Elsevier

Biomedicine & Pharmacotherapy

Volume 83, October 2016, Pages 392-396
Biomedicine & Pharmacotherapy

The efficacy of Aesculus hippocastanum seeds on diabetic nephropathy in a streptozotocin-induced diabetic rat model

https://doi.org/10.1016/j.biopha.2016.06.055Get rights and content

Abstract

Cytokines, such as transforming growth factor (TGF)-ß1, and increased oxidative stress are considered to be responsible for the development of diabetic nephropathy. We hypothesized that Aesculus hippocastanum (AH) seeds may have preventive effects on oxidative stress and TGF-β-related diabetic nephropathy in streptozotocin (STZ)-induced diabetic nephropathy in rats. Twenty-one male Sprague-Dawley albino rats were divided into three groups (n = 7). Except for the control group, they all had diabetic nephropathy induced by an intraperitoneal injection of STZ. While the diabetes group did not receive any medication, the diabetes + AH group was given the medication for 4 weeks. After the experiment, analyses were performed to evaluate the glomerular area, severity of sclerosis, and fibronectin immunoexpression, as well as levels of malondialdehyde (MDA), TGF-β, blood urea nitrogen (BUN), blood glucose, creatinine, and proteinuria. It was found that glomerular area, severity of sclerosis, fibronectin immunoexpression, and levels of MDA, TGF-β, BUN, creatinine, and proteinuria were decreased in the diabetes + AH group. It is known that diabetic nephropathy is induced, to a large extent, by hyperglycemia. In the present study, AH extract ameliorated diabetic nephropathy without decrease in blood glucose levels. In the study, AH seeds showed beneficial effects on the functional properties of the kidney and microscopic improvements in diabetic nephropathy.

Introduction

Diabetic nephropathy (DN) is a common, severe complication of diabetes, developing in 20–40% of diabetic patients [1]. DN is characterized by diffuse glomerulosclerosis and nephrotic syndrome, which are caused by microangiopathy of capillaries in the kidney glomeruli [2].

There are several mechanisms suggested for the pathophysiology of DN. Glomerular hyperfiltration and hyperperfusion represent one mechanism related with high blood glucose levels [3]. Glomerular glucose hyperfiltration causes NaCl loss and results in decreased NaCl presentation to the macula densa. Activated macula densa–mediated feedback mechanisms increase the levels of angiotensin II, which in turn causes glomerular fibrosis and sclerosis with the stimulation of transforming growth factor (TGF)-ß1 (a hypertrophic, prosclerotic cytokine) [4], [5]. Another postulated mechanism for DN is the nonenzymatic glycosylation of tissue proteins resulting from chronic hyperglycemia. The glomerular basal membrane is adversely affected by the formation and deposition of advanced glycation end-products [6]. In addition, these advanced products can alter signal transduction, which can be involved in the pathogenesis of DN via alteration of signaling molecules, such as cytokines, hormones, and free radicals [1].

Another postulated mechanism is chronic hyperglycemia–induced protein kinase C (PKC) activation. This has been associated with alterations in blood flow in renal and glomerular vessels, extracellular matrix expansion, basement membrane thickening, hyperfiltration, enhanced angiogenesis, and excessive apoptosis [7]. PKC activation leads to increased activity of mitogen-activated protein kinase (MAPK) [8]. MAPK activity, PKC activity, and TGF-β1 activation, which is commonly related with oxidative stress and reactive oxygen species (ROS), cause hyperfiltration and glomerular basement membrane thickening [9].

Recent studies have tried to define the pathogenesis of DN via oxidative stress mechanisms. Even before symptoms of diabetes become clinically apparent, hyperglycemia specifically induces oxidative stress [10]. The first event that results in kidney damage is the increased production of reactive oxygen species into the mitochondria due to the oversupply of NADH and FADH2 in the oxidative phosphorylation chain [11]. This is the result of an oversupply of the Krebs’ cycle with pyruvate, which is in turn the result of an increased glucose influx into the cell. The above sequence of events takes place in renal endothelial cells, which are unable to downregulate the glucose transporter-1 despite the hyperglycemic environment [12]. From this event originates a cascade of other detrimental events, such as PKC activation, production of advanced glycation end-products, TGF-β upregulation, and cytosolic NAPDPH consumption. In previous studies, products of lipoxidation in the mesangial matrix was detected via histological analysis of human kidney biopsy specimens [13], [14].

The seeds of the Aesculus hippocastanum (AH) plant (horse chestnut) are used as a drug to treat venous disorders, such as varicose veins, phlebitis, leg ulcers, and hemorrhoids. AH is an astringent and anti-inflammatory plant that helps to decrease the symptoms of certain illnesses, such as arthritis, strains, and tendonitis [15], [16]. Recent studies have shown that AH seeds can decrease the oxidative stress and anti-inflammation effects resulting from decreased levels of cytokines like tumor necrosis factor (TNF)-α and interleukin (IL)-1β in different tissues [17], [18], [19].

In this study, we hypothesized that AH seeds may have preventive effects on oxidative stress and TGF-β-related DN in streptozotocin (STZ)-induced DN in rats.

Section snippets

Animals

Approval for all procedures in the experiment was received from the Ege University Animal Research Ethics Committee. In the study, we used 21 male Sprague Dawley rats. The rats weighed 200–220 g. The animals were fed with standard rat food pellets (Abalim®, Abalioglu Food Inc., Denizli, Turkey) ad libitum and kept in cages with standard laboratory conditions. Animals with blood glucose levels higher than 120 mg/ml were not included in the experiment.

Suspension of A. hippocastanum seeds

In the study, Venotrex® tablets (Abdi Ibrahim,

Results

Histopathological and immunohistochemical examinations of renal tissues are given in Fig. 1. The results of the analysis of the glomerular area (% of control), severity of sclerosis, fibronectin immunoexpression (percent), and levels of MDA, TGF-β, proteinuria, BUN, creatinine, and blood glucose are given in Table 1.

In histopathological and immunohistochemical examinations of renal tissues, a thickening of the glomerular basement membrane and an increase in the mesangial matrix were seen in the

Discussion

In the study, we attempted to prevent STZ-induced DN through the administration of AH seeds in rats. The results showed that AH seeds could prevent the development of STZ-induced DN in rats.

In the literature, when AH seeds were systemically administered to experimental animals, antioxidant effects were shown in different tissues like the kidneys, liver, ileum [17], [18], [19], [28], [29]. In our study, we showed the powerful antioxidant effects of AH seeds on MDA measurements. In addition, AH

Conflict of interest

None of the authors has a conflict of interest to declare.

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