The 958th PLA Hospital, Chongqing, China.
Eur Rev Med Pharmacol Sci. 2018 Nov;22(22):7849-7857. doi: 10.26355/eurrev_201811_16410.
To explore whether miR-216a-3p could promote differentiation of bone marrow mesenchymal stem cells (BMMSCs) into type II alveolar epithelial cells (ACE II) via Wnt/β-catenin pathway, thereby alleviating neonatal respiratory distress syndrome (NRDS).
BMMSCs were directionally differentiated into ACE II cells. Expressions of ACE II cell-specific transcription factors Occludin, KGF, CK18, SpA, SpB, and SpC were detected at the different time points after cell differentiation. Enzyme-linked immunosorbent assay (ELISA) was applied to detect inflammatory factors in the culture medium, including interleukin-1 (IL-1), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α), and Interferon-α (INF-α). After overexpression or knockdown of miR-216a-3p in BMMSCs, expressions of ACE II cell-specific transcription factors and inflammatory factors were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. Rescue experiments were carried out after DKK-1 treatment in BMMSCs.
Expressions of ACE II cell-specific transcription factors Occludin, KGF, CK18, SpA, SpB, and SpC were elevated with the prolongation of cell differentiation. Overexpression of miR-216a-3p elevated levels of pro-inflammatory factors (IL-1, TNF-α, and INF-α) and reduced anti-inflammatory factor (IL-10). Expressions of ACE II cell-specific transcription factors Occludin, KGF, CK18, SpA, SpB, and SpC were remarkably increased at 7 d and 14 d compared to those detected at 1 d. Overexpression of miR-216a-3p in BMMSCs downregulated Wnt3a expression. The regulatory effect of miR-216a-3p on BMMSCs differentiation was partially reversed by DKK-1 treatment.
Knockdown of miR-216a-3p induces differentiation of BMMSCs into ACE II cells through Wnt/β-catenin pathway, thereby alleviating NRDS.
探讨 miR-216a-3p 是否可通过 Wnt/β-catenin 通路促进骨髓间充质干细胞（BMMSCs）向 II 型肺泡上皮细胞（ACE II）分化，从而缓解新生儿呼吸窘迫综合征（NRDS）。
定向诱导 BMMSCs 分化为 ACE II 细胞。在细胞分化后的不同时间点检测 ACE II 细胞特异性转录因子 Occludin、KGF、CK18、SpA、SpB 和 SpC 的表达。采用酶联免疫吸附试验（ELISA）检测培养基中的炎症因子，包括白细胞介素 1（IL-1）、白细胞介素 10（IL-10）、肿瘤坏死因子-α（TNF-α）和干扰素-α（INF-α）。在 BMMSCs 中转染 miR-216a-3p 过表达或敲低后，通过定量实时聚合酶链反应（qRT-PCR）和 Western blot 检测 ACE II 细胞特异性转录因子和炎症因子的表达。在 BMMSCs 中转染 DKK-1 后进行挽救实验。
随着细胞分化时间的延长，ACE II 细胞特异性转录因子 Occludin、KGF、CK18、SpA、SpB 和 SpC 的表达逐渐升高。miR-216a-3p 过表达增加了促炎因子（IL-1、TNF-α 和 INF-α）的水平，降低了抗炎因子（IL-10）的水平。与 1 d 相比，在 7 d 和 14 d 时 ACE II 细胞特异性转录因子 Occludin、KGF、CK18、SpA、SpB 和 SpC 的表达明显升高。miR-216a-3p 在 BMMSCs 中的过表达下调了 Wnt3a 的表达。DKK-1 处理部分逆转了 miR-216a-3p 对 BMMSCs 分化的调节作用。
miR-216a-3p 敲低通过 Wnt/β-catenin 通路诱导 BMMSCs 向 ACE II 细胞分化，从而缓解 NRDS。
