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| 微RNA-219通过ERK 1/2通路改善炎症所致新生SD大鼠脑内少突胶质细胞成熟障碍 |
| 张绍卿1,2,杜敏1,3,刘兰1,4,徐颖1,2* |
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(1. 重庆医科大学附属儿童医院麻醉科, 重庆 400014;
2. 国家儿童健康与疾病临床医学研究中心, 重庆 400014;
3. 儿童发育疾病研究教育部重点实验室, 重庆 400014;
4. 儿科学重庆市重点实验室, 重庆 400014
*通信作者) |
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| 摘要: |
| 目的 建立新生SD大鼠炎症模型，初步探究微RNA-219（miR-219）促进少突胶质细胞成熟的机制。方法 将 60只SD大鼠随机分为对照组、脂多糖（LPS）组（LPS 0.15 mg/kg腹腔注射）、LPS＋miR-219 agomir组（LPS 0.15 mg/kg腹腔注射，miR-219 agomir 3 μL侧脑室注射）、miR-219 antagomir组（miR-219 antagomir 3 μL侧脑室注射）和LPS＋miR-219 agomir＋U0126组（LPS 0.15 mg/kg腹腔注射，miR-219 agomir 3 μL侧脑室注射，U0126 30 mg/kg腹腔注射）。于大鼠出生第7天和第14天处死后取脑组织，采用qPCR检测脑组织中miR-219及炎症因子IL-1β、TNF-α的mRNA表达水平，蛋白质印迹法检测少突胶质细胞成熟标志物髓鞘碱性磷脂蛋白（MBP）和ERK 1/2表达水平，免疫荧光检测大鼠胼胝体少突胶质细胞的数量。结果 与对照组相比，大鼠腹腔注射LPS后脑组织内IL-1β、TNF-α mRNA表达均升高（P＜0.01），miR-219表达减少（P＜0.01）。与LPS组相比，在用miR-219 agomir提升大鼠脑内miR-219的表达后MBP和ERK 1/2蛋白表达均增加（P＜0.01），胼胝体少突胶质细胞数量增加。与对照组相比，在用miR-219 antagomir降低大鼠脑内miR-219的表达后，MBP和ERK 1/2蛋白表达均减少（P＜0.01），胼胝体少突胶质细胞数量减少。用ERK 1/2通路抑制剂U0126处理后，miR-219对少突胶质细胞的促成熟作用受到了抑制（P＜0.01）。结论 在大鼠脑内miR-219对少突胶质细胞的促成熟作用是通过ERK 1/2通路发挥作用的。 |
| 关键词: 炎症 脑白质损伤 微RNA-219 细胞外信号调控的激酶 少突胶质细胞 |
| DOI：10.16781/j.CN31-2187/R.20220193 |
| 投稿时间：2022-03-06修订日期：2022-09-02 |
| 基金项目:重庆市科学技术委员会民生项目(cstc2018jscx-msybX0104). |
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| MicroRNA-219 promotes oligodendrocyte maturation through ERK 1/2 signaling pathway in inflammation model of neonatal SD rats |
| ZHANG Shaoqing1,2,DU Min1,3,LIU Lan1,4,XU Ying1,2* |
(1. Department of Anesthesiology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China;
2. National Clinical Research Center for Child Health and Disorders, Chongqing 400014, China;
3. Key Laboratory of Child Development and Disorders of Ministry of Education, Chongqing 400014, China;
4. Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China
*Corresponding author) |
| Abstract: |
| Objective To establish an inflammation model in neonatal SD rats and to explore the mechanism by which microRNA-219 (miR-219) promotes oligodendrocyte maturation. Methods Sixty SD rats were randomly assigned to control group, lipopolysaccharide ［LPS］ group (LPS 0.15 mg/kg, intraperitoneal injection), LPS＋miR-219 agomir group (LPS 0.15 mg/kg, intraperitoneal injection; miR-219 agomir 3 μL, intraventricular injection), miR-219 antagomir group (miR-219 antagomir 3 μL, intraventricular injection), or LPS＋miR-219 agomir＋U0126 group (LPS 0.15 mg/kg, intraperitoneal injection; miR-219 agomir 3 μL, intraventricular injection; U0126 30 mg/kg, intraperitoneal injection). The rats were sacrified on the 7th and 14th day of life and the brain tissue was harvested. The expression levels of miR-219 and inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) mRNA were detected by quantitative polymerase chain reaction. The expression levels of myelin basic protein (MBP) and extracellular signal-regulated kinase 1/2 (ERK 1/2) were detected by Western blotting. The number of oligodendrocytes in the corpus callosum was observed by immunofluorescence. Results Compared with the control group, the expression levels of IL-1β and TNF-α mRNA were significantly increased after intraperitoneal injection of LPS in rats (both P＜0.01), and the expression of miR-219 was significantly decreased (P＜0.01). Compared with the LPS group, the expression levels of MBP and ERK 1/2 were significantly increased after miR-219 agomir was used to enhance the expression of miR-219 in the brain of rats (both P＜0.01), with increased oligodendrocytes in the corpus callosum. Compared with the control group, the expression levels of MBP and ERK 1/2 were significantly decreased after miR-219 antagomir was used to reduce the expression of miR-219 in the brain of rats (both P＜0.01), with decreased oligodendrocytes in the corpus callosum. After treatment with the ERK 1/2 pathway inhibitor U0126, the maturation promoting effect of miR-219 on oligodendrocytes was significantly inhibited (P＜0.01). Conclusion miR-219 can promote the maturation of oligodendrocytes in rat brain through the ERK 1/2 pathway. |
| Key words: inflammation white matter injury microRNA-219 extracellular signal-regulated kinase oligodendrocytes |
