The role of neurofilament light chain(NEFL)and neuritin(NRN1)in neuronal apoptosis induced by diacetylmorphine
Objective To investigate the role of neurofilament light chain(NEFL)and neuritin(NRN1)in neuronal apoptosis caused by diacetylmorphine.Methods A diacetylmorphine addicted SD rat model was established and divided into control group and model group,and the pathological changes in brain tissue,the number of apoptotic cells and the protein expression of apoptosis-related factors were observed in each group;Western blot was used to detect the protein expression of NEFL,NRN1,Bax and Bcl-2 in the brain tissues of the rats in each group.Rat primary cerebellar granule cells were transfected using NEFL lentivirus,and the transfection efficiency was detected using Western blot technique,and divided into NC group,model group and sh-NEFL+M group,and diacetylmorphine was used to intervene in model group and sh-NEFL+M group.Cell viability was detected by CCK-8 assay;apoptosis rate were detected by flow cy-tometry;and protein expression of NEFL,NRN1,Bax and Bcl-2 was detected by Western blot.Results HE staining and Hoechst33342 fluorescence staining of brain tissues of addicted SD rats showed that compared with the control group,the brain tissues of the rats in the model group showed obvious sieve-network al-terations;the number of apoptotic cells were significantly increased;the results of Western blot showed that in the model group,the protein expression of NEFL and Bax was increased,and that of NRN1 and Bcl-2 was decreased,and the results were statistically significant(P＜0.05);after diacetylmorphine inter-vened in primary cerebellar granule cells,cell viability was significantly decreased,apoptosis rate was sig-nificantly increased,NEFL and Bax protein expression was significantly increased,and NRN1 and Bcl-2 protein expression was significantly decreased in the model group compared with control group,and the difference was statistically significant(P＜0.05);Compared with the model group,the cell viability of sh-NEFL+M group was significantly increased,apoptosis rate was significantly decreased,NEFL and Bax protein expression was decreased,and NRN1 and Bcl-2 protein expression was increased,and the differ-ence was statistically significant(P＜0.05).Conclusion NEFL and NRN1 play an important role in cere-bellar granule cell apoptosis caused by diacetylmorphine,and decreasing the expression of NEFL can re-duce cerebellar granule cell apoptosis caused by diacetylmorphine.
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