Comparison of effifciency of treatment methods in valproic acid induced rat model of autism

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by regression in social skills, learning skills, language skills and successive pointless repetition. Although the prevalence of autism varies by countries and states (0.5-5 %), there is a rapid and dramatic increase in the incidence worldwide. VPA induced animal model of autism in rats was carried out in this study. Determination of whether autistic behaviour disorders exist or not, neuro-immunohistopathologic changes occured in the brain, IL1β, IL6, TNFα, GPX, MDA, and CASP3 levels in the brain tissue, relationship between autism and CC2D1A, CASP3 gene expression levels (RT-PCR) in the Control Group, and compare the obtained data statistically with Control Group were aimed in the study. In additon to these, investigation of whether fingolimod, suramin, and N-acetyl sistein used in the modern medicine treatments, and curcumin, resveratrol used in the complementary and alternative medicine treatments, have ameliorating effect on the autistic behaviour disorders or not, effects on the expression levels of CC2D1A and CASP3 genes, and comparison of results obtained from both the two treatment groups, with each other also were aimed in the study. Of the totally 30 female rats used in the study, while 25 were induced by VPA for autism modelling, 5 were not induced by VPA. Fourty eight pups delivered from the 25 mother rats were included to disease group, and eight pups delivered from 5 healty mother rats were included to the healthy control group. All the pups included to the disease group (48) were devided in to six subgroups that; five of them were consisted treatment groups and the one was disease (treatment) control group. One of the five agents were administered to each of the five treatment groups. No agent was administered to the disease (treatment) control group. Data obtained from the patient and treatment groups were statistically compared with control groups of each ones. Water Maze behaviour test were performed and scored in order to asses cognitive functions. IL1β, IL6, TNFα, GPX, MDA, and CASP3 levels were assayed by ELISA method in the brain tissue materials obtained after sacrification procedure. Expression levels of CC2D1A and CASP3 genes were determined in the brain tissue by RT-PCR method. Histopathologic microscobic examinations were made with slides prepared from brain tissue materials and stained with hematoxylin-eosin (H&E). Behaviour test results of the disease group and control group were found statistically significanty different (p<0,05). Statistically significant difference were found when comparing the results of the amelorative effects of the fingolimod, suramin, curcumin, and resveratrol groups with each disease (treatment) fourth of the control groups (p<0,05). Suramin administration created negative effect on behaviour test with statistically significantly difference degree. No statisticallysignificant difference were found when comparing of the N-acetyl sistein treatment group's results with the disease (treatment) control group (p>0,05). Chromatrolysis, focal glial cell infiltration, areas of neuronophagy, neuronal vacuolation, neuro inflammation were found in the brain with histopathological examinations. Results of ameliorative effects of histopathological abnormal brain findings; fingolimod, suramin, and N-acetyl sistein treatment groups compared with each treatment control group; all the treatment groups were found statistically different. The strongest ameliorative effect for histopathological brain abnormalities were found in the fingolimod treatment group. When all the ELISA assay results evaluated comperatively obtained from treatment groups and treatment control groups; only GPX (p<0,05) and CASP3 (p<0,05) test results were found to be statisticallysignificant difference at fingolimod treatment group. It was reported that etiology of the autism was related with genetic, epigenetic, and enviromental factors. Of the genetic factors CC2D1A gene; the coiled-coil and C2 domain containing protein 1A (CC2D1A) gene was identified as a repressor of serotonin receptor 1A (HTR1A) expression. Unfortuantely meaningful results were not able to found for CASP3 gene expression levels. CC2D1A expression level results of the disease group and control group compared to each other; statistically significant difference was found (p<0,05). When compared repressor effect of the treatment groups for CC2D1A gene expression with treatment control group; while fingolimod (p<0,05), curcumin (p<0,05), and resveratrol (p<0,05) were found to be statistically significant difference; statistically significant difference was not found at suramin (p>0,05) and N-acetyl sistein (p>0,05) treatment groups. Of all the treatment groups fingolimod has the strongest repressor effect on the CC2D1A gene expression, and curcumin and resveratrol have the same effect respectively. Different neuro-immunopathologic changes occured in the autistic brain. Development of autistic behavior disorders and gene expressions may be triggered by these pathologic changes, can be concluded from the study. In additon, the following results were also obtained; 1. Fingolimod used for modern medicine therapies, curcumin and resveratrol used in the alternative and complementary therapies were found as repressors of the CC2D1 gene, and fingolimod was the strongest repressor effect, fingolimod and resveratrol was found as the most effective two agents for amelioriation of the autistic disorders. 2. Suramin administration effected behaviour disorders as negativeley. 3. Expression elvels of CC2D1A gen has ciritical value, and hipo or hiper expressions trigerring negative results with survive of autism.