ABSTRACT
OBJECTIVE To investigate the autophagic effect of the compounds from the Chinese medicinal herbs, Radix polygalae as a potential neuroprotective agent that enhances the clearance of mutant Huntingtin and α- synuclein in PC- 12 cells. METHODS Radix polygalae was extracted with 75% ethanol using refluxing method, and its quality was assayed by UHPLC-TOF-MS. The autophagic effect of Radix polygalae extract, and its major components including polygalacic acid, senegenin and onjisaponin B were investigated using the green fluorescent protein-light chain 3 (GFP-LC3) autophagy detection and Western blot platforms for detecting the autophagic markers, GFP-LC3 puncta formation and LC3 II expression. The degradation of A53T α- synuclein and the inhibition ofα-synuclein oligo merization related to the Parkinson disease were assayed using Western blot and flow cytometer analysis, respectively. While the cytotoxicity and the degradation of the mutant Huntingtin associated with the Huntingtin disease were investigated using MTT method and flow cytometer analysis. RESULTS Radix polygalae ethanol extract and onjisaponin B improved the GFP-LC3 puncta formation and expression of LC3Ⅱ with time and dose manner, and this induction was activated via AMPK-mTOR and Atg 7 gene pathway. Furthermore, the clearance of α-synuclein and mutant Huntingtin was enhanced via autophagy induction with the treatment of Radix polygalae ethanol extract and onjisaponin B. CONCLUSION Findings in the current study provide detailed insights into the protective mechanism of a novel autophagy inducer, onjisaponin B, which is valuable for further investigation as a new candidate agent for modulating neurodegenerative disorders through the reduction of toxicity and clearance of mutant proteins in the cellular level.
ABSTRACT
OBJECTIVE To investigate the effect of neuroprotective effect of lychee seed saponins (LSS) in BV-2. METHODS Aβ1-42 induced BV-2 cells were incubated with LSS for 12 h, the content of the inflammatory factors such as IL-1β, TNF-α, COX-2 and iNOS in the supernatant of BV-2 cell were measured by ELISA. The detection of the mRNA levels and the protein expression of the inflammatory factors including IL-1β, TNF-α, COX-2 and iNOS using real-time PCR and Western blotting, respectively. RESULTS The level of IL-1β, COX-2 and iNOS significantly increased with the treatment of Aβ1-42, and 0.117 mg·L-1-0.469 mg·L-1 LSS can inhibit these increased level. CONCLUSION LSS conferred neuroprotection via inhibiting the inflammatory factors expression.