Singh Naresh, Nagar Ekta, Gautam Anshu, Kapoor Himanshi, Arora Naveen
CSIR-Institute of Genomics and Integrative Biology, Delhi 110007, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad 201002, India.
CSIR-Institute of Genomics and Integrative Biology, Delhi 110007, India.
Sci Total Environ. 2023 Dec 1;902:166063. doi: 10.1016/j.scitotenv.2023.166063. Epub 2023 Aug 5.
Diesel exhaust (DE) exposure contributes to the progression of chronic respiratory diseases and is associated with dysregulation of microRNA expression. The present study aims to investigate the involvement of miRNAs and target genes in DE-induced lung fibrosis.
C57BL/6 mice were divided into three groups. Group 1 mice were exposed to filtered air (Control). Group 2 mice were exposed to DE for 30 min per day, 5 days per week, for 8 weeks (DE). Group 3 mice received DE exposure along with resveratrol on alternate days for the last 2 weeks (DE + RES). Mice were sacrificed to isolate RNA from lung tissue for miRNA microarray profiling. Bronchoalveolar lavage fluid and lung tissues were collected for cell count and biochemical analysis.
DE exposure resulted in differential expression of 28 miRNAs with fold change >2 (p < 0.05). The upregulated miR-212-3p was selected for further analysis. Consensus analysis revealed enrichment of SIRT1 in the FoxO pathway, along with a co-annotation of reduced body weight (p < 0.05). A549 cells transfected with a miR-212-3p inhibitor showed a dose-dependent increase in SIRT1 expression, indicating SIRT1 as a direct target. Treatment with resveratrol restored SIRT1 and miR-212-3p expression and led to a reduction in inflammatory cytokines (p < 0.05). The modulation of SIRT1 correlated negatively with macrophage infiltration, confirming its role in regulating cellular infiltration and lung inflammation. Fibronectin, alpha-SMA, and collagen levels were significantly decreased in DE + RES compared to DE group suggesting modulation of cellular functions and resolution of lung fibrosis. Furthermore, a significant decrease in FoxO3a and TGF-β gene expressions was observed upon resveratrol administration thereby downregulating pro-fibrotic pathway.
The present study demonstrates resveratrol treatment stabilizes SIRT1 gene expression by attenuating miR-212-3p in DE-exposed mice, leading to downregulation of TGF-β and FoxO3a expressions. The study highlights the therapeutic role of resveratrol in the treatment of DE-induced pulmonary fibrosis.
接触柴油机尾气（DE）会促使慢性呼吸道疾病进展，且与微小RNA（miRNA）表达失调有关。本研究旨在探究miRNA及其靶基因在DE诱导的肺纤维化中的作用。
将C57BL/6小鼠分为三组。第1组小鼠暴露于过滤空气（对照组）。第2组小鼠每周5天，每天暴露于DE 30分钟，共8周（DE组）。第3组小鼠在最后2周隔天接受DE暴露并同时给予白藜芦醇（DE + RES组）。处死小鼠以从肺组织中分离RNA用于miRNA微阵列分析。收集支气管肺泡灌洗液和肺组织进行细胞计数和生化分析。
DE暴露导致28种miRNA表达差异，变化倍数>2（p < 0.05）。上调的miR-212-3p被选作进一步分析。一致性分析显示在FoxO通路中沉默信息调节因子1（SIRT1）富集，同时伴有体重减轻的共同注释（p < 0.05）。用miR-212-3p抑制剂转染的A549细胞显示SIRT1表达呈剂量依赖性增加，表明SIRT1是直接靶标。白藜芦醇处理可恢复SIRT1和miR-212-3p表达，并导致炎性细胞因子减少（p < 0.05）。SIRT1的调节与巨噬细胞浸润呈负相关，证实其在调节细胞浸润和肺部炎症中的作用。与DE组相比，DE + RES组中纤连蛋白、α-平滑肌肌动蛋白和胶原蛋白水平显著降低，提示细胞功能得到调节且肺纤维化得到缓解。此外，给予白藜芦醇后观察到FoxO3a和转化生长因子-β（TGF-β）基因表达显著降低，从而下调促纤维化通路。
本研究表明，白藜芦醇治疗通过减弱DE暴露小鼠中的miR-212-3p来稳定SIRT1基因表达，导致TGF-β和FoxO3a表达下调。该研究突出了白藜芦醇在治疗DE诱导的肺纤维化中的治疗作用。
