The Mechanism Of Rosmarinic Acid Reducing SH-SY5Y Parkinson’s Disease Cell Model Damage By Regulating Abl

Background: Rosmarinic acid(RA)is a natural polyphenol compound with antioxidant properties.Neurological diseases are now the main source of global disability.The fastest-growing neurological disease in the world is Parkinson’s disease(PD).The demand for the development of accurate dietary compounds using natural plants to prevent and treat Parkinson’s disease is increasing.This study aims to use the SH-SY5 Y Parkinson’s cell model induced by rotenone to study the new potential mechanism of action of rosmarinic acid,providing scientific support for promoting practical applications.Method : Cell viability was measured by MTT method,cell morphology was observed by inverted microscope,apoptosis was detected by Annexin V-PI double staining,reactive oxygen species(ROS)was detected by DCFH-DA probe,ATP content was detected by kit,mitochondrial membrane potential was detected by JC-1 probe,and Western Blot assay was performed α-Synuclein、P-Akt、P-Ampk、P-Tau、PGC-1α、Phosphorylation and expression of proteins such as P-Abl,P-Crk,and GDNF.At the same time,we will construct a neuron-glial cell culture system to explore the protective effect of rosmarinic acid on glial cells and its supporting effect on neurons.Results: Rosmarinic acid exhibits dose-dependent neuroprotective effects in the SH-SY5 Y cell model of Parkinson’s disease induced by rotenone,100 μM rosmarinic acid decreased by 50 μM rotenone reduced cell viability,improved cell apoptosis,restored normal cell morphology,and reduced LDH levels in cell suspensions;100 μM rosmarinic acid reduced by Elevated levels of reactive oxygen species caused by 50 μM rotenone;In terms of mitochondrial function,rosmarinic acid restored the decrease of membrane potential and ATP level caused by 50 μM rotenone;At the protein level,rosmarinic acid not only decreased α-Synuclein protein levels,Bax protein levels,and Abl Ser473 phosphorylation also increase Akt Ser473 phosphorylation and Bcl-2 protein levels.Further research results indicate that the neuroprotective effect of rosmarinic acid is partially due to its inhibition of Abl tyrosine kinase.Rosmarinic acid treatment inhibited rotenone induced hyperphosphorylation of Abl Y412 and Crk II Y221,and the mechanism involved decreased ROS level,increased membrane potential,increased ATP level,decreased α-synuclein protein,decreased phosphorylation of Tau Ser404,increased phosphorylation of Akt Ser473,increased phosphorylation of AMPK Thr172,increased levels of Mfn2 protein,increased levels of OPA1 protein,and PGC-1α protein level increases.In addition,rosmarinic acid also showed neuroprotective effect in the rotenone induced U251 cell model,which could increase the level of glial neurotrophic factor(GDNF)in glial cells.Through the constructed neuron glial cell culture system,it was found that GDNF protected the SH-SY5 Y cell vitality reduced by rotenone.