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miRNA-221 通过靶向 TNF-α 表达调控脊髓损伤诱导的炎症反应。

miRNA-221 Regulates Spinal Cord Injury-Induced Inflammatory Response through Targeting TNF- Expression.

机构信息

Department of Spine Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, 150001 Heilongjiang Province, China.

Department of Orthopaedic Surgery, General Hospital of General Administration of Agriculture and Reclamation, Harbin, Heilongjiang Province 150088, China.

出版信息

Biomed Res Int. 2021 Apr 7;2021:6687963. doi: 10.1155/2021/6687963. eCollection 2021.

DOI:10.1155/2021/6687963
PMID:33928162
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8049790/
Abstract

OBJECTIVES

To investigate the roles of miR-221 in spinal cord injury (SCI) as well as the underlying mechanism.

METHODS

A mouse model of SCI was generated and used to examine dynamic changes in grip strength of the mouse upper and lower limbs. The expression of miR-221 and tumor necrosis factor- (TNF-) was detected by RT-qPCR and Western blot. Levels of inflammation and oxidative stress in microglia cells of the injured mice overexpressing miR-221 were then measured by ELISA. Bioinformatics analysis and dual-luciferase reporter assay were conducted to identify the miR-221 target.

RESULTS

We successfully constructed SCI mouse model. The results of qRT-PCR showed that miR-221 was gradually upregulated in the spinal cord tissue of mice in the SCI group with the prolonged injury time. At the same time, the mRNA and protein of TNF- gradually decreased. We further confirmed through cell experiments that the inflammatory factors TNF- and IL-6, as well as iNOS and eROS, were upregulated in spinal cord microglia cells of SCI mice, and upregulation of miR-122 can inhibit their expression. Finally, the luciferase reporter experiment confirmed that miR-122 targeted TNF-.

CONCLUSIONS

We present evidence that miR-221 promotes functional recovery of the injured spinal cord through targeting TNF-, while alleviating inflammatory response and oxidative stress.

摘要

目的

研究 miR-221 在脊髓损伤 (SCI) 中的作用及其机制。

方法

构建 SCI 小鼠模型，检测小鼠上下肢抓力的动态变化。采用 RT-qPCR 和 Western blot 检测 miR-221 和肿瘤坏死因子- (TNF-) 的表达。通过 ELISA 检测过表达 miR-221 的损伤小鼠小胶质细胞中的炎症和氧化应激水平。通过生物信息学分析和双荧光素酶报告基因实验鉴定 miR-221 的靶基因。

结果

我们成功构建了 SCI 小鼠模型。qRT-PCR 结果显示，随着损伤时间的延长，SCI 组小鼠脊髓组织中 miR-221 逐渐上调，同时 TNF- 的 mRNA 和蛋白逐渐下降。我们通过细胞实验进一步证实，SCI 小鼠脊髓小胶质细胞中的炎症因子 TNF- 和 IL-6 以及 iNOS 和 eROS 表达上调，而过表达 miR-122 可以抑制其表达。最后，荧光素酶报告实验证实 miR-122 靶向 TNF-。

结论

我们的研究结果表明，miR-221 通过靶向 TNF- 促进损伤脊髓的功能恢复，同时减轻炎症反应和氧化应激。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/872df03677e2/BMRI2021-6687963.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/f9c9aa111d65/BMRI2021-6687963.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/117154079c6c/BMRI2021-6687963.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/84d8c883cf86/BMRI2021-6687963.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/872df03677e2/BMRI2021-6687963.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/f9c9aa111d65/BMRI2021-6687963.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/117154079c6c/BMRI2021-6687963.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/84d8c883cf86/BMRI2021-6687963.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/553f/8049790/872df03677e2/BMRI2021-6687963.004.jpg

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