Protective Effect of Danggui Shaoyaosan-contained Serum on Aβ1-40-injured PC12 Cells via Regulating UPP and Its Mechanism
10.13422/j.cnki.syfjx.20211207
- VernacularTitle:当归芍药散含药血清调控UPP途径对Aβ1-40损伤PC12细胞的保护作用及机制
- Author:
Yunhui CHEN
1
;
Jun XIA
1
;
Xinglong LIU
1
;
Wenying HUAI
1
;
Dan LIU
2
;
Tiane ZHANG
1
;
Yongmei XIE
2
;
Yu YOU
1
;
Wen YUE
3
;
Songqi TANG
3
;
Wei PENG
1
Author Information
1. Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
2. Sichuan University，Chengdu 610041，China
3. Hainan Medical University，Haikou 570102，China
- Publication Type:Journal Article
- Keywords:
Danggui Shaoyaosan;
Alzheimer's disease （AD）;
ubiquitin-proteasome pathway （UPP）;
rat adrenal pheochromocytoma （PC12） cells
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(24):17-25
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the protective effect of Danggui Shaoyaosan （DSS）-contained serum on β-amyloid （Aβ）1-40-injured rat adrenal pheochromocytoma PC12 cells and its mechanism in regulating ubiquitin-proteasome pathway （UPP）. MethodAβ1-40 was used to intervene PC12 cells to prepare the cell models of Alzheimer's disease （AD）， and the experiment was divided into the blank， model， and DSS-contained serum high， medium， and low-dose groups （10%， 5%， and 2.5%）. Cell viability and apoptosis were detected using cell counting kit-8 （CCK-8） method and flow cytometry， respectively. The content of Aβ and p-Tau protein was determined by enzyme-linked immunosorbent assay （ELISA）. The ubiquitin （Ub）， ubiquitin ligase E3 （E3）， 26S proteasome， ubiquitin carboxyl terminal hydrolase1 （UCHL1）， and UCHL3 protein expressions of UPP were displayed using immunofluorescence cytochemistry （ICC）， and the mRNA and protein expression levels of Ub， E3-parkin， 26S， UCHL1， and UCHL3 were determined by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultThe data of the CCK8 experiment verified that 5 μmol·L-1 and 48 hours were the optimal conditions for modeling Aβ1-40-injured PC12 cells. As compared with the blank group， the cell viability rate in the model group decreased （P<0.05） with an increased apoptosis rate （P<0.05）， the content of Aβ and p-Tau contents was elevated （P<0.05）， the mRNA and protein expression levels of Ub increased， and the mRNA and protein expression levels of 26S， E3， and UCHL1+3 decreased （P<0.05）. As compared with the model group， the cell viability rate in the DSS-contained medium-dose group increased （P<0.05）， whereas the apoptosis rate in each DSS-contained group decreased （P<0.05）. The content of Aβ in each DDS-contained group decreased （P<0.05）， and the content of p-Tau in the DDS-contained high and medium-dose groups decreased （P<0.05）. The mRNA expression level of Ub decreased， and that of 26S increased in each DDS-contained group （P<0.05）. The mRNA expression level of UCHL1 in the DDS-contained medium-dose group increased （P<0.05）， and the mRNA expression levels of E3 and UCHL 3 in the DDS-contained high and medium-dose groups increased （P<0.05）. The protein expression level of Ub in each DDS-contained group decreased， and the protein expression levels of 26S， E3， and UCHL1+3 in the DDS high and medium-dose groups increased. The DSS-contained serum medium-dose group exerted the optimal effect. ConclusionDSS-contained serum can increase cell viability rate， reduce cell apoptosis rate， eliminate Aβ and p-Tau protein deposits， and exert protective effects on Aβ1-40-injured PC12 cells. Its mechanism may involve UPP via decreasing the expression of Ub and increasing that of 26S， E3， UCHL1， and UCHL3.
