Interference of NIX alleviates spinal cord neurons ischemia-reperfusion injury by inhibiting autophagy
Objective This study aimed to observe whether interference of mitochondrial outer membrane protein NIX could alleviate spinal cord neurons ischemia-reperfusion injury by inhibiting autophagy.Methods Primary spinal cord neurons were isolated and treated with H2O2(200 μmol/mL)to construct the ischemia-reperfusion cell model in vitro.The expression of NIX was knocked down by transfection with shNIX.The apoptosis and proliferation of spinal cord neurons were observed by flow cytometry proliferation of CCK-8 proliferation detection.The transcriptional level of NIX gene was detected by PCR,and the protein expression of NIX,LC3,and Beclin were detected by Western blotting.Results shNIX significantly downregulated the transcription level and protein expression levels of NIX,and the inhibition effect is obvious,which is used for subsequent experimental operations.Flow cytometry proliferation of CCK-8 proliferation detection showed that ischemia-reperfusion injury induced apoptosis of spinal cord neurons with reduced proliferation rate.In addition,ischemia-reperfusion injury promoted the expressions of NIX,LC3,and Beclin.More importantly,shNIX reduced the apoptosis of spinal cord neurons,promoted cell proliferation,as well as reduced the protein expressions of LC3 and Beclin.Conclusion Interference of NIX attenuated ischemia-reperfusion induced spinal cord neurons injury by inhibiting autophagy.
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