The protective effects of exogenous spermine on renal ischemia-reperfusion injury in rats

Abstract

Background: To investigate the protective effects of exogenous spermine on renal ischemia-reperfusion injury in rats.Methods: (I) Different doses of spermine were injected into rats to determine the safe dose on the kidneys. Kidney toxicity was assessed by hematoxylin and eosin (HE) staining of kidney tissue and enzyme-linked immunosorbent assay (ELISA) detection of neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule 1 (KIM-1) in the venous blood. (II) A rat model of renal ischemia-reperfusion injury was established. Different doses of spermine were injected into the rats through the tail vein 30 minutes before and 3 days after the establishment of the model. Blood samples and kidney tissues were collected and renal injury was assessed via HE staining of the renal tissue, detection of apoptosis using the TUNEL assay, and detection of NGAL and KIM-1 in blood samples using ELISA. (III) Human HK-2 renal tubular epithelial cells were cultured under hypoxia/reoxygenation conditions. To evaluate the protective effects of spermine, apoptosis was assessed by flow cytometry and TUNEL assay. The mechanisms underlying the effects of spermine were studied using Western blot analyses.Results: At spermine concentrations below 200 μM (2 mL/kg body weight), no significant damage to the kidney was observed by HE staining, and there was no significant difference in NGAL and KIM-1 levels between rats treated with spermine and control rats (P<0.05). At spermine doses below 200 μM, HE staining showed that the degree of renal ischemia-reperfusion injury was gradually alleviated with increasing doses of spermine. TUNEL assays demonstrated that spermine reduced the apoptosis of renal tissue, and increasing doses of spermine gradually decreased the levels of NGAL and KIM-1 in the blood compared with the control group (P<0.05). Western blot analysis revealed that spermine increased the expression of pro-caspase9, phosphorylated protein kinase B (p-Akt), hypoxia-inducible factor 1 alpha (HIF-1α), B cell lymphoma 2 (Bcl-2), and Bcl2 interacting protein 3 (BNIP3), and decreased the expression of cleaved caspase-3, Bax and cytochrome C compared to control cells.Conclusions: Exogenous spermine exerted a protective effect on renal ischemia-reperfusion injury in rats by inhibiting the apoptosis of renal tubular epithelial cells.

Thymoquinone and Oleuropein Combination Ameliorates Renal Ischemia-Reperfusion Injury by Attenuating Oxidative Stress in Rats

“…4b). These outcomes were following the previous reports [58][59][60][61] and further substantiate the renal injury caused by the IR surgery. Moreover, a graded pattern of morphological mutilation was seen in the pretreatment groups, which again indicates that TQ and OP have potent nephroprotective potential.…”

Metabolism of Polyamines and Kidney Disease: A Promising Therapeutic Target

“…Renal tubular repair may be completed after mild AKI, but incomplete or maladaptive repair usually occurs after severe or recurrent AKI, leading to the development of CKD [ 98 ]. Intraperitoneal injection of spermine protected against IRI-induced renal tubular injury and also inhibited the apoptosis of tubular epithelial cells in mice [ 99 ], showing potential clinical applications in settings where AKI might be anticipated, including kidney transplantation or cardiothoracic surgery. Furthermore, spermidine supplementation markedly attenuated kidney dysfunction and tubular damage after IRI by reducing oxidative stress, lipid peroxidation, and necrosis [ 100 ].…”