Effects of furosemide and tadalafil in both conventional and nanoforms against adenine-induced chronic renal failure in rats

Abstract

Background: Chronic renal failure (CRF) is a progressive loss of renal function that lead to reduced sodium filtration and inappropriate suppression of tubular reabsorption that ultimately leads to volume expansion. The aim of this study was to study the efficacy of furosemide and tadalafil nanoforms compared to conventional forms against adenine-induced CRF rat-model.

Methods: Addition of 0.75% adenine to the diet of rats for 4 weeks gained general acceptance as a model to study kidney damage as this intervention mimicked most of the structural and functional changes seen in human chronic kidney disease Urine analysis, histopathological changes and immunohistochemical expression of caspase-3 and interleukin-1 beta (IL-1β) in renal tissues were performed.

Results: Our results showed that the combination of tadalafil and furosemide using conventional and nanoparticle formulations had better renoprotective effect than individual drugs. This was demonstrated by improvement of urinary, serum and renal tissue markers as indicative of organ damage. This was also reflected on the reduction of tubular expression of kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL). Immunohistochemical studies showed that the deteriorated renal cellular changes indicated by increased expression of caspase-3 and IL-1β were greatly improved by the combined treatment particularly with the nanoforms.

Conclusions: The nanoforms of both furosemide and tadalafil had greater renopreventive effects compared with conventional forms against adenine-induced CRF in rats.

Keywords: Adenine–CRF in rats; Caspase-3; Furosemide; IL-1β; NGAL; Nanoparticles; Tadalafil.

Fig. 1

Zeta potential distribution of chitosan/alginate nanoparticles (A) and furosemide loading in chitosan/alginate nanoparticles (B) & zeta potential distribution of poly lactic-co-glycolic acid nanoparticles (C) and tadalafil loading in poly lactic-co-glycolic nanoparticles (D)

Fig. 2

Transmission electron microscopy photomicrographs depicting the spheroidal forms of furosemide nanoparticles (A) and the regular spherical shape of tadalafil nanoparticles (B) at different magnifications

Fig. 3

Effect of furosemide (F) and tadalafil (T) in their conventional, nanoparticle (N) forms and their combinations (o) on the levels of glucose, albumin and total ketone bodies in rat’s urine in adenine (A)-induced chronic renal failure. Data are the means ± SEM (n = 6). ****p < 0.0001 as compared to the adenine-treated group. ^ap < 0.0001 as compared with the control group. ^b p < 0.001 as compared with corresponding nanoparticle group (unpaired t test). ^c p < 0.05 as compared with corresponding nanoparticle group (unpaired t test)

Fig. 4

Effect of furosemide (F) and tadalafil (T) in their conventional, nanoparticle (N) forms and their combinations (o) on the urine osmolarity and specific gravity in rats in adenine (A)-induced chronic renal failure. Data are the means ± SEM (n = 6). ****p < 0.0001 as compared to the adenine group. ^ap < 0.0001 as compared with the control group. ^bp < 0.0001 as compared with corresponding nanoparticle group (unpaired t test)

Fig. 5

Effect of furosemide (F) and tadalafil (T) in their conventional, nanoparticle (N) forms and their combinations (o) on the levels of creatinine, urea and total protein in rats serum in adenine (A)-induced chronic renal failure. Data are the means ± SEM (n = 6). ****p < 0.0001, ***p < 0.001, **p < 0.01 and *p < 0.05 as compared to the adenine group. ^ap < 0.0001 as compared with the control group. ^bp < 0.05 as compared with corresponding nanoparticle group (unpaired t test)

Fig. 6

Effect of furosemide (F) and tadalafil (T) in their conventional, nanoparticle (N) forms and their combinations (o) on the levels of malondialdehyde, nitrite and reduced glutathione (GSH) in rat’s kidney tissue in adenine (A)-induced chronic renal failure. Data are the means ± SEM (n = 6). ****p < 0.0001, ***p < 0.001, **p < 0.01 and *p < 0.05 as compared to the adenine group. ^ap < 0.01 as compared with the control group. ^bp < 0.01as compared with corresponding nanoparticle group (unpaired t test). ^cp < 0.01 as compared with corresponding nanoparticle group (unpaired t test)

Fig. 7

Effect of furosemide (F) and tadalafil (T) in their conventional, nanoparticle (N) forms and their combinations (o) on the levels of kidney injury molecule 1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL) in rats kidney tissue in adenine (A)-induced chronic renal failure. Data are the means ± SEM (n = 6). ****p < 0.0001 as compared to the chronic group. ^ap < 0.0001 as compared with the control group. ^bp < 0.05 as compared with corresponding nanoparticle group (unpaired t test). ^cp < 0.05 as compared with corresponding nanoparticle group (unpaired t test). ^dp < 0.05 as compared with corresponding nanoparticle group (unpaired t test)

Fig. 8

Effect of furosemide and tadalafil in their conventional, nanoparticle forms and their combinations on the histopathological changes as represented in sections (A–H) and immunoreactivity of both IL-1β sections (I–P) and caspase-3 sections (Q–X) in rats kidney tissue in adenine-induced chronic renal failure (CRF)