Effects of Exercise Preconditioning on the Inflammatory Response of Rats after Ischemic Stroke through Exosome-mediated miR-146a
10.3969/j.issn.1000-6710.2023.11.007
- VernacularTitle:运动预处理通过外泌体介导miR-146a对大鼠缺血性脑卒中炎症反应的影响
- Author:
Lu WANG
1
;
Dong YAN
;
Ruijie XIA
;
Caiyun JIANG
;
Jingjing CHU
;
Jinmei ZHANG
;
Wenqian DENG
Author Information
1. 成都体育学院(成都 610041)
- Keywords:
exercise preconditioning;
ischemic stroke;
miR-146a;
exosomes
- From:
Chinese Journal of Sports Medicine
2023;42(11):889-898
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of exercise preconditioning on inflammatory response in ischemic stroke brain tissue which mediated by miR-146a in extracellular vesicles in rats with middle cerebral artery oc-clusion(MCAO),and its mechanism.Methods Sixty 6-week-old male Sprague-Dawley rats were randomly divid-ed into a non-exercise group and an exercise group.The non-exercise group was further divided into a sham-operation control group(C,n=15)and an MCAO model group(M,n=15),while the exercise group was further di-vided into an exercise only group(E,n=15)and an exercise plus MCAO model group(EM,n=15).Rats in the E and EM groups underwent 8 weeks of treadmill exercise,6 days per week,30 minutes per day.Then rats in the M and EM groups received MCAO to induce ischemic stroke,while the C and E groups underwent a sham surgery.Twenty-four hours after the surgery,neurobehavioral tests were performed.Plasma was collected to ex-tract extracellular vesicles,and brain tissue was collected to measure the volume of cerebral infarction by using the triphenyl tetrazolium chloride(TTC)staining.Moreover,the Nissl staining was conducted to observe neuronal and Nissl body.Mean while,the content of miR-146a in plasma extracellular vesicles and brain tissue was mea-sured by using the quantitative polymerase chain reaction(qPCR),and the expression of TNF receptor associat-ed factor 6(TRAF6),nuclear factor kappa-B(NF-κB)and tumor necrosis factor-α(TNF-α)in brain tissues were determined using Western blotting.The targeting relationship between miR-146a and TRAF6 was detected by using the dual luciferase reporter gene assay.Results(1)The neurological behavioral scores of the EM and M groups were higher than those of the C group(P<0.01 and P<0.01),with that of the EM group lower than the M group(P<0.01).(2)TTC staining showed that the infarct volume of the EM and M groups was larger than that of the other two groups(P<0.01 and P<0.01),with that of the EM group smaller than the M group(P<0.01).(3)Nissl staining results showed that the neuronal arrangement was loose,the number of neurons re-duced,and the Nissl bodies were lightly stained and decreased in the M group compared with the C and E groups.Moreover,compared with the M group,the number of neurons and Nissl bodies increased in the EM group.(4)The qPCR analysis showed that the expression of miR-146a in the plasma-derived exosomes and brain tissues of the EM and M groups decreased compared with the C group(P<0.05 and P<0.01),with that of the EM group higher than the M group(P<0.05).(5)According to Western blotting,compared with the C group,the expression levels of TRAF6,NF-κB,and TNF-α proteins increased significantly(P<0.05 and P<0.01),with that of group EM signfiicantly lower than group M(P<0.05 and P<0.05).(6)Dual-luciferase report-er gene assay showed that miR-146a had a specific binding site with TRAF6.Conclusion Eight weeks of exer-cise preconditioning reduces the infarct area and the extent of brain damage,which may be mediated by miR-146a via exosomes,increasing the expression of miR-146a in brain tissue,targeting TRAF6,negatively regulat-ing TRAF6/NF-κB,and reducing the expression of TNF-α,thus alleviating the inflammatory response in brain tissue and exerting a protective effect on ischemic brain injury.
