Spinosin Inhibits Aβ1-42 Production and Aggregation via Activating Nrf2/HO-1 Pathway
Xiaoying Zhang1, Jinyu Wang1, Guowei Gong2
1Key Laboratory of Active Components of Chinese Medicine Screening and Evaluation, School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China.
Abstract:
The present research work primarily investigated whether spinosin has the potential of improving the pathogenesis of Alzheimer's disease (AD) driven by β-amyloid (Aβ) overproduction through impacting the procession of amyloid precursor protein (APP). Wild type mouse Neuro-2a cells (N2a/WT) and N2a stably expressing human APP695 (N2a/APP695) cells were treated with spinosin for 24 h. The levels of APP protein and secreted enzymes closely related to APP procession were examined by western blot analysis. Oxidative stress related proteins, such as nuclear factor-erythroid 2-related factor 2 (Nrf2), and heme oxygenase-1 (HO-1) were detected by immunofluorescence assay and western blot analysis, respectively. The intracellular reactive oxygen species (ROS) level was analyzed by flow cytometry, the levels of Aβ1-42 were determined by ELISA kit, and Thioflavin T (ThT) assay was used to detect the effect of spinosin on Aβ1-42 aggregation. The results showed that ROS induced the expression of ADAM10 and reduced the expression of BACE1, while spinosin inhibited ROS production by activating Nrf2 and up-regulating the expression of HO-1. Additionally, spinosin reduced Aβ1-42 production by impacting the procession of APP. In addition, spinosin inhibited the aggregation of Aβ1-42. In conclusion, spinosin reduced Aβ1-42 production by activating the Nrf2/HO-1 pathway in N2a/WT and N2a/ APP695 cells. Therefore, spinosin is expected to be a promising treatment of AD.
Insights
Spinosin shows promise for Alzheimer's disease (AD) treatment by reducing amyloid-beta (Aβ) production and aggregation. It achieves this by activating the Nrf2/HO-1 pathway, mitigating oxidative stress, and impacting amyloid precursor protein (APP) processing.
Area of Science:
- Neuroscience
- Pharmacology
- Biochemistry
Background:
- Alzheimer's disease (AD) pathogenesis is linked to β-amyloid (Aβ) overproduction and processing of amyloid precursor protein (APP).
- Oxidative stress plays a significant role in AD progression, influencing APP processing enzymes like ADAM10 and BACE1.
- Nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) are key regulators of the cellular antioxidant response.
Purpose of the Study:
- To investigate the potential of spinosin in ameliorating Alzheimer's disease (AD) pathogenesis.
- To determine if spinosin affects amyloid precursor protein (APP) processing and subsequent β-amyloid (Aβ) production.
- To elucidate the mechanism by which spinosin might exert its effects, focusing on oxidative stress and the Nrf2/HO-1 pathway.
Main Methods:
- Treatment of wild type mouse Neuro-2a (N2a/WT) and N2a/APP695 cells with spinosin.
- Western blot analysis to quantify APP protein and related processing enzymes (ADAM10, BACE1).
- Immunofluorescence and western blot for oxidative stress markers (Nrf2, HO-1).
- Flow cytometry for intracellular reactive oxygen species (ROS) levels.
- ELISA for Aβ1-42 quantification.
- Thioflavin T (ThT) assay for Aβ1-42 aggregation.
Main Results:
- Spinosin inhibited ROS production by activating the Nrf2 pathway and up-regulating HO-1 expression.
- Spinosin treatment led to reduced expression of BACE1 and increased expression of ADAM10, impacting APP processing.
- Spinosin significantly decreased Aβ1-42 production and inhibited its aggregation.
- The observed effects were consistent in both N2a/WT and N2a/APP695 cell lines.
Conclusions:
- Spinosin effectively reduces Aβ1-42 production and aggregation in neuronal cells.
- The mechanism involves the activation of the Nrf2/HO-1 antioxidant pathway, which counteracts oxidative stress.
- Spinosin's ability to modulate APP processing and reduce Aβ burden suggests its potential as a therapeutic agent for Alzheimer's disease.
