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Protective Effect of Vaccinium Arctostaphylos L. Fruit Extract on Gentamicin-Induced Nephrotoxicity in Rats

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Protective Effect of Vaccinium Arctostaphylos L. Fruit Extract on Gentamicin-Induced Nephrotoxicity in Rats Downloaded from http://iranpaper.ir http://www.itrans24.com/landing1.html Comparative Clinical Pathology https://doi.org/10.1007/s00580-018-2743-0 ORIGINAL ARTICLE Protective effect of Vaccinium arctostaphylos L. fruit extract on gentamicin-induced nephrotoxicity in rats Mohsen Akbari Bazm1 & Mozafar Khazaei2 & Elham Ghanbari2 & Leila Naseri1 Received: 22 March 2018 /Accepted: 16 May 2018 # Springer-Verlag London Ltd., part of Springer Nature 2018 Abstract Aminoglycosides are extensively used owing to advantages such as low cost and availability. Nephrotoxicity induced by antibiotics has been analyzed in different animal models. This study was aimed to investigate the effect of Vaccinium arctostaphylos L. fruit extract on gentamicin-induced nephrotoxicity in rats. In this experimental study, 25 rats were divided into five groups, including negative control (0.5 ml normal saline), positive control (80 mg/kg gentamicin and 0.5 ml normal saline), and three Vaccinium arctostaphylos L. fruit extract groups at 100, 200, and 400 mg/kg concentrations plus 80 mg/kg gentamicin. All administrations were done intraperitoneally for 7 days. Thirty days after the first injection, blood samples were taken from the heart. Serum creatinine (Cr), blood urea nitrogen (BUN), and sodium and potassium levels were measured. To prepare histological sections, the kidneys were kept in 10% formalin. Having prepared the histological sections, the volume of renal tubules (proximal and distal convoluted tubules and loop of Henle), renal vessels, interstitial tissue, and number and volume of renal glomeruli were measured. Vaccinium arctostaphylos L. fruit extract significantly increased serum urea (p =0.023), creatinine (p = 0.009), volume of proximal convoluted tubules (p = 0.02), volume of distal convoluted tubules (p = 0.032), loop of Henle (p = 0.038), and glomeruli (p = 0.015). However, it significantly reduced the volume of vessels (p = 0.009) and inter- stitial tissue (p = 0.001). The 400 mg/kg concentration exerted the highest effect. Vaccinium arctostaphylos L. fruit extract has protective effects against gentamicin-induced renal impairments. Keywords Kidney . Vaccinium arctostaphylos L. Gentamicin . Stereology Introduction erythropoietin, which is effective in stimulating the production of erythrocytes, and gluconeogenesis are performed with co- Kidneys play a key role in establishing physiological balance, operation of the kidneys. Acute injury to the kidney leads to regulating the balance of water and electrolytes, excreting changes in the ratio of body ions, which in turn results in metabolic and bioactive waste like many drugs, and regulating blood acid-base imbalance (Coca et al. 2008). arterial blood pressure by secreting renin. Secretion of Various factors such as aminoglycoside antibiotics cause acute renal impairments. Gentamicin is an antibiotic from this family that leads to acute renal impairments despite effects on * Leila Naseri infections induced by gram-positive and -negative bacteria [email protected] (Rodieux et al. 2015). The destructive effect of gentamicin Mohsen Akbari Bazm can be observed through measurement of serum urea and cre- [email protected] atinine levels of patients. Gentamicin has been used in many Mozafar Khazaei studies as one of the toxic compounds inducing acute renal [email protected] dysfunctions (Morin et al. 1980). Production of free radicals Elham Ghanbari and nitrogen compounds by these antibiotics causes renal tox- [email protected] icity. Reactive oxygen species (ROS) causes lipid peroxida- tion of cell membrane, denaturation of proteins, DNA damage 1 Department of Anatomical Sciences, Medical School, Kermanshah and apoptotic induction, and cell death. Gentamicin is able to University of Medical Sciences, Kermanshah, Iran induce histological damage by reducing the activity of antiox- 2 Fertility and Infertility Research Center, Kermanshah University of idant enzymes such as catalase (CAT), superoxide dismutase Medical Sciences, Kermanshah, Iran Downloaded from http://iranpaper.ir http://www.itrans24.com/landing1.html Comp Clin Pathol (SOD), and glutathione peroxidase (GPX) in the kidneys (El- approved by a botanist from Razi University, Kermanshah. Razek and Kamel 2011). Antioxidant compounds decrease The fruit was dried and powdered by a mill, and 1500 g of the release of free radicals and inhibit histological damage. powder was kept in 70% ethyl alcohol solution for 72 h. The Various plant extracts and compounds with antioxidant prop- solution was then filtered by filter paper (Whatman U.K. 2). A erties have been used to improve toxicity due to toxic