微小RNA miR-24-3p通过靶向肿瘤坏死因子受体相关因子6介导脂多糖诱导的子宫内膜炎症反应的负调控。

MicroRNA miR-24-3p Mediates the Negative Regulation of Lipopolysaccharide-Induced Endometrial Inflammatory Response by Targeting TNF Receptor-Associated Factor 6 ().

作者信息

Oladejo Ayodele Olaolu, Li Yajuan, Imam Bereket Habte, Ma Xiaoyu, Shen Wenxiang, Wu Xiaohu, Jiang Wei, Yang Jie, Lv Yanan, Ding Xuezhi, Wang Shengyi, Yan Zuoting

机构信息

Key Laboratory of Veterinary Pharmaceutical Development of Ministry of Agriculture, Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Science, Lanzhou, 730050, People's Republic of China.

Department of Animal Health Technology, Oyo State College of Agriculture and Technology, Igboora, 201103, Nigeria.

出版信息

J Inflamm Res. 2022 Feb 6;15:807-825. doi: 10.2147/JIR.S347293. eCollection 2022.

PURPOSE

Endometritis is a female reproductive disease that affects the cattle industries development and microRNAs (miRNAs) play a pivotal role and critical regulators of the innate immune response in varieties of diseases. The present study intends to investigate the regulatory role of miR-24-3p in the innate immune response involved in endometritis and evaluate its therapeutic potential.

METHODS

Whole mice uteri and bovine endometrial epithelial cells (BEECs) were separately stimulated with LPS. The BEECs were also transfected with miR-24-3p mimic and negative control; si and siNC; pcDNA3.1 empty and pcDNA3.1(+) separately with LPS stimulation. The expression levels of miR-24-3p and were measured via quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. LPS-induced inflammatory response assessed by inflammatory cytokines secretion and expression via ELISA and qRT-PCR. Bioinformatics analysis and luciferase reporter assay validated the interaction between miR-24-3p and . The activation of the NF-ĸB/MAPK pathway and p65 phosphorylation was investigated by Western blot and immunofluorescence assay, respectively.

RESULTS

The expression of miR-24-3p was decreased, and was elevated with an increased level of pro-inflammatory cytokines in LPS-treated BEECs and mice uterus. The overexpression of miR-24-3p suppressed LPS-induced secretion of inflammatory cytokines (IL-1β, IL-6, IL-8 and TNF-α) and deactivation of NF-ĸB/MAPK pathways. The downregulation of inhibited LPS-induced inflammatory response in BEECs. is validated as a target of miR-24-3p, and miR-24-3p reversed the overexpression of cloned on inflammation response in BEECs.

CONCLUSION

Our findings demonstrate that the overexpression of miR-24-3p attenuates endometrial inflammation and the expression of pro-inflammatory mediators via suppressing . Therefore, modulating the pathogenesis of endometritis and possibly, a therapeutic potential against endometritis.

目的

子宫内膜炎是一种影响养牛业发展的雌性生殖疾病，微小RNA（miRNA）在多种疾病的固有免疫反应中起关键作用并作为关键调节因子。本研究旨在探讨miR-24-3p在子宫内膜炎相关固有免疫反应中的调节作用，并评估其治疗潜力。

方法

用脂多糖（LPS）分别刺激小鼠全子宫和牛子宫内膜上皮细胞（BEECs）。BEECs还用miR-24-3p模拟物和阴性对照；小干扰RNA（si）和小干扰RNA阴性对照（siNC）；空的pcDNA3.1和pcDNA3.1（+）分别进行转染并给予LPS刺激。分别通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测miR-24-3p和[此处原文缺失相关基因名称]的表达水平。通过酶联免疫吸附测定（ELISA）和qRT-PCR检测LPS诱导的炎症反应，评估炎症细胞因子的分泌和表达。生物信息学分析和荧光素酶报告基因检测验证了miR-24-3p与[此处原文缺失相关基因名称]之间的相互作用。分别通过蛋白质免疫印迹法和免疫荧光测定法研究核因子κB（NF-κB）/丝裂原活化蛋白激酶（MAPK）途径的激活和p65磷酸化。

结果

在LPS处理的BEECs和小鼠子宫中，miR-24-3p的表达降低，[此处原文缺失相关基因名称]升高，促炎细胞因子水平增加。miR-24-3p的过表达抑制了LPS诱导的炎症细胞因子（白细胞介素-1β、白细胞介素-6、白细胞介素-8和肿瘤坏死因子-α）的分泌以及NF-κB/MAPK途径的失活。[此处原文缺失相关基因名称]的下调抑制了BEECs中LPS诱导的炎症反应。[此处原文缺失相关基因名称]被验证为miR-24-3p的靶标，并且miR-24-3p逆转了克隆的[此处原文缺失相关基因名称]在BEECs中对炎症反应的过表达。