The Effect Of Hsa＿circ＿0081596 On FIS1 Expression In Oxygen-glucose-deprivation Induced SK-N-SH Cell Ischemic Injury

Results: qRT-PCR Results:Compared with normal control group(group N),The level of hsa＿circ＿0081596 and FIS1 mRNA in group O and I after OGD/R was up-regulated(P<0.05),the level of hsa＿circ＿0081596 in group S was down-regulated(P<0.05)and the level of FIS1 mRNA in group S was up-regulated;Compared with group O,the level of hsa＿circ＿0081596 and FIS1 mRNA in group S was down-regulated,but there was no significant difference in the level of FIS1 mRNA.the level of hsa＿circ＿0081596 and FIS1 mRNA in group I was up-regulated,but there was no significant difference between the two groups.Objective: Using the cultured human neuroblastoma cells(SK-N-SH),the Oxygen-Glucose-Deprivation(OGD)/Reperfusion(OGD/R)model was established to explore the effect of hsa＿circ＿0081596 on FIS1 expression in Oxygen-Glucose-Deprivation(OGD)induced SK-N-SH cell ischemic injury,and to provide experimental basis for the diagnosis and treatment of cerebral ischemia-reperfusion injury by circular RNAs(circ RNAs).Method: we used human neuroblastoma cell SK-N-SH(within 5 generation)with oxygen-glucose deprivation to imitate ischemic injury in vivo.They were divided into normal control group(Group N),oxygen-glucose deprivation/reperfusion group(Group O),oxygen-glucose deprivation/reperfusion+inhibitor group(Group S)and oxygen-glucose deprivation/reperfusion+inhibitor control group(GroupI).hsa＿circ＿0081596 inhibitor and inhibitor control were transfected into SK-N-SH cell to alter hsa＿circ＿0081596 levels.The expression of hsa＿circ＿0081596 and FIS1 mRNA were determined by Quantitative Real-time PCR.The expression of FIS1 was detected by Western-blot.The extent of cell survival rate was assessed by Cell Counting Kit-8(CCK-8).Cell apoptosis was measured by flow cytometry.SPSS22.0 statistical software was used for data collation and statistical analysis.Conclusion: hsa＿circ＿0081596 can directly regulate the expression of FIS1 rather than regulating FIS1 mRNA,which plays an important role in OGD/R injury by up-regulating the expression level of FIS1 and leading to apoptosis.It suggests that hsa＿circ＿0081596 can be used as a potential therapeutic target for cerebral ischemia-reperfusion injury in circ RNAs level.Western-blot Results:Compared with group N,the expression level of FIS1 in group O and group I was up-regulated(P<0.05)and down-regulated(P<0.05)in group S;Compared with group O,the expression level of FIS1 in group S was significantly down-regulated and up-regulated in group I,but there was no significant difference between the two groups.CCK-8 Results:Compared with group N,the cell survival rate in group O、group S and group I decreased(P<0.05);Compared with group O,The cell survival rate in group S increased and decreased in group I,suggesting that hsa＿circ＿0081596 is involved in SK-N-SH OGD/R injury and low toxicity of si RNA inhibitor control in group I,but because of other factors,the cell survival rate was lower than group S.Flow cytometry Results:Compared with group N,the total cell apoptosis in group O、group S and group I increased(P<0.05),suggesting that apoptosis plays an important role in ischemia-reperfusion injury;Compared with group O,The total apoptosis rate in group S decreased(P<0.05)and increased in group I,but there was no significant difference,suggesting that hsa＿circ＿0081596 involved SK-N-SH cell OGD/R injury may be related to apoptosis.