A total of 72 male C57BL/6 mice were purchased from Slyke jingda Biotechnology Company. They were randomly divided into five groups: Control group (n = 8), Cisplatin (20 mg/kg dissolved in saline) only group (n = 16), Cisplatin + paricalcitol (0.2 ug/kg dissolved in sterile water for injection and 20% propylene glycol) group (n = 16), Cisplatin + DMSO group (n = 16), Cisplatin + Fer-1 (5 mg/kg dissolved in DMSO) group (n = 16), were administered intraperitoneally. Cisplatin was injected once to mice, while Fer-1 was injected once an hour before cisplatin, and paricalcitol was injected once daily for five consecutive days before cisplatin. Each eight mice were sacrificed at 48 h and 72 h, respectively after cisplatin injection, and eight mice in the control group were sacrificed together with mice at 72 h.

Male C57BL/6 mice (aged 6-8 weeks and weighing 22-25g) were obtained from the Experimental Animal Center, Sichuan Provincial Peoples Hospital, and were fed a standard laboratory diet. LPS and ISL were dissolved in normal saline and 0.5% Tween-20/saline, respectively. AKI mice were developed by intraperitoneal (i.p.) LPS injection. A total of 30 mice were randomly divided into six groups (n = 5): control, ISL, Fer, LPS, LPS plus ISL, and LPS plus Fer. An intraperitoneal injection of LPS (10 mg/kg) was made to induce septic AKI. ISL was administered via gavage at 50 mg/kg 30 min before LPS injection. Mice were dosed intraperitoneally with Fer (Ferrostatin-1, SML0583, Sigma-Aldrich, St. Louis, MO) at 5 mg/kg. Mice were sacrificed by cervical dislocation 8 h after LPS injection. Kidney tissue and serum samples were collected concurrently.

Mice were fasted for 12 h and anesthetized (1% pentobarbital sodium, i.p.) before surgery. Bilateral renal pedicles were clamped for 30 min, then remove the arterial clamps. The sham groups were treated in the same way, except for the clamping of the renal pedicle. Blood samples were collected 24 h after reperfusion, mice were killed, and kidney were collected for follow-up experiments. Fedratinib (5 mg/kg body weight) was injected (i.p.) into mice 24 h once in advance before surgery.

The genetic background of embryonic stem cells and the Flp mice used in this experiment was C57BL/6. Mice were randomly separated into experimental groups and control groups. (1) Bilateral IRI: mice (male, 8-10 weeks old) on the lgmnKO background or littermate control mice were anesthetized by an intraperitoneal (i.p.) injection of chloral hydrate and placed on a warm pad to retain their body temperature. A bilateral flank incision was made, both sides of the renal vessels were occluded with clamps for 40 min followed by removing the clamps to induce blood reperfusion. The same procedure was performed in the control group without vessel clamping. (2) Nephrotoxic folic acid-induced AKI: mice (female, 12-14 weeks old) received a single i.p. injection of folic acid at 250 mg/kg in 0.3 mol/L sodium bicarbonate or the vehicle. For therapeutic experiments, RR-11a was freshly dissolved in saline. Mice were administered an i.p. injection of 20 mg/kg RR-11a or the vehicle before ischemia.

Male C57BL/6 mice (8-10 weeks of age, weight 20-25 g) were purchased from Experimental Animal Center of the Fourth Military Medical University (Xi'an, China) and bred in an experimental animal room of SPF grade. They were randomly divided into four groups: control (equivalent saline containing 1% DMSO) group (n = 5), cisplatin (20 mg/kg dissolved in saline) only group (n = 7), cisplatin + polydatin (40 mg/kg dissolved in 1% DMSO) group (n = 7), and cisplatin+ Fer-1 (5 mg/kg dissolved in 1% DMSO) group (n = 7) were administered intraperitoneally. Mice were injected with cisplatin once; PD or Fer-1 was given 1 h before and 24 h after cisplatin. Animals were ethically sacrificed by dislocating their spines at 48 h after cisplatin injection, and whole blood and kidneys were collected for further analysis.

