The Protective Effect Of 3-Hydroxybutyrate Methyl Ester In The Models Of Parkinson’s Disease Induced By Rotenone

BackgroundParkinson’s disease(PD)is the second most common neurodegenerative disease,which is mainly characterized by the destruction of dopaminergic neurons in the substantia nigra compacta.The study found that the pathogenesis of PD may be related to oxidative stress and mitochondrial dysfunction.Metallothionein 2(MT2)could remove oxygen,which is considered to against oxidative stress.At present,the treatment of PD is mainly to supplement the dopamine so that motor symptoms be alleviated.Thus,it is urgent to find a new drug to prevent the occurrence and development of PD.Researchers have found that3-hydroxybutyrate methyl ester(HBME)enhanced the expression of neurotransmitters in brain of rats and improve the cognitive function of the rats.In this condition,we suppose if HBME can adjust the expression of MT2 and play a role of neuroprotection in PD.ObjectiveTo explore the protective effect and the potential mechanism of HBME about Parkinson’s disease induced by rotenone.Method(1)In vitro: SH-SY5 Y cells were divided into three groups: control group,rotenone group and administration group(HBME of 5 m M,10 m M,30 m M).HBME were added prior to rotenone 24 h in administration group and incubated with 0.5μM rotenone for another 24 h subsequently.The viability of SH-SY5 Y cells in different groups were detected by MTT.The intracellular reactive oxygen species(ROS)level and mitochondrial membrane potential(MMP)were observed via DCFH-DA and JC-1 staining under a fluorescence microscopy or flow cytometry.7-AAD /Annexin V-FITC staining were used to test the early apoptosis rate by flow cytometry.Then,MT2 m RNA were tested by qRT-PCR.(2)In vivo: The NIH mice were randomly divided into 3 groups: control group,rotenone group and administration group.The substantia nigra pars compacta(SNc)of control mice were injected into 1μL dimethyl sulfoxide(DMSO)and 0.5μg rotenone were injected into rotenone mice.The HBME(40 mg/kg/d)were supplied for administration mice by oral after the rotenone and lasted 6 weeks.Rotations induced by apomorphine were tested and open field test,pole test,traction test were operated to evaluate the motor function of the mice.The mice were sacrificed after 6 weeks,the number of tyrosine hydroxylase(TH)positive cells in SNc were detected by immunohistochemical staining and the content of neurotransmitter dopamine(DA)were analyzed by high performance liquid chromatography with fluorescence detection(HPLC-FD).Results(1)Compared with the control group,the rotenone decreased the viability of SH-SY5 Y cells and induced apoptosis,the levels of ROS and the MMP were reduced by rotenone.HBME improved and reversed the damage induced by rotenone.In addition,HBME significantly up-regulated the expression of MT2 m RNA.(2)There were no rotation behavior in control mice but the rotational frequency were increased evidently in rotenone mice.Contrasted with control group,the free movement became disorder,the climbing time were prolonged and the time of traction were shortened in rotenone mice.After 6 weeks treatment with HBME,the motor function defict of the mice were improved.In addition,HBME reversed the decrease of the expression of TH and the synthesis of neurotransmitter DA induced by rotenone,respectively.Conclusion HBME could attenuate the damage induced by rotenone in PD models of cells and mice,which may related to the mechanism of up-regulate the expression of MT2 so as to resist oxidative stress and improve mitochondrial dysfunction.