Micro RNA-193b-3p inhibits neuro-inflammatory response in neonatal sepsis rats by targeting retinoic acid receptor-related orphan receptor α
10.3760/cma.j.cn115354-20210603-00355
- VernacularTitle:miR-193b-3p通过靶向RORα抑制新生脓毒症大鼠的神经炎症反应
- Author:
Ziyan ZHAO
1
;
Jianwei SUN
;
Dandan WANG
;
Lei ZHANG
;
Jiajie ZHANG
Author Information
1. 河南省人民医院儿科，郑州 450003
- Keywords:
Micro RNA-193b-3p;
Neuroinflammatory response;
Sepsis;
Neonatal rat;
Neurocyte
- From:
Chinese Journal of Neuromedicine
2021;20(11):1092-1100
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the micro RNA (miR)-193b-3p expression in hippocampus tissues of neonatal sepsis rats, as well as its role in neuroinflammatory response and possible mechanisms.Methods:Twenty-four 2-d-old SD rats were randomly divided into control group, lipopolysaccharide (LPS) group, negative control group, and miR-193b-3p group ( n=6); except for the negative control group, the rats in the other 3 groups were intraperitoneally injected with LPS to establish sepsis models; in miR-193b-3p group and NC group, 5 μL miR-193b-3p mimics/controls (20 nmol/L)were injected into the lateral ventricle 3 d before LPS injection. In vitro, PC12 cells were divided into control group, LPS group, miR-193b-3p group, and LPS+miR-193b-3p group; miR-193b-3p mimics were transfected into cells of miR-193b-3p group and LPS+miR-193b-3p group; cells in the LPS group and LPS+miR-193b-3p group were exposed to 100 ng/mL LPS. The miR-193b-3p downstream target gene was analyzed by gene microarray and dual luciferase reporter. Neurobehavioral scale used to assess the neurological function at specific time points in each group. The mRNA expression levels of miR-193b-3p and cytokines (interleukin [IL]-1β, IL-6, tumor necrosis factor [TNF]-α) in the brain tissues or PC12 cells were analyzed by real-time fluorescence quantification PCR (RT-qPCR). The protein expression levels of retinoic acid-related orphan receptor α (RORα), neuronal nuclear antigen (NeuN), ionized calcium binding adaptor molecule-1 (IBA-1) and glial fibrillary acidic protein (GFAP) were detected by double-labelling immunofluorescence. The protein expression levels of RORα in PC12 cells were detected by immunofluorescence staining. Results:(1) Gene microarray and dual luciferase reporter analysis confirmed that RORα was the target gene of miR-193b-3p. (2) As compared with those in rats of the negative control group, the neurobehavioral scores in rats of the LPS group were significantly decreased since 6 h of LPS injection and reached to the lowest at 24 h after LPS ( P<0.05). As compared with those in the negative control group, the IL-1β, IL-6, and TNF-α mRNA expression levels in the hippocampus of LPS group were significantly increased, while the miR-193b-3p expression was significantly decreased ( P<0.05). (3) As compared with those in negative control group (3.23±0.92), the neurobehavioral scores in rats of the miR-193b-3p group (7.51±0.84) 48 h after LPS injection were significantly higher ( P<0.05). As compared with those in the negative control group, the IL-1β, IL-6 and TNF-α mRNA expression levels in the hippocampus of the miR-193b-3p group were significantly deceased, while the miR-193b-3p expression was significantly higher ( P<0.05). (4) As compared with that in negative control group, the number of NeuN(+)RORα(+) cells in the hippocampus of the LPS group was significantly reduced ( P<0.05); as compared with negative control group, miR-193b-3p group had significantly increased number of NeuN(+)RORα(+) cells in the hippocampus ( P<0.05). (5) As compared with the control group, the LPS group had significantly decreased RORα expression, and significantly increased TNF-α, IL-1β and IL-6 mRNA expression in PC12 cells ( P<0.05). As compared with those in the LPS group, the RORα expression was significantly increased, and the TNF-α, IL-1β and IL-6 mRNA expression levels in PC12 cells were significantly decreased in LPS+miR-193b-3p group ( P<0.05). Conclusion:The miR-193b-3p inhibits the neuroinflammatory response in neonatal sepsis rats by regulating its target molecule RORα mRNA expression in hippocampal neurocyte.
