The Protective Effect And Its Mechanism Of Simvastatin In Parkinson's Disease Model

BackgroundParkinson's disease is the second most frequent neurodegenerative disorder,which is secondly to AD,it is mainly manifested as bradykinesia,postural instability,rigidity and resting tremor,resting tremor and other sports symptoms.The incidence of PD over the age of 65 is 1%-2%around the world.The levodopa replacement therapy is mainly traditional strategy for PD treatment,although it can control the clinical symptoms of patients temporarily,it can not prevent the progression of the disease.The study of PD is still a hot topic in medical research nowadays,and oxidative stress injury is gradually regarded as one of the important mechanisms leading to the pathogenesis of PD.Simvastatin,as a lipid-lowering drug,inhibits HMG-CoA reductase activity and reduce oxidative stress?induced injury to vascular.Recent studies have shown that statins may be beneficial for PD by regulating the signaling pathways associated with neurodegeneration.Among all kinds of statins,simvastatin is the most likely through the blood-brain barrier,it can effectively play a neuroprotective effect in the brain.Objectives(1)To define that 6-OHDA mediated nerve injury by oxidative damage in PD cell model and animal model;(2)To elucidate the protective effect of simvastatin on 6-OHDA-induced PD cell model and animal model;(3)Explain the antioxidant mechanisms of simvastatin associated with neuroprotective effect in PD cell model and animal model.MethodsIn this study,6-OHDA?treated cells and mice were employed as PD model to study the protective effect of simvastatin on PD and its related antioxidant mechanisms.The main research methods employed d include:(1)CCK-8 kit was employed to detect the changes of cell viability;(2)Hoechst 33342 staining was employed to detect the apoptosis of the cells;(3)Western Blot was employed to detect the expression of iNOS,cleaved caspase 3,Ho-1,PGC-1?,p38 MAPK,p-p38 MAPK and gp91 pHox in the cells.(4)Immunocytlfluorescence was employed to detect the nuclear translocation of NF-?B and the membrane metastasis of p-47 pHOX.(5)We detected intracellular ROS content using DCFH-DA fluorescent dye.(6)SOD detection kit was used to detect the content of SOD in midbrain.(7)The inflammatory factors in mice serum were detected by ELISA.Results(1)6-OHDA could induce the expression of cleaved caspase 3 in SH-SY5Y cells(180.1±10.4%compared to control),and lead to the decrease of Bel-2/Bax(65.0±4.0%compared to control)and cell viability(45.0±0.6%compared to control).Simvastatin could increase the viability of 6-OHDA-treated SH-SY5Y(59.6±5.8%compared to control),increase the ratio of Bcl-2/Bax(96.0±9.3%compared to control),inhibit the expression of cleaved caspase 3(126.5±10.3%compared to control);(2)6-OHDA could induce iNOS expression(173.1±39.7%compared to control)and ROS production in SH-SY5Y cells;Simvastatin reduced iNOS expression(78.6±7.3%compared to control)and ROS production in 6-OHDA-treated SH-SY5Y cells;(3)6-OHDA could activate NADPH enzyme,increase the expression of NADPH enzyme subunit gp91PHOX(136.0±5.7%compared to control)and promote membrane metastasis of NADPH enzyme subunit p47PHOX,and increase the level of p-p38 MAPK(191.7±21.5%compared to control)and the translocation of NF-?B;Simvastatin could reduce the expression of NADPH enzyme subunit gp91PHOX(94.2±4.2%compared to control)and inhibit the membrane metastasis of the NADPH enzyme subunit p47PHOX in 6-OHDA-induced SH-SY5Y cells;Simvastatin could also inhibit the activation of p38 MAPK(123.1±9.0%compared to control)and the translocation of NF-?B;(4)Simvastatin could increase the expression of antioxidant protein HO-1(245.0±13.0%compared to control)and PGC-1?(159.2±15.4%compared to control)in 6-OHDA-treated SH-SY5 Y cells;(5)The frequency of right forelimb use and apomorphine-induced rotation increased in 6-OHDA-injected mice;Simvastatin could decrease the frequency and rotation.(6)The content of IL-1? in mice serum increased,expression of SOD,GCLM,PGC-1?in midbrain decreased in 6-OHDA-injected mice;After simvastatin treatment,the level of IL-1? decreased,and the expression of SOD(75.3±9.1%compared to control),GCLM(71.1±7.6%compared to control),PGC-1?(89.9±3.8%compared to control)in midbrain increasedConclusions(1)6-OHDA could activate NADPH oxidase,p38 MARK and NF-?B,produce oxidative stress factor,induce oxidative damage and cell death,.Simvastatin can inhibit the production of reactive oxygen species by inhibiting the activity of NADPH oxidase and p38 MARK,and NF-KB,protect cells from damage.(2)6-OHDA can harm the behavior of PD mice,increase the secretion of inflammatory factor in peripheral blood and destroy the antioxidant defense system,damage the antioxidant defense system,Simvastatin can decrease the oxidative damage injury in PD mice and cells through increasing the expression of antioxidant proteins...