Liuwei Dihuangwan Reduces Valproic Acid-induced ASD-like Behaviors in Rats by Modulating mTOR-mediated Synaptic Plasticity in Prefrontal Cortex-hippocampus Circuit
10.13422/j.cnki.syfjx.20251419
- VernacularTitle:六味地黄丸通过调控前额叶-海马环路mTOR介导的突触可塑性改善丙戊酸诱导的孤独症模型大鼠行为分析
- Author:
Ji WU
1
;
Niya YIN
1
;
Lei OUYANG
2
;
Siyu HE
1
;
Fangyu PEI
3
;
Ying XIE
4
;
Fan LI
4
;
Qian LEI
4
;
Sha ZHAO
5
;
Di ZHANG
4
;
Qinquan ZHU
4
Author Information
1. The First Clinical School of Chinese Medicine， Hunan University of Chinese Medicine， Changsha 410007，China
2. Guangdong Second Traditional Chinese Medicine Hospital， Guangzhou 510030，China
3. Longgang District People's Hospital， Shenzhen 518000，China
4. The First Hospital of Hunan University of Chinese Medicine， Changsha 410007，China
5. The 922nd Hospital of the Joint Logistics Support Force， Hengyang 421002，China
- Publication Type:Journal Article
- Keywords:
Liuwei Dihuangwan;
autism spectrum disorder;
mammalian target of rapamycin （mTOR） signaling pathway;
synaptic plasticity
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(22):1-11
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effects of Liuwei Dihuangwan on mammalian target of rapamycin （mTOR）-mediated synaptic plasticity in the prefrontal cortex-hippocampus circuit in the rat model of valproic acid （VPA）-induced autism spectrum disorder （ASD） and to explore its potential mechanism for reducing behavioral abnormalities in ASD rats. MethodsThirteen SD pregnant rats were randomized into the VPA group （10 rats） and the control group （3 rats）. On gestational day 13， the VPA group was intraperitoneally injected with VPA solution （600 mg·kg-1）， while the control group was injected with an equal volume of normal saline. Male pups of 21 days old from both groups underwent behavioral tests to screen the ASD model rats （32 rats）. These rats were then randomly assigned to the model （distilled water）， vitamin D （1 480 U·kg-1）， low-dose （0.75 g·kg-1） Liuwei Dihuangwan， and high-dose （1.5 g·kg-1） Liuwei Dihuangwan groups， and normal male pups were selected as the blank group （distilled water）， with 8 rats in each group. Each group was treated with the corresponding drug or an equal volume of distilled water via gavage daily. After drug administration， behavioral tests were conducted again， followed by anesthesia and tissue collection. Nissl staining was carried out to observe the morphological changes of neurons in the prefrontal cortex and hippocampus. Transmission electron microscopy was used to observe changes in hippocampal synaptic structure. Immunohistochemistry and Western blot were employed to determine the expression of mTOR， phosphatidylinositol 3-kinase （PI3K）， and protein kinase B （Akt） in the prefrontal cortex， as well as synaptophysin-1 （SYP-1）， postsynaptic density 95 （PSD95）， and N-methyl-D-aspartate receptors （NMDARs） in the hippocampus. ResultsCompared with the control group， the model group exhibited severe growth retardation and developmental deformities， with abnormal behavioral performance and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus. Compared with the model group， all treatment groups showed reduced behavioral abnormalities and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus， with the most significant effects in the high-dose Liuwei Dihuangwan group. Immunohistochemistry and Western blotting results showed that compared with the control group， the model group exhibited significantly increased protein levels and phosphorylation ratios of p-mTOR， p-Akt， and p-PI3K in the prefrontal cortex （P<0.05， P<0.01）， and decreased protein levels of SYP-1 and PSD95 and increased protein level of NMDARs in the hippocampus （P<0.01）. Compared with those in the model group， the protein levels of p-mTOR， p-Akt， and p-PI3K in the prefrontal cortex decreased in the vitamin D and high-dose Liuwei Dihuangwan groups （P<0.05）， and the p-mTOR/mTOR， p-Akt/Akt， and p-PI3K/PI3K ratios in the prefrontal cortex declined in the vitamin D group （P<0.05）. The high-dose Liuwei Dihuangwan group showed a reduction in p-PI3K/PI3K ratio （P<0.05）. The protein level of SYP-1 and PSD95 in the hippocampus increased in the vitamin D， low-dose Liuwei Dihuangwan， and high-dose Liuwei Dihuangwan groups （P<0.05， P<0.01）， while the protein level of NMDARs decreased in the vitamin D and high-dose Liuwei Dihuangwan groups （P<0.01）. ConclusionLiuwei Dihuangwan significantly reduces VPA-induced ASD-like behaviors and enhances the regeneration and repair of neurons in the prefrontal cortex and synaptic structures in the hippocampus by downregulating of PI3K/Akt/mTOR expression in the prefrontal cortex， upregulating SYP-1 and PSD95 expression in the hippocampus， and inhibiting NMDARs expression.
