Computational protocol: Polyploidy Analysis and Attenuation of Oxidative Stress in Hepatic Tissue of STZ-Induced Diabetic Rats Treated with an Aqueous Extract of Vochysia rufa

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Protocol publication

[…] Adult Wistar male rats (200–250 g) were obtained from ANILAB (Animais de Laboratório Criação e Comércio Ltda-EPP), Paulínia, Brazil. The animals were maintained under standard conditions (22 ± 1°C, 60 ± 5% humidity and a 12 h light : 12 h dark cycle). The animals were fed a commercial pellet diet (65.82% carbohydrate, 5.36% fiber, 21.0% protein, and 4.96% fat) (BioBase, SC, Brazil) and received water ad libitum. All procedures for the handling, use, and euthanasia of the animals were approved by the Ethics Committee for Animal Research at the Federal University of Uberlandia, Brazil (CEUA/UFU 060/10). [...] Hepatocyte nuclei were isolated according to the method described by Blobel and Potter []. Immediately after tissue homogenization, using a Potter homogenizer in 0.25 M sucrose in TKM (Tris 0.02 M, KCl 0.025 M, and MgCl2 0.005 M) buffer, the homogenate was filtered, mixed with 2.3 M sucrose in TKM, and applied to 2.3 M sucrose in TKM solution. This process produced a gradient that was centrifuged at 124,000 ×g at 4°C for 30 min. Pelleted nuclei were suspended in TKM buffer and stained with propidium iodide for analysis by flow cytometry (BD Accuri C6 Flow Cytometer). Analysis was then performed using FlowJo software. […]

Pipeline specifications

Software tools Biobase, FlowJo
Applications Miscellaneous, Flow cytometry
Organisms Rattus norvegicus
Diseases Diabetes Mellitus, Hyperglycemia
Chemicals Carbohydrates, Glyburide, Streptozocin