Failure to confirm a sodium-glucose cotransporter 2 inhibitor-induced hematopoietic effect in non-diabetic rats with renal anemia

Abstract

Aims/introduction: Clinical studies have shown that treatment with inhibitors of sodium-glucose cotransporter 2 (SGLT2) significantly increases the hematocrit in patients with type 2 diabetes. To investigate whether SGLT2 inhibitors directly promote erythropoietin production independently on blood glucose reduction, the hematopoietic effect of the specific SGLT2 inhibitor, luseogliflozin, was examined in non-diabetic rats with renal anemia.

Materials and methods: Renal anemia was induced by treatment with adenine (200 or 600 mg/kg/day, orally for 10 days) in non-diabetic Wistar-Kyoto or Wistar rats, respectively. Luseogliflozin (10 mg/kg bodyweight) or vehicle (0.5% carboxymethyl cellulose) was then administered for 6 weeks. The hematocrit and the hemoglobin (Hb), blood urea nitrogen, plasma creatinine, and plasma erythropoietin levels were monitored.

Results: Treatment with adenine decreased the hematocrit and the Hb level, which were associated with increases in the blood urea nitrogen and plasma creatinine levels. In Wistar-Kyoto rats treated with 200 mg/kg/day adenine, administration of luseogliflozin induced glycosuria, but did not change the blood urea nitrogen, plasma creatinine levels, hematocrit, Hb or plasma erythropoietin levels. Similarly, luseogliflozin treatment failed to change the hematocrit or the Hb levels in Wistar rats with renal anemia induced by 600 mg/kg/day of adenine. Plasma erythropoietin concentrations were also not different between luseogliflozin- and vehicle-treated rats. Similarly, in human erythropoietin-producing cells derived from pluripotent stem cells, luseogliflozin treatment did not change the erythropoietin level in the medium.

Conclusions: These data suggest that SGLT2 inhibitor fails to exert hematopoietic effects in non-diabetic conditions.

Keywords: Erythropoietin; Renal anemia; Sodium-glucose cotransporter 2 inhibitor.

Figure 1

Effect of luseogliflozin on the hematocrit and hemoglobin, and plasma erythropoietin levels in non‐diabetic Wistar–Kyoto rats with renal anemia. Wistar–Kyoto rats were treated with adenine (200 mg/kg/day) for 10 days to induce mild renal anemia. (a–b) the hematocrits (a) and hemoglobin levels (b) at 0, 2 and 6 weeks after luseogliflozin treatment. (c) The plasma erythropoietin levels before and after luseogliflozin treatment in adenine (200 mg/kg/day)‐treated Wistar–Kyoto rats. *P < 0.05, adenine + vehicle vs control.

Figure 2

Effect of luseogliflozin on tubulointerstitial injury in non‐diabetic Wistar–Kyoto rats with renal anemia. Interstitial fibrosis was evaluated by a semiquantitative analysis of azan staining. Scale bar, 100 µm. *P < 0.05, adenine + vehicle versus control.

Figure 3

Effect of luseogliflozin on the hematocrit and hemoglobin, and plasma erythropoietin levels in non‐diabetic Wistar rats with renal anemia. Wistar rats were treated with adenine (600 mg/kg/day) for 10 days to induce renal anemia. The (a) hematocrits and (b) hemoglobin levels at 0, 2 and 6 weeks after luseogliflozin treatment. (c) The plasma erythropoietin levels before and after luseogliflozin treatment in adenine (600 mg/kg/day)‐treated Wistar rats. *P < 0.05, adenine + vehicle versus control.

Figure 4

Effect of luseogliflozin on tubulointerstitial injury in non‐diabetic Wistar rats with renal anemia. Interstitial fibrosis was evaluated by a semiquantitative analysis of azan staining. Scale bar, 100 µm. *P < 0.05, adenine + vehicle versus control.

Figure 5

Effect of luseogliflozin (luseo) on erythropoietin production in human induced pluripotent stem cell‐derived erythropoietin‐producing cells. (a) Erythropoietin levels in the culture media after treatment with vehicle, luseogliflozin (100 and 500 nmol/L), or FG‐4592 (50 µmol/L) in human induced pluripotent stem cell‐derived erythropoietin‐producing cells. (b) Effects of luseogliflozin (100 nmol/L) on erythropoietin levels in the culture medium under 25 mmol/L or 50 mmol/L glucose concentrations in human induced pluripotent stem cell‐derived erythropoietin‐producing cells. *P < 0.05, FG‐4592 versus control.