| Objectives To explore the biological effects of micro RNA-124 on cerebral ischemiareperfusion injury in rats and its mechanism of action on cell pyroptosis,so as to provide a laboratory basis for clinical treatment of cerebral ischemia-reperfusion injury in the future.Methods Eighty eight-week-old male SD rats were randomly divided into normal control group,sham-operated group,ischemia-reperfusion group(divided into 3,6,12,24 hours after reperfusion),solvent control group(NC group),micro RNA-124 agonist group(agomicro RNA-124 group),micro RNA-124 inhibitor group(antago-micro RNA-124 group),STAT3 inhibitor group(S31-201 group),with 8 rats in each group.Models of middle cerebral artery occlusion in rats were established by Zea-Longa thread embolization.Hematoxylin-eosin.HE,TTC staining and neurological function scoring were used to evaluate the degree of brain injury at cellular,tissue and behavioral levels.The relative expression of micro RNA-124 in brain tissue was analyzed by q RT-PCR,and the downstream target gene of micro RNA-124(STAT3)was validated by luciferase reporter gene technology and Western blot technology.Immunohistochemical and Western blot techniques were used to further analyze the expression and distribution of caspase-1,Gasdermin D and pro-inflammatory factors IL-1beta and IL-18 in brain tissue.Results 1 Compared with the normal control group,there was no significant difference in neurological function score,micro RNA-124 and expression of cell pyroptosis-related protein in sham-operated group.The neurological function score,micro RNA-124 and expression of cell pyroptosis-related protein in model group increased significantly at different time points.The expression of micro RNA-124 peaked at 12 hours after reperfusion,decreased at 24 hours,and the expression of pyroptosis-related proteins showed an increasing trend within 24 hours(P < 0.05).2 Compared with the negative control group,the area of cerebral infarction(17.37±1.47 vs 38.30±3.44,P<0.05)and neurological function score(1.60 ± 0.54 vs 2.40 ± 0.55,P<0.05)in the high expression micro RNA-124 group were significantly decreased,and the number of positive cells expressing pyroptosis-related protein was significantly reduced by immunohistochemistry,while the above indexes in the micro RNA-124 inhibitor group were significantly increased(P < 0.05).3 In luciferase reporter gene technology,compared with the negative control group,the luciferase activity in the micro RNA-124 + wild type STAT3 plasmid group decreased significantly(0.50±0.02 vs 1.00±0.05,P<0.05),while the luciferase activity in the micro RNA-124 + mutant STAT3 plasmid group did not change significantly(0.95±0.03 vs 1.00±0.04,P>0.05).Western blot results showed that compared with the control group,the high expression of micro RNA-124 group STAT3 and p-STAT3 decreased significantly(0.27±0.02 vs 0.47±0.02;0.38±0.02 vs 0.46±0.03,P<0.05),the expression of pyroptosis-related protein in S31-201 group decreased significantly,Caspase-1p20(0.43±0.02 vs 0.80±0.03,P<0.05),Gasdermin D(0.34±0.03 vs 0.75±0.03,P<0.05),Activated-IL-1?(0.30±0.03 vs 0.71±0.03,P<0.05),Activated-IL-18(0.27±0.02 vs0.67±0.03,P<0.05).Conclusions MicroRNA-124 can play a neuroprotective role by negatively regulating STAT3 to reduce the occurrence of cell pyroptosis in cerebral ischemia-reperfusion injury,which provides a new target for micro RNA-124 in clinical treatment of cerebral ischemiareperfusion injury.Figure 31;Table 14;Reference 140... |
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