Acetazolamide Suppresses Multi-Drug Resistance-Related Protein 1 and P-Glycoprotein Expression by Inhibiting Aquaporins Expression in a Mesial Temporal Epilepsy Rat Model

Abstract

BACKGROUND Mesial temporal epilepsy (MTLE) is the most common type of focal epilepsy in adults, and is often drug-resistant. This study investigated the effects of aquaporins (AQP) inhibitor on multi-drug-resistant protein expression in an MTLE rat model. MATERIAL AND METHODS The MTLE rat model was established by injecting pilocarpine into rats. The MTLE rats were divided into an MTLE-6 h group, an MTLE-12 h group, and an MTLE-24 h group, together with a normal saline group (NS), to examine the AQP4 expression by using Western blot assay and immunohistochemistry assay. The other 18 MTLE model rats were used to observe the effects of the AQP4 inhibitor, acetazolamide, on the multi-drug-resistant protein 1 (MRP1) and P-glycoprotein (Pgp) by using Western blot and immunohistochemistry assays, respectively. RESULTS AQP4 expression was enhanced in hippocampal tissues of MTLE model rats compared to NS rats (P<0.05). More positively stained AQP4 was discovered in hippocampal tissues of MTLE model rats. AQP4 inhibitor significantly decreased multi-drug-resistant protein MRP1 and Pgp expression in the AQP4 inhibitor Interfere group and the AQP4 inhibitor Therapy group compared to the TMLE model group (P<0.05). CONCLUSIONS The present findings confirm that the AQP4 inhibitor, acetazolamide, effectively inhibits the multi-drug-resistant protein, MRP1, and Pgp, in the MTLE rat model.

Figure 1

Observation for the AQP4 expression in MTLE model rats at 6 h, 12 h, and 24 h after the seizure. (A) Western blot bands for AQP4 expression in each group. (B) Statistical analysis for AQP4 expression in CA1 region. (C) Statistical analysis for AQP4 expression in CA3 region. (D) Statistical analysis for AQP4 expression in DG region. * P<0.05 and ** P<0.01 represent the AQP4 levels in MTLE model rats at 12 h and 24 h compared to the NS group.

Figure 2

AQP4 expression examined by using immunohistochemistry assay in MTLE model rats at 6 h, 12 h, and 24 h after the seizure. (A) Immunohistochemistry assay images for AQP4 staining. (B) Statistical analysis for AQP4 staining in CA1 region. (C) Statistical analysis for AQP4 staining in CA3 region. (D) Statistical analysis for AQP4 staining in DG region. * P<0.05 and ** P<0.01 represent the AQP4 staining in MTLE model rats at 6 h, 12 h, and 24 h compared to the NS group.

Figure 3

Multi-drug-resistant protein, MRP1 and Pgp, observation in AQP4 inhibitor-treated MTLE rats. (A) Western blot bands for the MRP1 and Pgp expression in each group. (B) Statistical analysis for MRP1 expression. (C) Statistical analysis for Pgp expression. * P<0.05 and ** P<0.01 represent the MRP1 or Pgp expression in the Interfere group or Therapy group compared to the MTLE rat model.

Figure 4

Immunohistochemistry assay examination for MRP1 and Pgp expression in AQP4-treated MTLE rats. (A) Immunohistochemistry assay images for MRP1 and Pgp staining. (B) Statistical analysis for MRP1 staining. (C) Statistical analysis for Pgp staining. * P<0.05 and ** P<0.01 represent the MRP1 or Pgp staining in the Interfere group or Therapy group compared to the MTLE rat model.