Protective Effect Of Rutin And Extract Of Bergamot On SH-SY5Y Cells Injured By MPP~+

PD is a neurodegenerative disease and its pathogenesis has not been fully illuminated,but oxidative stress,mitochondrial dysfunction and apoptosis are involved in the pathogenesis of PD.Currently,levodopa(L-dopa)is widely used for the clinical treatment of PD,but it can also cause attenuation of therapeutic effect along with harmful side effects after long time use.Traditional Chinese medicine has more practical experience in the treatment of PD,and the toxicity is relatively smaller,so it is of great significance to study the active components of traditional Chinese medicine.Rutin and extract of Bergamot play an active role in antioxidant and neuroprotection.In conclusion,this experiment explores the protective effect of rutin and extract of bergamot on MPP+-induced SH-SY5 Y cells and provides experimental basis for the development of new drug for PD treatment.The main contents and results are as follows:1.Protective effect of rutin on SH-SY5 Y cells injured by MPP+Methods: The injured model of SH-SY5 Y cells induced by MPP+ was established,and the experiment was divided into control group,MPP+ model group and rutin groups.The model group was treated with 1 mmol · L-1 MPP+,rutin groups were pretreated with different concentrations of rutin(50,100,200,300 mg·L-1)4 h prior to addition of MPP+.After 48 h,cell viability was detected by MTT method,Hoechst 33342 staining was used to observe the changes of nucleus morphology.ROS level was measured by DCFH-DA fluorescence probe,and the mitochondrial membrane potential was detected by JC-1 staining and flow cytometry.the levels of cytochrome C(cytochrome c,Cyt-c)and phosphorylated ERK1/2 protein(P-ERK1/2)were detected by Western blot analysis.Results: Compared with the control group,the cell viability of the model group decreased significantly,the shrunk cell body and nuclear condensation were also observed.ROS level was increased,and the mitochondrial membrane potential was decreased,Cyt-c level increased and phosphorylation ERK1 /2 level was reduced.Compared with the model group,rutin groups(50,100,200,300 mg·L-1)could significantly improve the viability of SH-SY5 Y cells,ameliorated the cell morphology,inhibited the increasement of ROS and restore the membrane potential,Cyt-c level was reduced and phosphorylation ERK1/2 level was increased.Conclusion: Rutin has a significant protective effect on SH-SY5 Y cells injured by MPP+,which may be involved in anti-oxidant activity and ERK activation pathway.2.Protective effect of extract of Bergamot on SH-SY5 Y cells injured by MPP+Methods: The dried Bergamot is crushed,screened,ultrasonic,soaked,and concentrated to obtain the extract,It was placed in the Da 101 macroporous adsorbent resin to separate the components which was the samples of the extract of Bergamot.the experiment was divided into control group,MPP+ injured model group,and EB groups(10,30,60 mg·L-1).following content 1 for the experimental procedures,detected cell viability,observed the changes of nucleus morphology,measured ROS and detected the mitochondrial membrane potential.Results: Compared with the control group,the cell viability of the model group decreased significantly,the shrunk cell body and nuclear condensation were also observed.ROS level in the cells increased obviously,and the mitochondrial membrane potential decreased obviously.Compared with the model group,EB groups(10,30,60 mg·L-1)could significantly improve the cell viability of SH-SY5 Y cells,ameliorated the cell morphology,inhibite the increasement of ROS andrestore the membrane potential,.Conclusion: The extract of Bergamot can protect SH-SY5 Y cells injured by MPP+,but the mechanism need to be clarified.