The Mechanism Of MiR-486-5p Regulation Of MST4 Expression Mediating Brain Hemorrhage Injury

Objective: To investigate whether mi R-486-5p expression levels are altered in the serum of patients with acute cerebral hemorrhage and to further investigate the effect of mammalian sterile-20-like kinase 4(MST4)expression on cell viability after cerebral hemorrhage injury,and to investigate whether mi R-486-5p targets MST4 to regulate the pathological injury process after cerebral hemorrhage.Materials and METHODS: Bioinformatics was used to screen mi RNA that were differentially expressed and might interact with MST4 after intracerebral hemorrhage.Double luciferase reporter gene was used to detect the targeting relationship between mi R-486-5p and MST4.A total of 30 patients with acute cerebral hemorrhage admitted to the Department of Neurology from April 2021 to July 22,2021 and 30 healthy subjects were collected.Quantitative real-time polymerase chain reaction(q RTPCR)was used to detect the serum mi R-486-5p levels in patients with cerebral hemorrhage and healthy people.The expression level of mi R-486-5p was correlated with the volume of hematoma by calculating the volume of hematoma based on head CT images.Hemin-induced HT-22 cell injury was used to establish an in vitro cerebral hemorrhage model.Quantitative real-time polymerase chain reaction(q RTPCR)was used to detect the expression level of mi R-486-5p in HT-22 cells after bleeding injury and in normal cells.CCK-8 was used to determine the cell damage in each group after the use of MST4 inhibitor(HP),and western blot was used to determine the change of MST4 expression in each group.After mi R-486-5p was up-regulated or down-regulated using mimic and inhibitor transfected HT-22 cells,the expression level of MST4 was detected by western blot,and the cell damage in each group was measured by CCK-8.Results: The results of bioinformatics analysis suggested that mi R-486-5p was differentially expressed after acute hemorrhage and was correlated with MST4.The 3 ’-UTR of MST4 has a conserved mi R-486-5p binding site,and the results of dual luciferase reporter gene detection showed that: mi R-486-5p mimic was co-transfected with wild-type MST4 double luciferase plasmids and its fluorescence values decreased(P<0.05).When co-transfected with mutant MST4 double luciferase plasmid,the fluorescence value did not change significantly(P>0.05).The serum mi R-486-5p expression in patients with acute cerebral hemorrhage was significantly lower than that in healthy people(P<0.05).Serum mi R-486-5p levels were predictive of acute intracerebral hemorrhage(AUC=0.8939,95% confidence interval:0.8148-0.9729,sensitivity: 0.9333,specificity: 0.7667,Yoden index 0.7).Serum mi R-486-5p level was negatively correlated with hematoma volume in patients with acute cerebral hemorrhage(r =-0.6513,p<0.0001).In HT-22 cells,the expression of mi R-486-5p was significantly decreased and the expression of MST4 was significantly increased after Hemin-induced bleeding injury(P<0.05).After the use of MST4 inhibitor,the expression of MST4 was decreased(P<0.05)and the degree of cell damage was reduced(P<0.05).After downregulating mi R-486-5p,the expression of MST4 was increased(P<0.05)and the degree of cell damage was increased(P<0.05).Conclusions: mi R-486-5p is lowly expressed after cerebral hemorrhage.mi R-486-5p expression level correlates with hematoma volume.MST4 expression is elevated after cerebral hemorrhage and MST4 inhibitors are cerebroprotective.mi R-486-5p can target binding and downregulate MST4 to regulate the pathological injury process and affect cell viability after cerebral hemorrhage.