Effect of Shouwuwan on Apoptosis of Hippocampal Neurons in Aging Rats Induced by D-galactose Based on PI3K/Akt Pathway
10.13422/j.cnki.syfjx.20222137
- VernacularTitle:基于PI3K/Akt通路探讨首乌丸对D-半乳糖衰老大鼠海马神经元凋亡的影响
- Author:
Chao ZHANG
1
;
Xiufeng JIA
1
;
Wanling HUANG
1
;
Yong ZHOU
2
;
Qi TANG
1
;
Gang CHEN
1
Author Information
1. School of Basic Medicine， Hubei University of Chinese Medicine， Wuhan 430065， China
2. Wuhan Hospital of Traditional Chinese Medicine（TCM）， Wuhan 430017， China
- Publication Type:Journal Article
- Keywords:
brain aging;
Shouwuwan;
neuronal apoptosis;
phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathway;
forkhead box protein O3a （FoxO3a）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(22):76-84
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of Shouwuwan on apoptosis of hippocampal neurons in D-galactose-induced aging rats through phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathway. MethodFifty male SD rats of SPF grade were randomly divided into normal group， model group， vitamin E group， and Shouwuwan medium- and high-dose groups. Except the normal group， the other four groups were given D-galactose （120 mg·kg-1） to prepare aging model. Additionally， Shouwuwan was used to intervene in the Shouwuwan medium- and high-dose groups （1.08 and 2.16 g·kg-1， respectively）， and vitamin E group （0.018 g·kg-1） was given vitamin E by gavage. After 6 weeks of modeling， the whole brain and hippocampal tissue were taken and the morphological changes of hippocampal neurons were observed by Nissl staining. The apoptosis of hippocampal cells was detected by in situ end labeling ［TdT-mediated dUTP nick-end labeling （TUNEL）］. The protein expression levels of PI3K， phosphorylated （p）-PI3K， Akt， p-Akt， Caspase-3， forkhead box protein O3a （FoxO3a）， p-FoxO3a， B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax） were detected by Western blot. Real-time fluorescence quantitative PCR （Real-time PCR） was performed to determine the mRNA expression level of FoxO3a in hippocampus. ResultCompared with the conditions in the normal group， the apoptotic cells of hippocampal neurons in the model group were significantly increased， and the apoptosis index was elevated （P<0.01）. Nissl staining of the hippocampal CA1 region showed that hippocampal neurons were lost and sparse， and the number of Nissl bodies was reduced， with pyknosis and deep staining. The relative protein expression of p-PI3K， p-Akt， Caspase-3 and Bax， p-PI3K/PI3K ratio and p-Akt/Akt ratio were all increased （P<0.01）， while the relative protein expression of FoxO3a， p-FoxO3a and Bcl-2， and p-FoxO3a/FoxO3a ratio were decreased （P<0.01）. The mRNA expression of FoxO3a was lowered （P<0.01）. Compared with the conditions in the model group， after treatment with Shouwuwan， the apoptotic cells were markedly reduced， and the apoptosis index of each treatment group was decreased （P<0.01）. Nissl staining of the hippocampal CA1 region demonstrated that the loss of neurons in each treatment group was improved， and Nissl bodies were increased and densely arranged. There was no statistically significant difference in the relative protein expression of PI3K and Akt in each group. In Shouwuwan medium- and high-dose groups， the relative protein expression of p-PI3K， p-Akt， Caspase-3 and Bax， p-PI3K/PI3K ratio and p-Akt/Akt ratio were decreased， while the relative protein expression of FoxO3a andp-FoxO3a， and Bcl-2， and p-FoxO3a/FoxO3a ratio were increased （P<0.01）. The mRNA expression of FoxO3a was up-regulated （P<0.01）. ConclusionShouwuwan could improve the structure of hippocampal neurons， inhibit PI3K/Akt signal pathway， down-regulate Caspase-3 and Bax， activate FoxO3a， and up-regulate Bcl-2 and Bcl-2/Bax ratio， to reduce neuronal apoptosis.
