School of Anesthesiology, Xuzhou Medical University, Xuzhou, Jiangsu, China.
Eur Rev Med Pharmacol Sci. 2020 Aug;24(16):8483-8492. doi: 10.26355/eurrev_202008_22661.
Pulmonary fibrosis (PF) is a chronic lung disease with complex pathogenesis and poor prognosis. Studies had demonstrated that long non-coding RNAs (lncRNAs) play an important role in the development of fibrosis. We explored the roles of NEAT1 in PF progression in this study.
PF tissues and TGF-β1-induced cells were analyzed for the function of NEAT1 in PF progression. qRT-PCR or Western blot was applied to detect NEAT1, miR‑9-5p or protein expressions. PF mice model assay was used to detect the effects of NEAT1 on PF in vivo. Luciferase reporter assay was applied to confirm target relationship between NEAT1 and miR‑9-5p. Correlation of NEAT1 and miR-9-5p was analyzed by Spearman's method.
We observed that NEAT1 was significantly upregulated while miR-9-5p was downregulated in PF tissues and TGF-β1-induced cells. A negative correlation was exhibited of NEAT1 and miR-9-5p expression in PF tissues. Protein level of p-Smad2 was increased in TGF-β1 induced cells. Furthermore, NEAT1 knockdown increased E-cadherin expression, while decreased N-cadherin, Vimentin, Collagen I, Collagen III and α-smooth muscle actin (α-SMA) expressions in TGF-β1-induced cells. Moreover, NEAT1 could directly target miR-9-5p to regulate the PF induced by TGF-β1. The miR-9-5p overexpression inhibited TGF-β1 and p-Smad2 expression, while NEAT1 overexpression attenuated this effect. In addition, NEAT1 inhibition enhanced E-cadherin expression, and reduced TGF-β1, p-Smad2, N-cadherin, Collagen I, Collagen III, α-SMA and Vimentin expression after BLM treatment.
Taken together, our findings showed that NEAT1 knockdown attenuated PF via the regulatory of miR-9-5p and TGF-β signaling to repress EMT and might provide new therapeutic targets for PF patients.
肺纤维化（PF）是一种具有复杂发病机制和预后不良的慢性肺部疾病。研究表明，长链非编码 RNA（lncRNA）在纤维化的发展中起重要作用。本研究探讨了 NEAT1 在 PF 进展中的作用。
分析 PF 组织和 TGF-β1 诱导的细胞中 NEAT1 在 PF 进展中的作用。应用 qRT-PCR 或 Western blot 检测 NEAT1、miR-9-5p 或蛋白表达。应用 PF 小鼠模型实验检测 NEAT1 对体内 PF 的影响。应用荧光素酶报告基因实验证实 NEAT1 与 miR-9-5p 之间的靶关系。应用 Spearman 法分析 NEAT1 与 miR-9-5p 的相关性。
我们观察到，在 PF 组织和 TGF-β1 诱导的细胞中，NEAT1 显著上调，而 miR-9-5p 下调。PF 组织中 NEAT1 和 miR-9-5p 的表达呈负相关。TGF-β1 诱导的细胞中 p-Smad2 蛋白水平升高。此外，NEAT1 敲低可增加 TGF-β1 诱导的细胞中 E-钙黏蛋白的表达，而降低 N-钙黏蛋白、波形蛋白、I 型胶原、III 型胶原和α-平滑肌肌动蛋白（α-SMA）的表达。此外，NEAT1 可直接靶向 miR-9-5p 调节 TGF-β1 诱导的 PF。miR-9-5p 过表达抑制 TGF-β1 和 p-Smad2 的表达，而 NEAT1 过表达可减弱这种作用。此外，NEAT1 抑制可增强 BLM 处理后 E-钙黏蛋白的表达，降低 TGF-β1、p-Smad2、N-钙黏蛋白、I 型胶原、III 型胶原、α-SMA 和波形蛋白的表达。
综上所述，我们的研究结果表明，NEAT1 敲低通过调节 miR-9-5p 和 TGF-β 信号通路抑制 EMT 来减轻 PF，可能为 PF 患者提供新的治疗靶点。
