Objective: To explore the mechanism of ischemia reperfusion injury (IRI) and the protective effect of Yisheng injection on endothelial cells (ECs).
Methods: Human umbilical veins endothelial cell line ECV304 was cultured in RPMI medium 1640 without glucose under 100% N2 for one and a half hours, and then in RPMI medium 1640 with glucose under normal conditions for 5 hours to mimic ECs' IRI posttransplantaion. A pure nature medicine, Yisheng injection was added into the medium in experimental group. Intracellular calcium and mitochondrial were shown through flourescent staining. The activities of succinic dehydrogenase (SDH) and lactic dehydrogenase (LDH) were detected by cytochemical staining. The concentrations of superoxide dismutase (SOD), malonaldehyde (MDA) and nitrous oxide (NO) in culture medium were determined.
Results: After treatment with anoxia-reoxygenation, the ECs' morphologic changes were observed, and intracellular calcium concentration, LDH activity, MDA concentration became higher. Meanwhile, mitochondrial membrane potential, concentrations of SOD and NO were lower. Yisheng injection made these changes disappear or become less.
Conclusion: Anoxia-reoxygenation induces lipid peroxidation, calcium superrcharge in ECs, and damage to mitochondrial function. Yisheng injection can reverse these changes, thus protecting the endothelial cells.