Regulation effect of Guipi Decoction on glutamate and PI3K/Akt/mTOR signaling pathway in rats with heart-spleen deficiency autism spectrum disorder
Objective It is to observe the effects of Guipi Decoction on the expression of related proteins of phosphati-dylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway in rats with heart-spleen deficiency autism spectrum disorder(ASD),and to investigate its possible mechanism.Methods Twenty pregnant rats were taken,in which 16 rats were randomly selected and given liquid prepared with feces of children with ASD of heart-spleen deficiency syndrome bygavage,and the remaining 4 rats were selected as normal control group and giv-en equal volume of normal saline by gavage,all the rats were continuously treated until 21 days after delivery.The success-fully modeled offspring rats with ASD of heart-spleen deficiency type were screened according to three-box social interaction test combined with their body weights,food intake,feces,activity,and appearance.Forty 21-day-old rats were randomly selected from the successfully modeled offspring rats,and randomly divided into model group,Bifidobacterium bifidum group,and low,medium,and high dose groups of Guipi Decoction,with 8 rats in each group,and 8 rats from the offspring rats of the normal control group were selected as the normal group.The Bifidobacterium group was given Bifidobacterium 0.45 g/kg by gavage,and the low,medium and high dose groups of Guipi Decoction were respectively given Guipi Decoc-tion 2.2 g/kg,4.4 g/kg,8.8 g/kg by gavage,the normal group and model group were given equal volume of normal saline by gavage,all once daily,continuously treated for 14 days.The body weights of the rats were recorded at the 7th and 14th day of gavage,their social ability and social novelty were assessed by three-box social test,the content of glutamate in hipp-ocampal tissues of the rats were detected by colorimetric assay,the protein expressions of N-methyl-D-aspartate receptor 2B subunit(NR2B),phosphatase and tension homolog(PTEN),PI3K,Akt,mTOR,p70 ribosomal protein S6 kinase(p70S6K)in hippocampal tissues of the rats were detected by Western blot,the mRNA expression of NR2B,PTEN,P13K,AKT,mTOR and P70S6K in hippocampal tissues of the rats were detected by RT-qPCR.Results Compared with the normal group,the body weight of the rats in the model group was significantly decreased,and their contact time with empty cages was significantly increased while the contact time with unfamiliar rats was significantly decreased(all P＜0.05);the content of glutamate in hippocampal tissues and relative protein and mRNA expressions of NR2B,PI3K,Akt,mTOR,p70S6K in hippocampal tissues were significantly increased(all P＜0.05),and the relative protein and mRNA expressions of PTEN in hippocampal tissues were significantly decreased(all P＜0.05).Compared with the model group,the body weights of the rats in the bifidobacterium group and low,high and medium dose groups of Guipi Decoction were significantly increased,and their contact time with empty cages was significantly decreased while the contact time with unfamiliar rats was significantly increased(all P＜0.05);the contents of glutamate in hippocampal tissues and relative protein and mRNA expressions of NR2B,PI3K,Akt,mTOR,p70S6K in hippocampal tissues were significantly decreased(all P＜0.05),and the relative protein and mRNA expressions of PTEN in hippocampal tissues were significantly increased(all P＜0.05).The body weight and relative protein and mRNA expressions of PTEN in hippocampal tissues of the rats in the midi-um and high dose groups of Guipi Decoction were significantly higher than those in the Bifidobacterium group(all P＜0.05),and the content of glutamate in hippocampal tissues and relative protein and mRNA expressions of NR2B,PI3K,mTOR,and p70S6K in hippocampal tissues were significantly lower than those in the Bifidobacterium group(all P＜0.05).Conclusion Guipi Decoction can effectively improve the social ability and social novelty of rats with ASD of heart-spleen deficiency type,the mechanism of action may be related to down-regulating the content of glutamate and its binding to NR2B receptor,thus to inhibite the activation of PI3K/AKT/mTOR signaling pathway.
autism spectrum disorderGuipi Decoctionheart and spleen deficiency syndromeglutamic acidphos-phatidylinositol 3-kinaseprotein Kinase Bmammalian target of rapamycin
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