MiR-30a targeting Keap1 in relieving cerebral ischemia-reperfusion injury in rats
10.3760/cma.j.cn431274-20190415-00437
- VernacularTitle:MiR-30a靶向Keap1缓解大鼠脑缺血再灌注损伤的机制研究
- Author:
Yakun WU
;
Jia YAO
;
Kai FENG
- From:
Journal of Chinese Physician
2021;23(1):63-67
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the function and mechanism of miR-30a in rat cerebral ischemia-reperfusion (I/R) injury.Methods:The middle cerebral artery occlusion (MCAO) model was established by intravascular suture method, and the expression of miR-30a in brain tissue was detected by real-time quantitative polymerase chain reaction (qRT-PCR). After intracerebroventricular injection of miR-30a lentivirus, the infarct area was detected by 2, 3, 5-triphenyltetrazole chloride (TTC) staining, the neurological deficit was detected by Bederson method, and the concentration of neurotrophin-3 (3-NT) and nitric oxide (NO) in brain tissue was detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of kelch like ECH associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf-2) and hemeoxygenase-1 (HO-1) in brain tissue were detected by Western blot. Double luciferase reporter assay was used to detect the targeting relationship between miR-30a and Keap1.Results:Compared with sham operation group, the expression of miR-30a was down-regulated in a time-dependent manner after I/R. The overexpression of miR-30a can reduce the area of cerebral infarction tissue at the pathological level, the degree of neurological impairment at the functional level, the 3-NT, NO and Keap1 at the molecular level, and enhance the expression of Nrf2 and HO-1. The dual luciferase reporter assay also showed that miR-30a could bind to Keap1 mRNA.Conclusions:The expression of miR-30a was down-regulated in MCAO rat brain tissue, and miR-30a could attenuate cerebral I/R injury in rats by targeting Keap1.
