Effects Of Sema3F On CREBBP Transcription And Protein Expression In Primary Rat Hippocampal Neurons

Objective The aim of this study is to investigate the effect of Sema3F on the transcription and protein expression of CREBBP in primary rat hippocampal neurons,in order to better understand the role of CREBBP in the growth and development of neuronal axons,and to explore the correlation between it and the pathogenesis of epilepsy.Methods 1.Experimental animals selected Sprague-Dawley(SD)rats within 24 hours of neonatal birth,first successfully isolated bilateral hippocampalus in an ultra-clean table under sterile conditions,and performed in vitro culture of primary rat hippocampal neurons in a37℃ CO2 thermostatic cell incubator.2.To confirm the successful establishment of primary rat hippocampal neuron system in vitro,the morphology and axonal growth of hippocampal neurons cultured for 72 hours were observed under inverted phase contrast microscope;3.The successfully cultured and established in vitro primary rat hippocampal neuronal system was divided into experimental group and control group,the experimental group was added with a final concentration of 10ng/ml Sema3 F,while the control group was added with fetal bovine serum of the same concentration,and samples from the above two groups were collected for a total of 4 time points of 0 minutes,5 minutes,15 minutes and 30 minutes.4.Use Real-time PCR to detect the expression of CREBBP m RNA in each sample,and use Western blot method to detect the expression of CREBBP protein in each sample;5.Use SPSS27.0statistical software to statistically analyze the data,the measurement data obey the normal distribution,use the mean ± standard deviation()to express the obtained data,use repeated measurement ANOVA,use LSD-t test to compare the difference between different time points,when the P value < 0.05,indicating that the difference is statistically significant.Results(1)In the primary rat hippocampal neuronal culture system,the expression of CREBBP m RNA in the experimental group decreased at 5 minutes(0.84±0.10),increased at 5minutes(0.93±0.07),and continued to increase at 30 minutes,and was statistically significant at 5 minutes and 15 minutes,while there was no significant change in CREBBP m RNA expression in the control group;(2)In the primary rat hippocampal neuronal culture system,the expression of CREBBP protein in the experimental group decreased at 5 minutes(0.33±0.10),increased at 5 minutes(0.58±0.07),and continued to increase at 30 minutes(0.76±0.14),and there were statistically significant in 5 minutes,15 minutes and 30 minutes,while the expression of CREBBP protein in the control group also did not change significantly.Conclusion(1)Sema3F can change the expression of CREBBP mRNA in primary rat hippocampal neurons,which first decreased and then increased.(2)Sema3F could change the expression of CREBBP protein in primary rat hippocampal neurons,which decreased first and then increased with time.