The C57BL/6 male mice (8-week-old, weighing approximately 20-25 g) were procured from Beijing Huafukang Bioscience Co. Inc. The mice were raised under the SPF condition. Cisplatin was injected into the mice intraperitoneally and only once at a dose of 30 mg/kg to induce AKI, while the control mice were injected with PBS. Intravenous administration of 10 mg/kg of agomir negative control (agomir NC) or agomir miR-214-3p (GenePharma Co. Ltd, Shanghai, China) was performed for the control group mice and model group mice, respectively. The ferroptosis inhibitor named Fer-1 (#S7243, Selleck Chemicals, Houston, TX, USA) was dissolved in DMSO and then diluted in 0.9% NaCl to prepare separate Fer-1 solutions each with a concentration of 0.2 mg/mL. Fer-1 was injected into the mice intraperitoneally, 1 h prior to injecting cisplatin, while the control mice received the injection of only 0.9% NaCl in 0.1% DMSO. Both experimental and control group mice were sacrificed at Day 1, 2, and 3, separately, post-cisplatin injection.

Male C57BL/6 mice (8-10 weeks; 20-25 g) were purchased from LINGCHANG BIOTECH (China). Mice were divided into four groups: (i) sham, (ii) I/R, (iii) I/R+entacapone, and (iv) I/R + Fer-1. Entacapone (15 mg/kg bodyweight) was dissolved in sodium carboxymethyl cellulose (0.5%) and administered (i.g.) to mice. Mice in the sham group were administered (i.g.) an equal volume of solvent. Fer-1 was dissolved in 5% dimethyl sulfoxide + 30% polyethylene glycol-400 + 60% saline and injected (i.p.). Mice were treated three times per day for 3 days in advance. Before I/R, mice were fasted for 12 h and anesthetized (1% pentobarbital sodium, i.p.). The abdomen was exposed and bilateral renal pedicles were clamped to induce renal I/R. After 25 min, the arterial clamps were removed. A body temperature of 37 was maintained throughout the procedure. The sham group underwent the same procedure except for clamping of the renal pedicle. Mice were killed 24 h after reperfusion, and kidney and blood samples were collected for experimentation.

Male C57BL/6 mice (8-10 weeks; 20-25 g) were purchased from LINGCHANG BIOTECH (China). Mice were divided into four groups: (i) sham, (ii) I/R, (iii) I/R+entacapone, and (iv) I/R + Fer-1. Entacapone (15 mg/kg bodyweight) was dissolved in sodium carboxymethyl cellulose (0.5%) and administered (i.g.) to mice. Mice in the sham group were administered (i.g.) an equal volume of solvent. Fer-1 was dissolved in 5% dimethyl sulfoxide + 30% polyethylene glycol-400 + 60% saline and injected (i.p.). Mice were treated three times per day for 3 days in advance. Before I/R, mice were fasted for 12 h and anesthetized (1% pentobarbital sodium, i.p.). The abdomen was exposed and bilateral renal pedicles were clamped to induce renal I/R. After 25 min, the arterial clamps were removed. A body temperature of 37 was maintained throughout the procedure. The sham group underwent the same procedure except for clamping of the renal pedicle. Mice were killed 24 h after reperfusion, and kidney and blood samples were collected for experimentation.

Six-week-old male Wistar rats (170-200 g) were obtained from Changsheng Biotechnology Co., Ltd. (Changchun, China), and all of them were fed under SPF-conditions. The rats were acclimatized to natural light/dark cycles at a controlled temperature of 22 + 2 with free access to food and water. The experiment was comprised of four groups: the C group (0.5% carboxymethyl cellulose sodium [CMC-Na], n = 6); the Dio group (dioscin-treated rats, n = 6); the CP group (cisplatin-treated mice, n = 6); and the Dio + CP group (dioscin plus cisplatin-treated rats, n = 6). Rats were gavaged with dioscin (60 mg/kg) for ten days, and cisplatin (10 mg/kg) was intraperitoneally injected once on the seventh day.

A total of 30 C57BL/6 male mice (8-10 weeks; 20-25 g body weight) were purchased from Chongqing Medical University (Chongqing, China). The mice were anesthetized with 50-60 mg/kg of pentobarbital sodium (cat. no. P3761; Sigma-Aldrich; Merck KGaA) by intraperitoneal injection; the skin at the surgical area was wiped with 70% alcohol. The incision was positioned at the left and right sides of the spine (0.5 cm), and the incision length was 1-1.5 cm along the back. The kidneys were subsequently pulled out from the incision to expose the renal pedicle. A microaneurysm clip was used to clamp the pedicle to block the blood flow to the kidney and induce renal ischemia. Complete ischemia was indicated by a change in the color of the kidney from red to dark purple within a few seconds. After 40 min of ischemia, the microaneurysm clips were released to allow each kidney to start reperfusion, which was indicated by the change of the kidney color to red.

