Effect Of Sodium Butyrate On The Expression Of BDNF In Parkinson’s Disease Cell Model

Objective:To observe the protective effect of sodium butyrate on 6-hydroxydopamine(6-OHDA)-induced Parkinson’s disease cell model and its effect on the expression of mBDNF and proBDNF.Methods:6-OHDA at a concentration of 0-200 uM was used to induce PC12 cells in different time,and then the toxic effect of 6-OHDA on PC12 cells was detected by CCK-8 method,and the optimal drug concentration and time of cell modeling were screened.The effects of 6-OHDA at different concentrations and time on cell growth morphology and quantity were observed under microscope.The formation of eosinophilic inclusion bodies was observed by HE staining.The intracellular aggregation of α-synuclein was observed by immunofluorescence assay.Western blotting Western blot was used to detect the expression of intracellular mBDNF in PC12 cells treated with normal PC12 cells and model concentration for 12 h and 24 h.On the basis of successful modeling,the cells were treated with different concentrations of sodium butyrate(NaB)for 1 h,and then co-cultured with 6-OHDA for 24 h.The cell survival of cells treated with sodium butyrate pretreatment was detected by CCK-8 method.The real-time quantitative PCR was used to detect the expression of BDNF and furin(Prorin)mRNA in PC12 cells induced by 6-OHDA.Cellular immunofluorescence was used to observe the expression of TH and BDNF in PC12 cells.Western blot was used to determine the effect of sodium butyrate on the expression of proBDNF,mBDNF,α-synuclein and TH in Parkinson’s disease cell model.Results:1.After 6-OHDA stimulated PC12 cells,it inhibited cell viability and decreased cell viability at a dose(50,100,150,200 uM)and time-dependent manner.2.Under the light microscope,the increase of the concentration of 6-OHDA drug and the prolongation of the action time of PC12 cells under the light microscope,the stronger the growth inhibition,and the cell morphology is full,the contour is irregular,the synapse is stretched,and the shape is rounded and rounded.The synapse becomes shorter or shorter and disappears.3.HE staining found that the cytoplasmic part of the pericytes of the PD cell model showed a circular or elliptical eosinophilic inclusion body,and even some inclusion bodies migrated outside the cell.4.Cellular immunofluorescence showed that there was an inclusion body α-synuclein positive reaction in the cytoplasm of the PD cell model compared with the control group.5.Western blot analysis showed that the induced damage of 6-OHDA showed a time-dependent decrease in mBDNF expression in PC12 cells.6.CCK8 data showed that sodium butyrate could significantly increase the survival rate of PC12 cells induced by 6-OHDA,and it showed a concentration-dependent effect in a certain concentration range.7.qPCR detection confirmed that the BDNF and Furin mRNA were decreased in the model group after 6-OHDA treatment,while the BDNF mRNA was significantly increased after NaB pretreatment,but still lower than the normal level.The level of Furin mRNA in the experimental group treated with NaB was increased compared with the model group,but the difference was not statistically significant(P>0.05).8.The results of immunofluorescence assay showed that the expression of TH and BDNF protein were concentrated in the cytoplasm,and the number of cells expressing protein in the normal group,experimental group and model group decreased in turn.9.Compared with the normal group,the expression of mBDNF protein in the model group was significantly decreased,and the decrease trend was weakened after NaB treatment.The proBDNF protein was slightly lower in the model group than in the normal group,and there was no significant change after NaB treatment compared with the model group(P>0.05).Conclusions:There was an imbalance of proBDNF/mBDNF in the PD model of PC12 cells induced by 6-OHDA,and the proportion of proBDNF increased,and the expression of mBDNF was time-dependent with the induction of 6-OHDA.NaB can influence the expression of BDNF in the model cells from mRNA and protein levels,decrease the proportion of proBDNF/mBDNF protein in PD model cells,increase the survival rate of PD model cells.It was clarified that NaB has a protective effect on6-OHDA-induced PC12 cell Parkinson’s disease in vitro model.