Study On The Neurobehavioral And Mitochondrial Damage Mechanism Of Matrine In Mic

Objective:To study the effects of matrine on neural behavioral changes in mice and the toxic effects of Rat adrenal pheochromcytoma cells(PC 12 cells),and to investigate the mitochondrial mechanism of neuronal toxicity of different concentrations of matrine,in order to provide the reference for the security of matrine in clinical practice.Methods:1.Neural behavioral changes of matrine in mice:60 ICR mice were randomly divided into control group,1Omg/kg and 40 mg/kg matrine group,once a day for 60 days.During the experiment,the physiological behavior of the animals was observed,such as activity,diet,coat color,mental state and so on.The weight of the mice was recorded once a week.Before the first dose and after the last administration,the number of independent activities in mice were detected by a multi-functional autonomous instrument,the time of climbing in each experimental group was determined at the same time.2.Toxic effects of matrine on PC12 cells:PC 12 cells were treated with different concentrations of matrine for 8 h,16 h,24 h and 48 h respectively.The viability of PC 12 cells and the release rate of lactate dehydrogenase(LDH)were determined;morphological changes of PC 12 cells were observed by inverted microscope and Hoechst 33342 fluorescent staining.3.Mitochondrial mechanism of neuronal toxicity of different concentrations of matrine:The activity of superoxide dismutase(SOD)and malondialdehyde(MDA)in PC 12 cells were detected by WST-8 method,the changes of mitochondrial membrane potential(MMP)were detected by JC-1 staining and the apoptosis rate was detected by Annexin-FITC/PI double staining.Western blot was used to detect the levels of caspase 3,Caspase 8 and caspase 9 and Bax,Bcl-2,in order to study the mitochondrial mechanism of neuronal toxicity of different concentrations of matrine.Results:1.The mice were treated with different doses of matrine for 60 days.The control group had normality activity;while the mice in the administration group were curled up,depressed and had less activity.The body weight of mice in each group was measured every 7 days.The results showed that the body weight of each group was increased,the body weight of the mice was significantly lower than that of the control group(P<0.05(P<0.01),suggesting that different doses of matrine can inhibit the central nervous system and change autonomous activities、the balance and coordination ability of mice(P<0.01).2.PC 12 cells were exposed to different concentrations of matrine,the cell viability and the release of LDH were;PC 12 cell morphology were changed and the nucleus appeared to shrink,partially broken and other apoptotic phenomena,showing that different concentrations of matrine on PC 12 cells have a certain toxic effect.3.Exposed to different concentrations of matrine for 24 h,the level of ROS,the content of MDA were increased,SOD activity and mitochondrial membrane potential were decreased,Bcl-2,Caspase 3,Caspase 8 and caspase 9 were decreased,and the expression of Cleaved-caspase 3 was up-regulated,suggesting that different concentrations of matrine can change the oxidative stress in PC 12 cells,resulting in oxidative stress injury,making the cell membrane damage,mitochondrial membrane potential decreased,activated Caspase cascade reaction,induced apoptosis.Conclusion:1.Different doses of matrine can inhibit the central nervous system,change the balance of mice coordination ability,had a certain impact on the neurocognitive function of mice.2.Different concentrations of matrine have a certain toxic effect on PC 12 cells.3.Different concentrations of matrine can change the oxidative stress in PC 12 cells,resulting in oxidative stress injury,making the cell membrane damage,mitochondrial membrane potential decreased,activated Caspase cascade reaction,induced apoptosis.