Protective effect of microRNA-134-3p on multiple sclerosis through inhibiting PRSS57 and promotion of CD34+ cell proliferation in rats.,Journal of Cellular Biochemistry

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Protective effect of microRNA-134-3p on multiple sclerosis through inhibiting PRSS57 and promotion of CD34+ cell proliferation in rats.
Journal of Cellular Biochemistry ( IF 3.0 ) Pub Date : 2020-07-03 , DOI: 10.1002/jcb.29643
Qihan Song _{1,

2} , Fengli Zhao ₂ , Jingfan Yao ₃ , Hailin Dai ₂ , Lei Hu ₂ , Shun Yu ₁

Affiliation

MicroRNAs (miRs) have been extensively studied for their involvement in multiple sclerosis (MS). We investigated the involvement of miR‐134‐3p on MS. The MS rat model was established, and positive expression of interleukin‐17 (IL‐17) was detected using the immunohistochemical method while the expression of miR‐134‐3p and serine protease 57 (PRSS57) was determined by means of reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) and Western blot analysis. Second, the miR‐134‐3p overexpression or short hairpin RNA against PRSS57 was introduced into the CD34⁺ cells to investigate the levels of proliferation and apoptosis‐related genes by RT‐qPCR and Western blot analysis. In addition, analysis of the targeting relations of miR‐134‐3p and PRSS57 was conducted using online software and dual‐luciferase reporter gene assay. Furthermore, neuronal functions, inflammatory response, proliferation, and apoptosis of CD34⁺ cells were assayed by flow cytometry, enzyme‐linked immunosorbent assay, and methyl thiazolyl tetrazolium. IL‐17 and PRSS57 expression increased while miR‐134‐3p expression decreased in the spinal cord from MS rats. miR‐134‐3p could target PRSS57. miR‐134‐3p overexpression or PRSS57 silencing enhanced mitochondrial activity of neurons, mitochondrial membrane potential content, CD34⁺ cell proliferation, while decreasing Cyt C content, inflammatory response, and cell apoptosis. Collectively, overexpression of miR‐134‐3p promotes CD34⁺ cell proliferation via inhibition of PRSS57 in MS, which may serve as a promising target for MS intervention.

中文翻译：

microRNA-134-3p通过抑制PRSS57和促进CD34+细胞增殖对大鼠多发性硬化的保护作用。

MicroRNAs (miRs) 因其参与多发性硬化症 (MS) 而被广泛研究。我们研究了 miR-134-3p 对 MS 的影响。建立MS大鼠模型，免疫组化法检测白细胞介素17（IL-17）的阳性表达，逆转录-定量法检测miR-134-3p和丝氨酸蛋白酶57（PRSS57）的表达。聚合酶链反应 (RT-qPCR) 和蛋白质印迹分析。其次，将针对 PRSS57 的 miR-134-3p 过表达或短发夹 RNA 引入 CD34 ⁺通过 RT-qPCR 和蛋白质印迹分析研究增殖和凋亡相关基因的水平。此外，使用在线软件和双荧光素酶报告基因检测分析了 miR-134-3p 和 PRSS57 的靶向关系。此外，CD34 ⁺细胞的神经元功能、炎症反应、增殖和凋亡通过流式细胞术、酶联免疫吸附试验和甲基噻唑基四唑进行了测定。在 MS 大鼠的脊髓中，IL-17 和 PRSS57 表达增加，而 miR-134-3p 表达减少。miR-134-3p 可以靶向 PRSS57。miR-134-3p 过表达或 PRSS57 沉默增强神经元线粒体活性、线粒体膜电位含量、CD34 ⁺细胞增殖，同时降低 Cyt C 含量、炎症反应和细胞凋亡。总的来说，miR-134-3p 的过表达通过抑制 MS 中的 PRSS57 促进 CD34 ⁺细胞增殖，这可能是 MS 干预的一个有希望的靶点。

更新日期：2020-07-03

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