Protective Effects Of Resveratrol Against Oxidative Stress Damage In Gcdh^-/- Rats

Background and objectiveGlutaric acidemia typeⅠis an autosomal recessive genetic neural metabolic disorders caused by a mutation in a gene encoding Glutaryl-CoA Dehydrogenase（GCDH）.The enzymatic defect results in metabolism disorder of lysine,hydroxyl lysine and tryptophan,and elevated concentrations of glutaric acid,3-hydroxyglutaric acid and glutaric acid pentene diacid in the body fluids and tissues,accumulated nervous system damage in the brain.Imbalance of reactive oxygen species（ros）leads to oxidative damage of cells and tissues,and oxidative stress injury is closely related to a variety of neurodegenerative diseases.Oxidative stress impairs cell membrane potential,initiates pro-inflammatory and pro-apoptotic signals,and leads to a vicious cycle of oxidative stress and neuroinflammation.Resveratrol（RESV）is a natural non-flavonoid polyphenolic compound widely found in plants such as Vitis,Brassica,Peanut,and Cucurbita.RESV has powerful anti-oxidant and anti-inflammatory properties,protecting the heart,protecting nerves,chemotherapy and anti-aging.RESV is known as a neuroprotective agent that protects against oxidative damage in neurons and glial cells.In the experiment,a successful GCDH gene knockout rat model was constructed,and the genotype of the mice was identified after mating,and the mice were fed a normal diet or a Lysine（Lys）diet.The RESV protectant was given to observe the changes in body weight,pathological changes in cerebral cortex,hippocampus and striatum,and oxidative stress indexes of ROS,MDA and GPx were detected.The incidence of Gcdh^-/-rats,the changes of oxidative indexes in vivo and the anti-oxidation and neuroprotective effects of RESV on Gcdh^-/-rats were observed from animal model level,which provided experimental basis for clinical intervention and treatment of GAI.Methods1.Gcdh^-/-,heterozygous and wild type rat were bred and multiplied,and genotypes were identified by PCR at 3 weeks of age.2.The genotypes of rat were randomly divided into 6 groups,normal diet group of wild rats（N）,high Lys group of wild rats（N+Lys）,high Lys+RESV group of wild rats（N+Lys+RESV）,Gcdh^-/-normal diet group(Gcdh^-/-),Gcdh^-/-high Lys Group(Gcdh^-/-+Lys),Gcdh^-/-high Lys+RESV group(Gcdh^-/-+Lys+RESV).3.At the age of 4 weeks,rats in the Lys group and the Lys+RESV group were given a high lysine diet（4.7%high lysine）.At 10:00 am,the rats in the high Lys+RESV group were given a 100mg/kg resveratrol gavage,and the rest rats were given normal saline gavage.Specimens were collected after 4 weeks of continuous administration.4.The deaths of the rats in each group were observed.The daily weight gain of the rats in each group was statistically analyzed.The morphological changes of the frontal cortex and hippocampus were observed by HE staining.The changes of neurons and astrocytes were observed by immunofluorescence staining GFAP,NeuN and S100β.The oxidative stress indicators of frontotemporal cortex and hippocampus（SOD,MDA and GPx）were detected.Results1.The genotypes of the progeny rats after breeding were divided into wild type,heterozygous and homozygous.2.The survival rate and body weight growth rate of Gcdh^-/-high Lys group and Gcdh^-/-high Lys+RESV group were significantly lower than those of wild type and homozygous normal diet group.The survival rate of Gcdh^-/-high Lys+RESV group was higher than that of Gcdh^-/-high Lys group,and there was no significant difference in body weight.3.HE showed that the Gcdh^-/-rats in the high Lys group had more disordered cell arrangement and more vacuolization than the other five groups,and immunofluorescence staining showed that the GFAP and S100βin the frontotemporal cortex of the high Lys group were more than those in the normal diet group and NeuN was less than that in the normal diet group.The morphology of Gcdh^-/-high Lys+RESV group was significantly improved compared with Gcdh^-/-high Lys group,NeuN increased,and GFAP and S100βdecreased.4.In the Gcdh^-/-high Lys group,MDA in the frontotemporal cortex and hippocampus was significantly higher than Gcdh^-/-high Lys+RESV,GPx activity was decreased,P<0.05,SOD activity was no statistical difference.There was no statistical difference between the Gcdh^-/-high Lys+RESV group and the normal diet group.Conclusion1.Gcdh^-/-rats given a high Lys diet cause oxidative stress,leading to nervous system damage.2.RESV has anti-oxidative and neuroprotective effects on Gcdh^-/-rats.