Nephroprotective Efficacy of Echinops spinosus against a Glycerol-Induced Acute Kidney Injury Model

Abstract

Nephroprotection or renal rescue is to revive and restore kidney function after damage, with no need for further dialysis. During acute kidney injury (AKI), sudden and recent reductions in kidney functions occur. Causes are multiple, and prompt intervention can be critical to diminish or prevent morbidity. Echinops spinosus (ES) is a curative plant with proven pharmacological and biological effects including anti-inflammatory, antioxidant, and antibacterial competencies. The principal goal of this research is to scrutinize the nephroprotective features of E. spinosa extract (ESE) against glycerol-induced AKI. Male Wistar albino rats were equally divided into five separated groups: negative control rats (vehicle-injected), ESE control rats (ESE-treated rats), positive control rats, glycerol-induced AKI-model rats (single IM injection of 50% glycerol), and 2 groups of diseased rats but pretreated with different concentrations of ESE for 7 days (ESE₁₅₀ + AKI rats and ESE₂₅₀ + AKI rats). Kidney tissues were collected and used for histopathology analysis. The relative kidney weight percentage was assessed. ESE effects were investigated via scanning several biomarkers, such as serum urea and creatinine, as kidney function biomarkers. Lactate dehydrogenase (LDH) and creatine kinase (CK) activities were examined as rhabdomyolysis (RM) indicators. Kidney injury molecule-1 (Kim-1) and neutrophil gelatinase-associated lipocalin (NGAL) were also examined to investigate kidney injury. Enzymatic and nonenzymatic oxidative stress markers were analyzed, namely, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), malondialdehyde (MDA), nitric oxide (NO), and reduced glutathione GSH. Proinflammatory cytokine [tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β)] and the renal proapoptotic protein (Bax) and antiapoptotic protein (Bcl-2) levels were evaluated. Statistical analysis for the resulting data revealed that ESE pretreatment turned AKI-induced biological antioxidant levels to an extent comparable to normal results. Furthermore, ESE decreased kidney function markers and RM-related biomarkers (LDH, CK, Kim-1, and NGAL) compared to those in untreated AKI-model rats. ESE treatment dropped the apoptotic renal Bax levels, enhanced antiapoptotic Bcl-2 manufacture, and disallowed the release of IL-1β and TNF-α. This study revealed the protective effect of ESE as therapeutic medicine against AKI-encouraged oxidative stress, inflammation, and apoptosis. It can be effectively used as adjuvant therapy, helping in renal rescue, and for kidney healing in cases with risk factors of AKI.

Figure 1

GC–MS chromatogram of active ingredients of E. spinosus.

Figure 2

Effects of E. spinosus extract (ESE) on kidney weight and relative kidney weight (RKW %) in glycerol-induced acute kidney injury. Results are presented as mean ± SD (n = 7); ^ψ and ^Δ represent significant changes at p < 0.05 between the untreated control and AKI groups, respectively.

Figure 3

Effects of E. spinosus extract (ESE) on kidney function (urea and creatinine), rhabdomyolysis (RM)-related (LDH and CK) markers, kidney injury molecule-1 (Kim-1), and neutrophil gelatinase-associated lipocalin (NGAL) in glycerol-induced acute kidney injury. Results are presented as mean ± SD (n = 7); ^ψ and ^Δ represent significant changes at p < 0.05 between the untreated control and AKI groups, respectively.

Figure 4

Effects of E. spinosus extract (ESE) on the renal levels of oxidative stress biomarkers [malondialdehyde (MDA), nitric oxide (NO), and total oxidant status (TOS)] and enzymatic and nonenzymatic molecules [glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR)] in glycerol-induced acute kidney injury. Results are presented as mean ± SD (n = 7); ^ψ and ^Δ represent significant changes at p < 0.05 between the untreated control and AKI groups, respectively.

Figure 5

Effects of E. spinosus extract (ESE) on the levels of proinflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β)] in glycerol-induced acute kidney injury. Results are presented as mean ± SD (n = 7); ^ψ and ^Δ represent significant changes at p < 0.05 between the untreated control and AKI groups, respectively.

Figure 6

Effects of E. spinosus extract (ESE) on the levels of apoptotic proteins [Bcl-2-associated X-protein (Bax), B-cell lymphoma 2 (Bcl-2), and Bcl-2/Bax ratio] in glycerol-induced acute kidney injury. Results are presented as mean ± SD (n = 7); ^ψ and ^Δ represent significant changes at p < 0.05 between the untreated control and AKI groups, respectively.

Figure 7

Effects of E. spinosus extract (ESE) on the histopathology of renal tissue in glycerol-induced acute kidney injury. (a) Control showing normal tissue architecture of the renal cortex, (b) ESE 250 mg showing the cortex area with normal structure and healthy glomeruli, (c) AKI showing severe tissue damage as evidenced by glomerular degeneration (white star), tubular dilatation and desquamation (blue arrow), hemorrhage (white arrow), vacuolation, necrosis, and debris buildup in the tubular lumina (green arrow), (d) ESE₁₅₀ + AKI showing some amelioration of the renal histological abnormalities, (e) ESE₂₅₀ + AKI showing a marked improvement in the renal tissue appearance, and (f) semiquantitative scoring of renal lesions. Hematoxylin and eosin (H&E). Scale bar = 100 μm. Results are presented as mean ± SD (12 fields per kidney, 2 kidneys per animal, and 3 animals per group were used in the analyses); ^ψ and ^Δ represent significant changes at p < 0.05 between the untreated control and AKI groups, respectively.