C57BL/6 mice (8-12 week) mice purchased from Cavens (Changzhou, China) were divided into four groups: control (saline), CDDP only (CP, 20 mg/kg; MCE, United States), CP (20 mg/kg) + ADAMTS13 (0.1 nmol/kg) and CP (20 mg/kg) + ADAMTS13 (0.3 nmol/kg). Each group contained 5 mice. 0.1 and 0.3 nmol/kg rhADAMTS13 were injected into the caudal vein daily for 3 days after surgery.

Thirty-two male C57BL/6 mice (20 ± 2) g (Beijing Weitonglihua Experimental Animal Technology Co., Ltd.) were bred in individually ventilated cages (IVC) at SPF conditions, kept on a 12 h light/dark cycle, relative humidity conditions (40-70%) and controlled temperature (24 ± 2 ). After one-week acclimation mice were divided randomly into four groups: control group, cisplatin group (20 mg/kg cisplatin dissolved in saline), cisplatin + Fer-1 group (5 mg/kg Fer-1 dissolved in DMSO), and cisplatin + gap27 group (35 ug/kg gap27 dissolved in DMSO). There were eight animals in each group and 20 mg/kg cisplatin was given to each animal once by intraperitoneal injection except mice in the control group. Fer-1 and gap27 was administered 1 h before the injection of cisplatin.

Gene knockout (Rev-erb-a-/-,Rev-erb-b-/-and icDKO) mice and wild-type littermates were treated with folic acid (i.p., 100 mg/kg, once daily for seven consecutive days) at ZT6 or ZT18 to induce acute kidney injury.

Gene knockout (Rev-erb-a-/-,Rev-erb-b-/-and icDKO) mice and wild-type littermates were treated with folic acid (i.p., 100 mg/kg, once daily for seven consecutive days) at ZT6 or ZT18 to induce acute kidney injury.

Thirty-two male C57BL/6 mice (20 ± 2) g (Beijing Weitonglihua Experimental Animal Technology Co., Ltd.) were bred in individually ventilated cages (IVC) at SPF conditions. After one-week acclimation mice were divided randomly into four groups: control group, cisplatin group (20 mg/kg cisplatin dissolved in saline), cisplatin + Fer-1 group (5 mg/kg Fer-1 dissolved in DMSO), and cisplatin + gap27 group (35 ug/kg gap27 dissolved in DMSO). There were eight animals in each group and 20 mg/kg cisplatin was given to each animal once by intraperitoneal injection except mice in the control group. Fer-1 and gap27 was administered 1 h before the injection of cisplatin.

All experiments were performed according to the protocols approved by the Animal Care Committee of Wonkwang University. AKI was induced by a single intraperitoneal injection of cisplatin (10 mg/kg). One h before cisplatin injection, mice in the loganin group received 1, 10, or 20 mg/kg of loganin orally, and mice in the U0126 group received 10 mg/kg of U0126 intraperitoneally. They were sacrificed at 72 h after cisplatin injection, and their blood and kidneys were collected.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

Male C57BL/6 mice (8-10 weeks, 20-25 g) were purchased from HuaFuKang Company (Beijing, China). After 1 week of adaptation to the housing conditions, mice were intraperitoneally injected with folic acid (250 mg/kg) to induce acute kidney injury. An injection of sodium bicarbonate (0.3-M NaHCO3, the vehicle used for folic acid treatment) alone was used as a negative control. Nuciferine (30 mg/kg) was dissolved in water, sonicated, and then immediately administered to mice intragastrically. The sham control mice were treated with nuciferine but not folic acid.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

C57BL/6N male mice (CLEA Japan), aged 8-9 weeks, were used. AKI was induced by intraperitoneal injection of cisplatin solution (16 or 17 mg/kg as indicated; Nichi-Iko Pharmaceutical). Mice were orally treated with water only, promethazine (20 mg/kg in water), or rifampicin (20 mg/kg in 0.5% methylcellulose) every 12 hours for 4 days starting 30 minutes before the cisplatin injection, or orally treated with promethazine (20 mg/kg) in the following groups: (1) no promethazine, (2) pretreatment 30 minutes before cisplatin injection, (3) treatment from 30 minutes before injection to 24 hours after injection, (4) treatment from 24 to 96 hours after injection, and (5) treatment every 12 hours from 30 minutes before injection to 96 hours after injection.

Homozygous Rheb1 floxed mice (C57BL/6J background) were kindly provided by Dr. Xiao. To induce AKI in mice, Tubule-Rheb1-/-, Tubule-Tsc1+/- and their control littermates aged between 8 and 10 weeks were injected with a single dose of 20 mg/kg cisplatin (cat: P4394, Sigma-Aldrich, St. Louis, MO) intraperitoneally. Mice were sacrificed at day 1, 2 and 3 after cisplatin administration, and mice in AKI models died before being sacrificed were excluded.

