Ketogenic diet and epilepsy: a nutritional approach focused on the morphofunctional modulation of the brain-intestine axis and its lipid, oxidative and inflammatory aspects

• Grant number: 20/03529-6
• Support Opportunities: Regular Research Grants
• Start date: February 01, 2021
• End date: July 31, 2023
• Field of knowledge: Health Sciences - Nutrition - Nutrition Biochemistry
• Principal Investigator: Nágila Raquel Teixeira Damasceno
• Grantee: Nágila Raquel Teixeira Damasceno
• Host Institution: Faculdade de Saúde Pública (FSP). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Carla Alessandra Scorza Bahi ; Christian Hoffmann ; Debora Levy ; Geni Rodrigues Sampaio ; Lionel Fernel Gamarra Contreras ; Sayuri Miyamoto

Abstract

Epilepsy is the most common neurological disease in children. Classically, pharmacological polytherapy is the most accepted treatment with the best clinical results, however, about 30% of patients are resistant to this treatment. Since the 20s of the last century, the ketogenic diet (CD), formed by about 90% of fats, has been presented with an effective treatment in the control of convulsive crises of drug resistant patients. Despite its effectiveness, CD has shown a negative impact on the cardiometabolic risk of patients, where dyslipidemia, inflammatory and oxidative processes are important adverse effects. Studies have shown that modulation of the fatty acid profile can reduce these adverse effects, without reducing clinical efficacy. Given the above, the objective of the present proposal is to evaluate, through an experimental model of epilepsy induction, the impact of CD containing different fatty acid profiles on the brain-intestine axis, focusing on lipid, oxidative and inflammatory markers. After 50 days of epilepsy induction (lithium-pilocarpine, 30/300, mg/mg) and 3 months of monitoring, 60 male Wistar P40 rats will be distributed in the following groups: positive control (GC-R [+] or G1) will receive a standard ration for rats and will be induced to the epileptic state by lithium-pilocarpine; negative control (GC-R [-] or G2) will receive a standard ration for rats, however it will not induce epileptic states; Classic DC (DCC-R [+] or G3) will receive the Classic DC containing 90% fat; DC PUFAS (DCP-R [+] or G4) will receive a modified DC in the fatty acid profile (DC PUFAS); Classic DC w3 (25 mg DHA/kg body weight; DCCW-R [+] or G5) will receive Classic DC supplemented with omega 3 and, DC PUFAS w3 (DCPW-R [+] or G6) will receive DC PUFAS supplemented with omega 3. During this period, feed intake, weight gain and ketone body generation will be assessed weekly. The number of seizures (Racine scale) and motor and gait activity (Actimeter LE 8825 and Catwalker infrared laser system) will also be assessed at weekly intervals using video monitoring. At baseline and T90 days the animals will be kept in metabolic cages to collect feces for further analysis of the intestinal microbiota (17S), lipid profile (LC-MS/MS) and histopathological analysis (H/E). The following biomarkers will be monitored from plasma: erythrocyte fatty acids (CG/MS), ketone bodies (B-hydroxybutyrate), lipid and lipoprotein profile (total cholesterol, LDL-c, HDL-c, triglycerides, Apo AI, Apo B ) and non-esterified fatty acids (NEFAS). The brain will be removed from the skullcap and after appropriate treatments, the following analyzes will be performed: fatty acid profile (CG/MS), lipid analysis (LC-MS/MS), expression and synthesis of neuroinflammatory cytokines (BDNF and MNDA, IL1b , IL6, IL8, IFN gamma), TNF-a and anti-inflammatory (IL10), and histopathological analysis (H/E) and oxidation markers (LDLox, ORAC, Lag time). Intestinal fragments will be resected for analysis of oxidative (LDLox, ORAC, Lag time), inflammatory and intestinal barrier integrity (LPS, FBAP and 1-beta lithostamine) markers. (AU)