The Protective Effect And Mechanism Of Taohongsiwu Decoction On Injury Induced By Rotenone In SH-SY5Y Cells

Parkinson’s disease(PD),also known as tremor paralysis,is a chronic degenerative disease of central nervous system pathology caused by lsoextrapyramidal dysfunction.The main manifestations is the brain and(or)spinal neuronal degeneration,loss,and accompanied by the death of a large number of nerve cells.So far,the pathogenesis of the disease has not been fully elucidated.Studies have shown that oxidative stress is closely related to PD.Taohongsiwu Decoction(THSWT),a representative prescription of blood stasis and nourishing blood,is widely used in clinic and commonly used in many kinds of diseases caused by blood stasis.In recent years,the study found that Taohongsiwu Decoction and main components can play an important effect on oxidative damage and neuroprotection.In this study,we used rotenone-induced SH-SY5 Y cell injury model to establish the Parkinsonian model for in vitro experiments.When the cells were adhered to 70-80%,MTT method was used to carry out the establishment of rotenone damage model,the screening of the safety concentration range and the screening of protective effect of Taohongsiwu Decoction as well as main components(hydroxysafflor yellow A,fumalic acid,amygdalin,catalpol and paeoniflorin);Inverted microscope was used to observe the morphological changes of cells;Elisa experiment was used to compare the formation of α-synuclein oligomers before and after cell injury;JC-1 mitochondrial membrane potential assay was used to observe the change of mitochondrial membrane potential Δψ before and after theintervention.In addition,to explore the mechanism of neuroprotective effect of Taohongsiwu Decoction and its active ingredients,western blot was used to detect the changes of oxidative stress and apoptosis-related protein expression before and after the intervention,including Bax,bcl-2,ERK1/2,p-ERK,caspase-3,p53.The results showed that,in the range of the safe dose,the cell viabilities of SH-SY5 Y were increased in a dose-dependent manner treated with Taohongsiwu Decoction,hydroxysafflor yellow A(HSY),catalpol and paeoniflorin,etc(P < 0.01).Under the inverted microscope,the cells became round,and the synapses were short and less.In the rotenone-induced PD model,the formation of α-synuclein oligomers was significantly increased(P <0.001),and Taohongsiwu Decoction could inhibit the formation of α-synuclein oligomers(P <0.05).Taohongsiwu Decoction and HSY,ferulic acid,amygdalin,catalpol and paeoniflorin can reverse the phenomenon of Δψ decrease caused by rotenone.Western blot analysis showed that Taohongsiwu Decoction,HSY and paeoniflorin down-regulated the levels of Bax/Bcl-2,caspase-3 and p53 and up-regulated the levels of p-ERK/ERK to antagonize rotenone-induced apoptosis.While the ferulic acid and amygdalin did not significantly reverse the effect of p53 and catalpol on had no significant effect on p-ERK/ERK.In summary,the experiments have shown that Taohongsiwu Decoction has a positive effect on neuroprotective effects.In addition,the in vitro experiments also found that the main components of Taohong Siwu Decoction-HSY,ferulic acid,amygdalin,catalpol and paeoniflorin also have the role of protection of neurons.This may be the result of oxidative stress,mitochondrial function and apoptosis,which may be mediated by ERK pathway(except for catalpol).