPC followed by unilateral renal IRI rat model and a serum-starved apoptotic cell model. ACPSVs were isolated from rat plasma or cellular supernatant using differential centrifugation, ACPSVs size was analyzed by electron microscopy and nanoparticle tracking analysis and their contents were analyzed by ELISA and Western blot. Observing the effects on renal IRI by HE staining, Ki67 and serum creatinine tests after allogeneic injection of ACPSVs or plasma post-IPC. Using CCK8 and flow cytometry analysis to investigate ACPSVs impact on cell proliferation and apoptosis after co-culture with HK2 cell. Applying GO and KEGG analysis to explore the potential downstream molecular mechanisms.

Results

A stable rat model was established. The characterization of ACPSVs was confirmed, and plasma or ACPSVs after rIPC can alleviate kidney damage, the protective effect was diminished when ACPSVs were removed from plasma. A tenfold increase in plasma macrophage migration inhibitory factor (MIF) expression after rIPC was observed, mainly concentrated in ACPSVs. A HeLa cell model was established, the optimal condition for ACPSVs generation was to culture cell in EBSS containing 0.1% BSA and 0.1g/L MgCl₂ for 24 hours. HK2 cells can uptake ACPSVs, with peak uptake at 12h post co-culture. EVs of approximately 460.6 nm in size were obtained, its ACPSV marker (CrkI) was confirmed. Compared to the sham group, apoptosis rates reduced and cell viability increased in the ACPSVs+HK2 group. Analysis of ACPSV contents highlighted the significance of MIF. GO and KEGG analysis showed that proteins contained in ACPSVs were enriched in molecular complexes related to macrophages and cell proliferation pathways.

Conclusion

rIPC not only exerts cytoprotective effects but also confers protection against renal IRI through ACPSVs carrying MIF protein.

Funding

• Government Support – Non-U.S.