Effects Of MiR-143-3p On NRG1 In Alzheimer’s Disease

ObjectiveTo investigate and analyze the effect of mir-143-3p on nerve cell survival in Alzheimer’s disease(Alzheimer’s disease,AD)cell model.Methods(1)Firstly,SH-SY5 Y cells were induced to differentiate into neurons by all trans retinoic acid(Retinoic acid,RA).The morphology of cells was observed by light microscope,and the expression of tubulin βIII was detected by immunofluorescence method.(2)Aβ1-42 was used After inducing differentiation of SH-SY5 Y cells,AD cell model was established,cell viability was detected by CCK-8,neuronal cell death was detected by TUNEL,miR-143-3p expression was detected by quantitative PCR,NRG1 protein expression was detected by Western blot.(3)miR-143-3p inhibitor was transfected into SH-SY5 Y cell line,and SH-SY5 Y cells were treated with Aβ1-42,The m RNA and protein expression of miR-143-3p,NRG1 and related apoptotic protein were detected by quantitative PCR and Western Blot method,the cell viability was detected by CCK-8 method,and the neuronal cell death was detected by TUNEL method.(4)the interaction between miR-143-3p and NRG1 was detected by luciferase reporter gene method,miR-143-3p inhibitor / mimic was transfected in SH-SY5 Y cell line,quantitative PCR and Western blot were used to detect the expression of NRG1 in SH-SY5 Y cells.(5)miR-143-3p inhibitor and NRG1 si RNA were co transfected in SH-SY5 Y cells,and SH-SY5 Y cells were treated with Aβ1-42.CCK-8 was used to detect cell viability,TUNEL was used to detect neuronal celldeath.Results(1)The SH-SY5 Y cells induced by RA showed typical neuron-like synapses,and immunofluorescence test showed increased expression levels of tubulin III.(2)SH-SY5 Y cells were treated with Aβ1-42 to construct an AD cell model.Compared with cells in the control group,miR-143-3p was significantly increased in the AD cell model(P<0.01),and NRG1 was significantly lower(P<0.001).(3)Compared with the control group,the proportion of apoptosis in nerve cell was significantly increased(P<0.001)and the cell survival rate was significantly decreased(P<0.001)in the AD cell model.After targeted inhibition of miR-143-3p expression,the percentage of apoptosis cells induced by Aβ1-42 was significantly decreased(P<0.01),the expression level of NRG1 protein was significantly increased(P<0.01),and the expression levels of lysase Caspase3 and Caspase9 protein were significantly decreased(P<0.001).(4)The double luciferase gene test reports showed that miR-143-3p and NRG1 had targeted binding sites,and NGR1 was the downstream target gene of miR-143-3p.(5)Targeted silencing of NGR1 expression could reverse the effect of miR-143-3p expression inhibition of viability(P<0.01)and promotation of apoptosis(P<0.001)on AD cell.Conclusions1.miR-143-3p was highly expressed in AD model and NRG1 was low expressed in AD model..2.NRG1 is the target gene of miR-143-3p,and miR-143-3p inhibits the survival of neural cells in AD model through NRG1.