2. Academic Validation
  3. Silencing XIST mitigated lipopolysaccharide (LPS)-induced inflammatory injury in human lung fibroblast WI-38 cells through modulating miR-30b-5p/CCL16 axis and TLR4/NF-κB signaling pathway

Silencing XIST mitigated lipopolysaccharide (LPS)-induced inflammatory injury in human lung fibroblast WI-38 cells through modulating miR-30b-5p/CCL16 axis and TLR4/NF-κB signaling pathway

  • Open Life Sci. 2021 Feb 6;16(1):108-127. doi: 10.1515/biol-2021-0005.
Jiahui Xu 1 2,  Honggui Li 3,  Ying Lv 2,  Chang Zhang 4,  Yiting Chen 5,  Dezhao Yu 1
Affiliations

Affiliations

  • 1 Department of Pediatrics, Guangdong Second Hospital of Traditional Chinese Medicine, No. 60 Hengfu Road, 510000, Guangzhou, Guangdong, China.
  • 2 Department of Pediatrics, The Fifth Clinical College of Guangzhou University of Traditional Chinese Medicine, 510000, Guangzhou, Guangdong, China.
  • 3 Department of Pediatrics, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, 510000, Guangzhou, Guangdong, China.
  • 4 Department of Internal Medicine, Guangdong Second Hospital of Traditional Chinese Medicine, 510000, Guangzhou, Guangdong, China.
  • 5 Department of Pediatrics, The Affiliated Zhujiang Hospital of Southern Medical University, 510000, Guangzhou, Guangdong, China.
PMID: 33817304 DOI: 10.1515/biol-2021-0005
Abstract

Background: Emerging evidence shows that long noncoding RNA (lncRNA) has been a novel insight in various diseases, including pneumonia. Even though lncRNA X-inactive-specific transcript (XIST) is well studied, its role in pneumonia remains to be largely unrevealed.

Methods: Expression of XIST, miRNA-30b-5p (miR-30b-5p), and CC chemokine ligand 16 (CCL16) was detected using Reverse Transcriptase quantitative polymerase chain reaction and western blotting; their interaction was confirmed by dual-luciferase reporter assay. Apoptosis, inflammation, and Toll-like Receptor 4 (TLR4)/NF-κB signaling pathway were measured using methyl thiazolyl tetrazolium assay, flow cytometry, western blotting, and enzyme-linked immunosorbent assay.

Results: Lipopolysaccharide (LPS) stimulation decreased cell viability and B cell lymphoma (Bcl)-2 expression, and increased cell Apoptosis rate and expression of Bcl-2-associated X protein (Bax), cleaved-caspase-3, interleukin (IL)-6, IL-1β, and tumor necrosis factor α (TNF-α) in WI-38 cells. Expression of XIST and CCL16 was upregulated in the serum of patients with pneumonia and LPS-induced WI-38 cells, respectively; silencing XIST and CCL16 could suppress LPS-induced Apoptosis and inflammation in WI-38 cells, and this protection was abolished by miR-30b-5p downregulation. Moreover, XIST and CCL16 could physically bind to miR-30b-5p, and XIST regulated CCL16 expression via sponging miR-30b-5p. TLR4 and phosphorylated P65 (p-P65) and p-IκB-α were highly induced by LPS treatment, and this upregulation was diminished by blocking XIST, accompanied with CCL16 downregulation and miR-30b-5p upregulation.

Conclusions: Silencing XIST could alleviate LPS-induced inflammatory injury in human lung fibroblast WI-38 cells through modulating miR-30b-5p/CCL16 axis and inhibiting TLR4/NF-κB signaling pathway.

Keywords

CCL16; LPS-induced WI-38 cells; TLR4/NF-κB; XIST; miR-30b-5p; pneumonia.

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