Investigating the effects of Cinnamomum-cassia nanoparticle conjugate on the Histomorphology of the kidney in type 2 diabetic rats.

Abstract

Introduction Diabetic nephropathy remains one of the biggest complications of diabetes. The incidence is increasing and more patients are experiencing progressive kidney failure due to lack of hyperefficient treatment. This study investigated the antidiabetic activity of Cinnamomum cassia silver nanoparticles (AgNPs) [(CcAgNPs)] and its effects on the kidneys of Sprague-Dawley rats induced with type 2 diabetes following administration of Streptozotozin. Materials and methods Adult healthy, pathogen-free male Sprague-Dawley rats, of a total number of 65 (N=65), weighing 250.0 ± 20 g were divided into 10 groups. Groups A-E (positive controls) consists of 30 rats, with 6 rats per group and the experimental groups F-J, consists of 35 rats, with 7 animals per group. Diabetes was induced in animals using Streptozotocin 60 mg/kg administered intraperitoneally. The animals were subjected to various treatments with Cc (100 mg/kg and 200 mg/kg) and CcAgNPs (5 mg/kg and 10 mg/kg). The treatments were administered orally using orogastric gavage and administration was carried out daily following treatment protocol for 56 days. The selected protocol for the experiment was officially approved by the Animal Ethics Committee (protocol reference number: AREC/74/016D). Cinnamomum cassia Silver Nanoparticles (CcAgNPs) was synthesized using the green option and characterized using UV (ultraviolet)–TEM (Transmission electron microscopy)-FTIR (Fourier-transform infrared spectroscopy) –XRD (X-ray powder diffraction), prior to administration. The animals were sacrificed on day 56. Blood and urine samples were collected for biochemical analysis. The kidneys were examined for histopathological changes using Hematoxylin and Eosin (H&E), periodic acid Schiff and Masson’s trichrome staining. Transmission Electron Microscope (TEM) and Stereological studies were carried out as well. Results Urinalysis showed extensive protein and albumin deposits in the urine. Ketones and nitrites levels which are markers of renal function were significantly lower (p< 0.05) in groups treated with CcAgNPs compared to negative controls. Urea and creatinine were also significantly (p < 0.05) reduced in treated groups compared to negative controls. The levels of reduced glutathione (GSH) was significantly different across all groups (p < 0.05). Serum Malondialdehyde (MDA) concentrations were significantly (p < 0.05) lower in CcAgNPs compared to controls. Liver enzymes (alanine aminotransferase) ALT was reduced significantly in groups treated with a low dose of CcAgNPs compared to negative controls. In the group treated with high dose (10 mg/kg) of CcAgNPs, (Aspartate transaminase) AST levels were significantly lower (p < 0.05), compared to the group treated with Cc (Cinnamomum cassia) and to the negative control. Stereological studies showed significantly decreased (p < 0.05) number of glomeruli and tubules in groups treated with Cc and CcAgNPs, compared to the negative control. Transmission Electron Microscope (TEM) revealed the thickness of glomerular basement membrane, in experimental groups, compared to positive controls. Histopathology of renal tissue showed severe glomerular distortion, tubular lesions with H & E and thickening of the basement membrane; pyknotic nuclei and vacuolization with PAS and MT, in the untreated negative control group. Positive controls showed regular glomeruli with normal Bowman’s capsular space, normal basement membrane and regular capillary network compared to negative controls. The degree of histopathological changes in the glomeruli and tubules appear to be dose-dependent.

Conclusion Diabetes negatively alters the cytoarchitecture and biochemistry of the kidneys of Sprague-Dawley rats while Cinnamomum cassia Silver Nanoparticles have the potential to ameliorate these changes. The possible pathway involved CcAgNPs may provoke the release of insulin-like, as well as the thioredoxin (Trx), which is one of the central antioxidants that can alleviate renal injuries in diabetic nephropathy. Keywords: Cinnamomum cassia; silver nanoparticles; diabetes; histomorphology