com- vacuum distillation apparatus (Heidolph Collegiate, pounds such as gentamicin, cisplatin, and acetaminophen LABOROTA 4000, Germany) was sued to condensate the (Raoofietal.2015; Khoursandi and Ourazizadeh 2016; solution at 50 °C. The obtained extract was kept at 4 °C Khazaei et al. 2012). (Khazaei et al. 2006). A total of 25 male Wistar rats with the Vaccinium arctostaphylos L., a member of Ericaceae fam- mean weight of 130 ± 10 g were used in this study. The ani- ily, is a shrub with no thorns and with heart-shaped leaves and mals were kept at 12/12 dark/light cycle at 22–35 °C and toothed edges found in the northern and northwestern parts of humidity of 40–65, with free access to standard pellet. The Iran. Vaccinium arctostaphylos L. is native to Iran, and other rats were kept for 3 days to adapt to the environment. species of this plant (Vaccinium myrtillus)arefoundin The study groups consisted of five groups, each with five Georgia and Russia. This plant is inflorescence and contains rats, including negative control receiving 0.5 ml normal sa- 20–40 flowers. The plant fruit constitutes the pharmaceutical line, positive control receiving 80 mg/kg gentamicin, and part of it, which is in red to dark purple and matures in late three case groups receiving 80 mg/kg gentamicin plus summer or early fall (Akhondzadeh 2000). Vaccinium arctostaphylos L. extract at 100, 200, and Studies have documented the presence of anthocyanin, 400 mg/kg concentrations (Feshani et al. 2011; Baradaran et phenolic, flavonoid, and coumarin compounds in the fruit al. 2014;Ismailetal.1994). All administrations were done and leaves of this plant. Also, glucose compounds and organic intraperitoneally. At the end of the study (30 days after the first acids constitute the components of the fruit of this plant administration), the animals were anesthetized, and blood (Nickavar et al. 2002). These pigments and generally phenolic samples were taken from the heart. Serum urea (BUN) and compounds present in the cells of this plant protect it from creatinine levels were measured by Pars Azmoon assays and ultraviolet ray and insects’ attacks. spectrophotometry method (SINCO), and serum sodium and In traditional medicine, this plant has been used to reduce potassium ions were measured by photometric method blood sugar and lipid and to treat hypertension (Hakimi et al. (Jenway Flame photometer PFP) (Hald 1947). Then, the kid- 2014;Su2012). The most important antioxidant compound of neys were extracted from the body and weighed by a digital this plant is anthocyanin. This compound inhibits the free scale (AND FA 2014, China). radicals and apoptosis and prevents the oxidation of cell mem- The samples were preserved in 10% formalin for 72 h. The brane lipids (Akhondzadeh 2000). volume of the kidneys was determined by Cavallier’s method Researchers have reported antioxidant and anticancer prop- after estimating the tissue shrinkage rate. To this end, first, the erties for anthocyanins in reducing coronary diseases initial volume of the kidneys was measured by immersion (Nickavar and Amin 2004). Flavonoids are antimicrobial, an- technique, and isotropic uniform random (IUR) sections were tiviral, anti-atherosclerotic, antidiabetic, antioxidant, and anti- prepared by orientator method. In this method, the samples inflammatory compounds that exert their effects through inhi- were placed on circles divided into ten equal parts, and each bition of enzymes responsible for production of superoxide sample in the first section was cut into two halves by selecting anions such as xanthine oxidase and protein kinase C a random number from 0 to 10. Then, the surface of each (Nickavar and Amin 2004; Middleton and Kandaswami section was placed parallel to another circle with previous 2000; Robak and Gryglewski 1988). Vaccinium specifications; and the same as the former stage, another sec- arctostaphylos L. fruit is proposed as a medicinal plant owing tion was prepared, and 2-mm sections were prepared from the to important antioxidant compounds. This study was conduct- sections of previous stage. Next, 6–8sectionswererandomly ed to investigate the protective effect of Vaccinium selected from each kidney (Gundersen and Jensen 1987; arctostaphylos L. fruit extract on serum and morphologic fac- Akbari et al. 2017). A circle was prepared from one of the tors of the kidney following gentamicin-induced renal sections by punching, and its diameter was measured by cal- impairment. iper. After that, each section was subjected to conventional successive stages of tissue passage and was molded in molten paraffin. Then, 5-μ sections were prepared from it by a rotary Materials and methods microtome (Leica RM2235, USA). To measure tissue shrink- age, the final diameter of circular section was measured after Plant tissue passage stages and was inserted in the following rela- tion (Gundersen et al. 1988):
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