HCQ Attenuates Hypoxia-Reoxygenation Injury

The HK-2 cells were plated in 35 mm dishes at a density of 10⁶ cells/ dish and incubated until they reached approximately 90% confluence for experiment. The cells in hypoxia/ reoxygenation (H/R) group were cultured for 12 h under hypoxic conditions (1% O₂, 94% N₂, and 5% CO₂) in medium without nutrients (glucose-free, serum-free) to induce hypoxic injury. After hypoxic treatment, the cells were transferred back to regular culture medium with oxygen for 2 h for reoxygenation. Control cells were incubated in complete culture medium in a regular incubator (5% CO₂ and 95% air). The cells in HCQ-treated groups were pretreated with HCQ (0.5, 2.5, and 5 μmol/L) for 12 h before H/R operation.