2.6 Animal experiments
Female C57BL/6 mice (8-10 weeks, 20-22g) and Nrf2 KO mice were purchased from Vital River (Beijing, China) and Jackson Laboratory (Bar, Harbor, USA), respectively. After one week of acclimatization, CI-AKI was established in female C57BL/6 mice. The experimental animals were divided into five groups as follows (n=10/group): control (0.1% DMSO in normal saline), cisplatin (20 mg/kg), cisplatin (20 mg/kg)+leonurine (50 mg/kg and 100 mg/kg) and leonurine only (100 mg/kg dissolved in 0.1% DMSO). After fasting, the mice in the cisplatin (20 mg/kg) + leonurine group were treated with 50 mg/kg and 100 mg/kg leonurine by gavage and intraperitoneally injected with cisplatin (20 mg/kg) one hour later to induce acute renal injury. Mice in the control, cisplatin and leonurine groups were treated with 0.1% DMSO in normal saline, cisplatin (20 mg/kg) and leonurine (100 mg/kg dissolved in 0.1% DMSO), respectively. On the second and third days, the mice in the cisplatin (20 mg/kg)+leonurine and leonurine only groups were administered leonurine by gavage once more. On the fourth day, all the mice were sacrificed, and kidney and blood samples were collected for further experiments. The mice were housed in a controlled living environment and provided adequate amounts of food and water.