Effects Of Ginsenoside RG1 On Behavior And NOX2/ROS Pathway In Autism Model Rats

Objective: To investigate the effects of Ginsenoside RG1 intervention on abnormal behavior and NOX2/ROS pathway in autism model rats.Methods: Clean Wistar rats were selected,including 10 males and 20 females.According to the preparation method of Schneider autism model rats,the female rats were intraperitoneally injected 600 mg/kg sodium Valproate(VPA)at 12.5 days of gestation.The pregnant rats in the normal group were intraperitoneally injected with the same amount of normal saline at the same time.The developmental and behavioral tests of the offspring of the model group were carried out to identify whether the model was successfully prepared.Developmental detection methods:body weight,brain weight,directional tendency,eye opening time;Behavioral testing methods include three box test,social communication test and repeated stereotyped behavior test.50 mice in the normal group,the model group and the drug group were randomly selected.The drug group was intraperitoneally injected with ginsenoside RG1(40 mg/kg,IP)from day 7 for 3weeks,and the normal group and model group were injected with the same dose of normal saline.Morris Water Maze test learning and memory ability,other behavioral development test methods are the same as above;HE staining was used to observe the pathological changes in CA1,CA3 and DG areas of the hippocampus for morphological detection.The number of reactiveoxygen species(ROS)was detected by immunofluorescence staining.MDA expression level was detected by kit.Western blot was used to detect the expression levels of NOX2 and pP47 phox,components of NOX2 oxidase.Results: 1.Identification of mouse models of autism: the body weight and brain weight of the mice in the autism model were significantly lower than those in the NS group at different time points,and the time to open eyes was delayed,and vestibular and motor development was backward(P<0.05).In this study,three box experiment,social communication experiment and repetitive stereotypical behavior experiment were used to detect the core symptoms of ASD in model rats on P28.The results showed that the cumulative time of repetitive behavior increased,the olfactory resolution decreased,the preference for social odor decreased,and the social interaction ability and social novelty ability were impaired(P<0.05).2.Effects of ginsenoside RG1 on behavioral development in model group: The results showed that compared with VPA group,the body weight of offspring rats in Rg1 group was significantly heavier than that in VPA group at P28,the accumulative time of stereotypic action was reduced,the time of smelling social odor was increased,and the learning and memory ability was significantly improved(P<0.05).3.Effects of ginsenoside RG1 on morphology and oxidative stress in hippocampus of model rats: HE staining showed that nerve cells in CA1,CA3 and DG areas of hippocampal tissue in model group were arranged loosely and disordered,and cells were swollen.After ginsenoside RG1 treatment,the pathological changes were relieved.Immunofluorescence and kit results showed that: At P7 and P28 days of age,the production of ROS and MDA was higher in VPA group(P<0.05),and the levels of ROS and MDA in hippocampus were down-regulated after ginsenoside Rg1 intervention(P<0.05).Western blot analysis showed that the protein expression levels of NOX2 and p-P47 phox in hippocampus of VPA group were higher than those of normal group at P7 and P28 days of age(P<0.05).Compared with VPA group,the expression levels of NOX2 and p-P47 phox in the hippocampus of Rg1 group were down-regulated,but the protein expression of NOX2 was not significantly decreased(P>0.05),while the protein expression of p-P47 phox was significantly decreased(P<0.05).Conclusion: 1.The production of ROS and MDA and the expression of NOX2 and pP47 phox protein in hippocampus of autism model mice increased;2.Ginsenoside RG1 can significantly reduce the production of ROS and MDA in hippocampus of autism model mice,inhibit the expression of p-P47 phox protein,and play a neuroprotective role;3.Ginsenoside RG1 can improve the learning and memory abilities of autism model mice,which may be related to the inhibition of NOX2/ROS pathway and the reduction of ROS production.