Dipsacus asper Treats Alzheimer's Disease in Caenorhabditis elegans by Regulating PPARα/TFEB Pathway
10.13422/j.cnki.syfjx.20240617
- VernacularTitle:基于PPARα/TFEB的川续断干预阿尔茨海默病秀丽隐杆线虫模型作用机制
- Author:
Mengmeng WANG
1
;
Jianping ZHAO
1
;
Limin WU
1
;
Shuang CHU
1
;
Yanli HUANG
1
;
Zhenghao CUI
1
;
Yiran SUN
1
;
Pan WANG
1
;
Hui WANG
1
;
Zhenqiang ZHANG
1
;
Zhishen XIE
1
Author Information
1. Academy of Chinese Medicine Sciences， Henan University of Chinese Medicine， Zhengzhou 450046， China
- Publication Type:Journal Article
- Keywords:
Dipsacus asper;
Alzheimer's disease;
Caenorhabditis elegans;
peroxisome proliferator-activated receptor α （PPARα）;
transcription factor EB （TFEB）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(5):104-114
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the anti-Alzheimer's disease （AD） effect of Dipsacus asper（DA） in the Caenorhabditis elegans model， and decipher the underlying mechanism via the peroxisome proliferator-activated receptor α （PPARα）/transcription factor EB （TFEB） pathway. MethodsFirst， transgenic AD C. elegans individuals were assigned into the blank control， model， positive control （WY14643， 20 µmol·L-1）， and low-， medium-， and high-dose （100， 200， and 400 mg·L-1， respectively） DA groups. The amyloid β-42 （Aβ42） formation in the muscle cells， the paralysis time， and the deposition of amyloid β-protein （Aβ） in the head were detected. The lysosomal autophagy in the BV2 cell model was examined by Rluc-LC3wt/G120A. The expression levels of lysosomal autophagy-related proteins LC3Ⅱ， LC3I， LAMP2， and TFEB were detected by Western blot. Real-time quantitative polymerase chain reaction （Real-time PCR） was employed to determine the mRNA levels of autophagy-related genes beclin1 and Atg5 and lysosome-related genes LAMP2 and CLN2 downstream of PPARα/TFEB. A reporter gene assay was used to detect the transcriptional activities of PPARα and TFEB. Immunofluorescence was used to detect the fluorescence intensity of PPARα， and the active components of the ethanol extract of DA were identified by UPLC-MS. RCSB PDB， Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and Autodock were used to analyze the binding between the active components and PPARα-ligand-binding domain （LBD）. ResultsCompared with the model group， the positive control group and 200 and 400 mg·L-1 DA groups showed prolonged paralysis time （P<0.05）， and all the treatment groups showed decreased Aβ deposition in the head （P<0.01）. DA within the concentration range of 50-500 mg·L-1 did not affect the viability of BV2 cells. In addition， DA enhanced the autophagy flux （P<0.05）， up-regulated the mRNA levels of beclin1， Atg5， LAMP2， and CLN2 （P<0.05， P<0.01）， promoted the nuclear translocation of TFEB （P<0.05）， increased LAMP2 expression and autophagy flux （P<0.05， P<0.01）， and enhanced the transcriptional activities of PPARα and TFEB （P<0.01）. The positive control group and 200 and 400 mg·L-1 DA groups showed enhanced fluorescence intensity of PPARα in the BV2 nucleus （P<0.01）. UPLC-MS detected nine known compounds of DA， from which 8 active components of DA were screened out. The docking results suggested that a variety of components in DA could bind to PPARα-LBD and form stable hydrogen bonds. ConclusionDA may reduce the pathological changes in AD by regulating the PPARα-TFEB pathway.
