Medical Clinical Laboratory, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(4):1681-1687. doi: 10.26355/eurrev_201902_17130.
The aim was to investigate the role of microRNA-26b-5p in regulating mesenchymal stem cells (MSCs) differentiation to type II of alveolar epithelial cells (AECII) in the disease course of neonatal respiratory distress syndrome (NRDS).
MSCs were first derived from rat bone marrow. In vitro induction of MSCs differentiation to AECII was conducted by SAGM. The mRNA levels of microRNA-26b-5p, Wnt5a, and AECII-related genes (Occludin, KGF, CK18, SpA, SpB, and SpC) during the process of cell differentiation were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was conducted for detecting levels of inflammatory factors tumor necrosis factor-α (TNF-α), interferon-α (INF-α), and interleukin-1 (IL-1) in cell supernatant. Dual-luciferase reporter gene assay was then carried out to verify the regulatory effect of microRNA-26b-5p on Wnt5a. MicroRNA-26b-5p expression in serum samples of NRDS neonates and healthy neonates was detected by qRT-PCR as well.
MicroRNA-26b-5p was overexpressed in NRDS neonates than those of healthy neonates. Besides, microRNA-26b-5p was highly expressed in the process of MSCs differentiation to AECII. MicroRNA-26b-5p overexpression remarkably inhibited AECII differentiation and Wnt5a expression. Levels of TNF-α, INF-α, and IL-1 in cell supernatant during differentiation induction were elevated. The regulatory effects of microRNA-26b-5p on AECII differentiation, Wnt5a expression, and inflammatory response were reversed by Wnt5a overexpression.
MicroRNA-26b-5p inhibits MSCs differentiation to AECII via inhibiting Wnt5a expression through the Wnt pathway.
研究 microRNA-26b-5p 在调节骨髓间充质干细胞（MSCs）向Ⅱ型肺泡上皮细胞（AECII）分化中的作用，探讨其在新生儿呼吸窘迫综合征（NRDS）发病过程中的作用。
从大鼠骨髓中提取 MSCs，采用 SAGM 体外诱导 MSCs 向 AECII 分化，用实时荧光定量聚合酶链反应（qRT-PCR）检测细胞分化过程中 microRNA-26b-5p、Wnt5a 及 AECII 相关基因（Occludin、KGF、CK18、SpA、SpB、SpC）的 mRNA 水平，酶联免疫吸附试验（ELISA）检测细胞上清液中炎症因子肿瘤坏死因子-α（TNF-α）、干扰素-α（INF-α）和白细胞介素-1（IL-1）的水平，双荧光素酶报告基因实验验证 microRNA-26b-5p 对 Wnt5a 的调控作用，采用 qRT-PCR 检测 NRDS 新生儿与健康新生儿血清样本中 microRNA-26b-5p 的表达。
NRDS 新生儿血清中 microRNA-26b-5p 的表达高于健康新生儿，且在 MSCs 向 AECII 分化过程中表达升高。过表达 microRNA-26b-5p 显著抑制 AECII 分化及 Wnt5a 表达，诱导分化过程中细胞上清液中 TNF-α、INF-α、IL-1 水平升高，而过表达 Wnt5a 则逆转了 microRNA-26b-5p 对 AECII 分化、Wnt5a 表达及炎症反应的调控作用。
microRNA-26b-5p 通过 Wnt 通路抑制 Wnt5a 表达，抑制 MSCs 向 AECII 分化。
