Increase of Oxidative Stress and Accumulation of α-Synuclein in Wistar Rat’s Midbrain Treated with Rotenone

Abstract

Abstract.A neurodegenerative accumulation of α-synuclein effect of rotenone on the increase synuclein in the midbrain of Wistar rats. Thirty 250 g) were divided into three groups: the Blank group, the Solvent group (intraperitoneally injected with 1 ml/kg body weight of sunflower seed oil), and the Rotenone group (intraperitoneally injected with 2.5 mg/kg body weight of rotenone) for 9, 19, and 28 days. The rats were for the 9-day treatment, day 20 for the 19 for the 28-day treatment (2 rats/gr midbrains were isolated and extracted. Glutathione assay and α tests were performed. The results showed that the average oxidative stress index was highest in the Blank group (0.95 ± 0.24, 0.63 ± respectively). Meanwhile, groups (0.09 ± 0.03, 0.15 ± 0.03, and 0.13 ± 0.02 ng/mg tissue, respectively). Correlation analysis showed that the oxidative stress index proportional to the concentration of α midbrain of Wistar rats treated with rotenone indicated oxidative stress and led to an accumulation of α-synuclein protein. Here, we report our findings on the on the increase of oxidative stress and accumulation of α-synuclein in the midbrain of Wistar rats. Thirty six male rats (8-9 weeks, 200-were divided into three groups: the Blank group, the Solvent-rotenone group (intraperitoneally injected with 1 ml/kg body weight of sunflower seed tenone group (intraperitoneally injected with 2.5 mg/kg body weight of rotenone) for 9, 19, and 28 days. The rats were decapitated on day 10 day treatment, day 20 for the 19-day treatment, day 30, 40, 50 and 60 day treatment (2 rats/group or 6 rats/day of observation time). The midbrains were isolated and extracted. Glutathione assay and α-synuclein ELISA tests were performed. The results showed that the average oxidative stress index was highest in the Blank group (0.95 ± 0.24, 0.63 ± 0.23, 0.81 ± 0.27, Meanwhile, the concentration of α-Synuclein had decreased in all (0.09 ± 0.03, 0.15 ± 0.03, and 0.13 ± 0.02 ng/mg tissue, respectively). Correlation analysis showed that the oxidative stress index was inversely ortional to the concentration of α-synuclein. Our conclusion is that the midbrain of Wistar rats treated with rotenone indicated oxidative stress and led synuclein protein. Keyword

Protofibril formation: decreased total glutathione concentration as an early indicator of neuron damage in the brainstems of Wistar rats treated with rotenone

“…Breeding, maintenance, treatment, and sacrifice protocols were adapted to the provisions of the Health Research Ethics Committee, Faculty of Medicine, UNPAD-RSUP Dr. Hasan Sadikin Central General Hospital, Bandung, Indonesia (214/FKUP-RSHS/KEPK/Kep./EC/2010). 8 Materials Rotenone (R8875), sunflower seed oil (S5007), 5,5 0 -dithiobis (2-nitrobenzoic acid) (DTNB, D8130), reduced L-glutathione (G4251), glutathione reductase (G3664), NADPH (N5130) and thioflavin S (T1892) were purchased from Sigma-Aldrich or Merck.…”

“…The selected rats were divided equally into two studies-determination of total glutathione and protofibril examination. Four independent variables were examined: 1) rat treatment group: blank (not treated), solvent-rotenone (intraperitoneally administered sunflower seed oil at a dose of 1 mL/kg body weight/day), 8,21 and rotenone (intraperitoneally administered rotenone at a dose of 2.5 mg/kg body weight/day) groups 18 ; 2) duration of rotenone treatment to examine subacute and subchronic exposure (9, 19 and 28 days); 3) observation time (days 10, 20, 30, 40, 50, and 60); and 4) the location in the brainstem (upper and lower brainstem). 22 The concentrations of total glutathione and protofibrils were dependent variables (https://figshare.com/articles/dataset/Arrive-ten-aby/16880536).…”

“…DTNB was added and the absorbance of the solution was measured at λ = 412 nm (the by-product formed from the reaction of GSH and DTNB is yellow in solution) using a UV-vis spectrophotometer. 8 Brainstem preparation Thirty-six male rats were sacrificed according to the cardiac perfusion method using ketamine (90mg/kg body weight) and xylazine (10 mg/kg body weight) administered by intraperitoneal injection (27-gauge needle and 1-mL syringe). A cardiac perfusion with ice-cold PBS was performed followed by perfusion of a 4% paraformaldehyde solution containing 0.1% glutaraldehyde.…”

“…The sum of reduced GSH and oxidized GSSG is referred to as "total glutathione". [5][6][7][8] The concentration of total glutathione describes the capacity of the redox state in the cell. An increase in concentration of total glutathione indicates an increased ability of a cell to scavenge free radicals and protect the redox system, thereby protecting the cell from exposure to toxins.…”

“…is lipophilic and readily penetrates cell membranes, 19,20 causing neurons to undergo oxidative stress. 8 The aims of this research were to determine the total glutathione concentration in the upper and lower brainstems of Wistar rats treated with rotenone, and whether the sub-chronic exposure of rotenone causes the formation of protofibrils that, in humans, leads to the formation of Lewy bodies, as occurs in Parkinson's disease. The results reinforce the concept of pathogenesis of sporadic Parkinson's disease in rats treated with rotenone.…”