An improved method for the establishment of an autistic mouse model with down-regulation of PTEN gene
10.19405/j.cnki.issn1000-1492.2025.03.011
- VernacularTitle:一种PTEN基因下调的孤独症幼鼠模型构建的改良方法
- Author:
Weili DANG
1
;
Lvyuan LIANG
1
;
Jialei CAO
1
;
Shenao DING
1
;
Bingqi WEI
1
;
Xia QIU
1
;
Bingxiang MA
1
Author Information
1. 河南中医药大学第一附属医院儿科医院,郑州 450099;河南中医药大学儿科医学院,郑州 450046
- Publication Type:Journal Article
- Keywords:
PTEN;
autism spectrum disorder;
animal model;
valproic acid
- From:
Acta Universitatis Medicinalis Anhui
2025;60(3):462-471
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a PTEN gene-downregulated ASD juvenile rat model(VPA-ADV)through combined application of valproic acid(VPA)and PTEN adenovirus(ADV),then to compare the newly constructed animal model with two traditional autistic animal models of VPA and AD,and ultimately to prove the advantages of this model in animal model establishment of acupuncture treatment for ASD.Methods Wista rats at 12.5 days of gestation were randomly divided into 2 groups.VPA rats were given 600 mg/kg of normal saline(NS)intraperito-neally.Weight,eye opening time and tail deformity were recorded.The newborn mice in NS group were randomly divided into three groups(10 rats in each group):normal(NS)group,virus(ADV)group and virus-negative control(ADV-NC)group;VPA group(20 young rats)was randomly divided into 2 groups(10 rats in each group):valproic acid(VPA)group and valproic acid-binding virus interference(VPA+ADV)group.The body weight,tail length,curved tail,geotaxis text time and skeletal deformity of 5 groups of young rats after birth,the neurobehavioral behavior of 21-day-old rats,and the myelin structure of brain tissue under electron microscope were observed,the expression levels of PTEN,PI3K,AKT,GSK3β and MBP were detected by immunohistochemistry,Western blot and q-PCR.Results Compared with the NS group,the VPA group had significantly increased mal-formation rate,slow weight gain,slow tail length growth,and long negative geotaxis reflex time(P＜0.05).Com-pared with the NS group,the weight gain,tail length growth and negative geotaxis reflex time of the three model groups were slower(P＜0.05),and the performance of VPA+ADV model was the most significant.There were significant differences in the time of crossing the central grid,the number of crossing the edge grid,and the time of crossing the edge grid in the open field test between the three groups and the NS group(P＜0.05).In the self-grooming experiment,the number of interactions in VPA-ADV model was the least(P＜0.05),and the number of digging and self-grooming was the most(P＜0.05).In the three-box social experiment,VPA-ADV model had the shortest average number of entries into the social box and the shortest residence time(P＜0.05),the time of find-ing the platform in the water maze experiment was the longest,and the number of times crossing the third quadrant was the least(P＜0.05).The structure of the myelin sheath layer in the corpus callosum was observed by transmis-sion electron microscopy.The structure of the NS group was clear and complete.Compared with the NS group,the myelin structure of the ADV-NC group was similar,and the myelin structure of the three model groups was stratified and broken,and there were pathological changes and myelin damage in the ASD.Among them,the myelin sheath of the VPA-ADV model was thickened,stratified,and severe visible disintegration.The results of immunohisto-chemistry,Western blot and q-PCR showed that the expression of PTEN in VPA+ADV model was down-regulated by about 50％,which was the most obvious.The expression of AKT and MBP increased,and the expression of GSK3β decreased(P＜0.05).However,the results of q-PCR showed that the expression of PI3K-mRNA in the three model groups significantly increased(P＜0.05),and the change of VPA+ADV model was the most signifi-cant.Conclusion The novel PTEN-ASD animal model established by VPA+ADV method is observed to have sig-nificant pathological changes that are typical of the manifestation of ASD myelin dysplasia and is determined to have better results than the two traditional autistic animal models.