For FA-induced nephropathy mouse models, 8-week-old male wild-type and Dpep1+/- or Chmp1a+/- mice were injected with FA (250 or 200 mg/kg, dissolved in 300 mM sodium bicarbonate) intraperitoneally and euthanized on day 7. For the cisplatin-induced injury model, 8-week-old male wild-type, Dpep1+/- or Chmp1a+/- mice were injected with cisplatin (25 or 20 mg/kg) intraperitoneally.

For FA-induced nephropathy mouse models, 8-week-old male wild-type and Dpep1+/- or Chmp1a+/- mice were injected with FA (250 or 200 mg/kg, dissolved in 300 mM sodium bicarbonate) intraperitoneally and euthanized on day 7. For the cisplatin-induced injury model, 8-week-old male wild-type, Dpep1+/- or Chmp1a+/- mice were injected with cisplatin (25 or 20 mg/kg) intraperitoneally.

To induce AKI in mice, Tubule-Rheb1-/-, Tubule-Tsc1+/-and their control littermates aged between 8 and 10 weeks were injected with a single dose of 20 mg/kg cisplatin (cat: P4394, Sigma-Aldrich, St. Louis, MO) intraperitoneally. Mice were sacrificed at day 1, 2 and 3 after cisplatin administration, and mice in AKI models died before being sacrificed were excluded.

Male C57BL/6 mice (aged 6-8 weeks and weighing 22-25g) were obtained from the Experimental Animal Center, Sichuan Provincial Peoples Hospital, and were fed a standard laboratory diet. LPS and ISL were dissolved in normal saline and 0.5% Tween-20/saline, respectively. AKI mice were developed by intraperitoneal (i.p.) LPS injection. A total of 30 mice were randomly divided into six groups (n = 5): control, ISL, Fer, LPS, LPS plus ISL, and LPS plus Fer. An intraperitoneal injection of LPS (10 mg/kg) was made to induce septic AKI. ISL was administered via gavage at 50 mg/kg 30 min before LPS injection. Mice were dosed intraperitoneally with Fer (Ferrostatin-1, SML0583, Sigma-Aldrich, St. Louis, MO) at 5 mg/kg. Mice were sacrificed by cervical dislocation 8 h after LPS injection. Kidney tissue and serum samples were collected concurrently.

All mice used in our in vivo studies were 8-week-old males of the C57BL/6J background. Kidneys were exposed via a midline abdominal incision and bilateral renal pedicle clamping for 35 min using microaneurysm clamps (Aesculap Inc., Center Valley, PA, USA). Throughout the surgical procedure, the mice were kept under isoflurane narcosis, and their body temperature was maintained at 36-37 by continuous monitoring using a temperature-controlled, self-regulated heating system (Fine Science Tools, Heidelberg, Germany). After clamps were removed, kidney reperfusion was confirmed visually before the abdomen was closed in two layers using standard 6-0 sutures. To maintain fluid balance, all mice were supplemented with 1 ml of prewarmed PBS administered intraperitoneally directly after surgery. After 48 h of reperfusion, the mice were sacrificed, blood samples were obtained by retrobulbar puncture, and kidneys were collected for analysis.

Six-week-old male Wistar rats (170-200 g) were obtained from Changsheng Biotechnology Co., Ltd. (Changchun, China), and all of them were fed under SPF-conditions. The rats were acclimatized to natural light/dark cycles at a controlled temperature of 22 + 2 with free access to food and water. The experiment was comprised of four groups: the C group (0.5% carboxymethyl cellulose sodium [CMC-Na], n = 6); the Dio group (dioscin-treated rats, n = 6); the CP group (cisplatin-treated mice, n = 6); and the Dio + CP group (dioscin plus cisplatin-treated rats, n = 6). Rats were gavaged with dioscin (60 mg/kg) for ten days, and cisplatin (10 mg/kg) was intraperitoneally injected once on the seventh day.

Gene knockout (Rev-erb-a-/-,Rev-erb-b-/-and icDKO) mice and wild-type littermates were treated with folic acid (i.p., 100 mg/kg, once daily for seven consecutive days) at ZT6 or ZT18 to induce acute kidney injury.

Gene knockout (Rev-erb-a-/-,Rev-erb-b-/-and icDKO) mice and wild-type littermates were treated with folic acid (i.p., 100 mg/kg, once daily for seven consecutive days) at ZT6 or ZT18 to induce acute kidney injury.