Knockdown of MBD2 Attenuates LPS-Stimulated Inflammation and Apoptosis in WI-38 Cells Through the STAT-3 Pathway

Chen, Yao; Lu, Liqun

doi:10.2478/aite-2025-0021

Abstract

This study aims to investigate the role of MBD protein 2 (MBD2) in the pneumonia cell model of lipopolysaccharide (LPS)-stimulated WI-38 cells and to uncover the mechanism. LPS-stimulated WI-38 cells were constructed as an in vitro pneumonia model. Quantitative polymerase chain reaction (qPCR) and immunoblot assays showed MBD2 expression in WI-38 cells. Cell counting kit-8 (CCK-8) assays showed the growth of WI-38 cells. Flow cytometry assays showed the apoptosis of WI-38 cells after LPS treatment and siRNA transfection. Enzyme-linked immunosorbent assay (ELISA) and qPCR assays showed the effects on inflammation, and immunoblot assays further confirm the mechanism. MBD2 was highly expressed in LPS-stimulated WI-38 cells. Knockdown of MBD2 alleviates production of cellular inflammatory cytokines in LPS-stimulated WI-38 cells. Further, knockdown of MBD2 alleviates apoptosis in LPS-stimulated WI-38 cells. Mechanically, the knockdown of MBD2 regulates the signal transducer and activator of transcription (STAT)-3 pathway in LPS-stimulated WI-38 cells. Knockdown of MBD2 attenuates LPS-stimulated inflammation and apoptosis in WI-38 cells through the STAT-3 pathway. Therefore, MBD2 could serve as a promising target of pediatric pneumonia.

References

Cheng J, Song J, He X et al. (2016) Loss of Mbd2 protects mice against high-fat diet-induced obesity and insulin resistance by regulating the homeostasis of energy storage and expenditure. Diabetes 65:3384–3395. https://doi.org/10.2337/db16-0151
Search in Google Scholar Back to article
Chen Z, Shang Y, Yuan Y et al. (2022) MBD2 mediates Th17 cell differentiation by regulating MINK1 in Th17-dominant asthma. Front Genet 13:959059. https://doi.org/10.3389/fgene.2022.959059
Search in Google Scholar Back to article
de Benedictis FM, Kerem E, Chang AB et al. (2020) Complicated pneumonia in children. Lancet 396:786–798. https://doi.org/10.1016/S0140-6736(20)31550-6
Search in Google Scholar Back to article
Demaria M, Poli V (2012) Pro-malignant properties of STAT-3 during chronic inflammation. Oncotarget 3:359–360. https://doi.org/10.18632/oncotarget.482
Search in Google Scholar Back to article
Hojo K, Furuta T, Komaki S et al. (2023) Systemic inflammation caused by an intracranial mesenchymal tumor with a EWSR1:CREM fusion presenting associated with IL-6/STAT-3 signaling. Neuropathology 43:244–251. https://doi.org/10.1111/neup.12877
Search in Google Scholar Back to article
Jain S, Williams DJ, Arnold SR et al. (2015) Community-acquired pneumonia requiring hospitalization among U.S. children. N Engl J Med 372:835–845. https://doi.org/10.1056/NEJMoa1405870
Search in Google Scholar Back to article
Lei Q, Zhang W (2022) Methyl CpG binding domain protein 2 (MBD2) in inflammation. Chin Med J (Engl) 135:2880–2882. https://doi.org/10.1097/CM9.0000000000002482
Search in Google Scholar Back to article
Li R, Liang Q, Yang Q et al. (2024a) Hexahydrocurcumin from Zingiberis rhizoma attenuates lipopolysaccharide-induced acute pneumonia through JAK1/STAT-3 signaling pathway. Phytomedicine 122:155141. https://doi.org/10.1016/j.phymed.2023.155141
Search in Google Scholar Back to article
Li X, Wang J, Liao C et al. (2024b) The binding of PKCε and MEG2 to STAT-3 regulates IL-6-mediated microglial hyperalgesia during inflammatory pain. FASEB J 38:e23590. https://doi.org/10.1096/fj.202300152RR
Search in Google Scholar Back to article
Sathe PA, Dash M, Vaideeswar P et al. (2024) Pediatric pneumonia – a clinico-pathological study. Indian J Pathol Microbiol 67: 766–769. https://doi.org/10.4103/ijpm.ijpm_700_23
Search in Google Scholar Back to article
Sun T, Liu Q, Wang Y et al. (2021) MBD2 mediates renal cell apoptosis via activation of Tox4 during rhabdomyolysis-induced acute kidney injury. J Cell Mol Med 25:4562–4571. https://doi.org/10.1111/jcmm.16207
Search in Google Scholar Back to article
Tang D, Su W, Wang X et al. (2021) Clinicopathologic significance of MYD88 L265P mutation and expression of TLR4 and P-STAT-3 in primary central nervous system diffuse large B-cell lymphomas. Brain Tumor Pathol 38:50–58. https://doi.org/10.1007/s10014-020-00386-8
Search in Google Scholar Back to article
Turer RW, Gradwohl S, Stassun J et al. (2024) User-centered design and implementation of an interoperable FHIR application for pediatric pneumonia prognostication in a randomized trial. Appl Clin Inform 15:556–568. https://doi.org/10.1055/a-2297-9129
Search in Google Scholar Back to article
Wang B, Liu S, Fan B et al. (2018) PKM2 is involved in neuropathic pain by regulating ERK and STAT-3 activation in rat spinal cord. J Headache Pain 19:7. https://doi.org/10.1186/s10194-018-0836-4
Search in Google Scholar Back to article
Wang C, Zhang P, Li Y et al. (2023) Downregulation of TRIM27 alleviates hypoxic-ischemic encephalopathy through inhibiting inflammation and microglia cell activation by regulating STAT-3/HMGB1 axis. J Chem Neuroanat 129:102251. https://doi.org/10.1016/j.jchemneu.2023.102251
Search in Google Scholar Back to article
Wang Y, Zhang L, Wu GR et al. (2021) MBD2 serves as a viable target against pulmonary fibrosis by inhibiting macrophage M2 program. Sci Adv 7:eabb6075. https://doi.org/10.1126/sciadv.abb6075
Search in Google Scholar Back to article
Wu X, Wei S, Chen M et al. (2022b) P2RY13 exacerbates intestinal inflammation by damaging the intestinal mucosal barrier via activating IL-6/STAT-3 pathway. Int J Biol Sci 18:5056–5069. https://doi.org/10.7150/ijbs.74304
Search in Google Scholar Back to article
Wu GR, Zhou M, Wang Y et al. (2022a) Blockade of Mbd2 by siRNA-loaded liposomes protects mice against OVA-induced allergic airway inflammation via repressing M2 macrophage production. Front Immunol 13:930103. https://doi.org/10.3389/fimmu.2022.930103
Search in Google Scholar Back to article
Xiong C, Li B, Song R et al. (2024) IFITM3 mediates inflammation induced myocardial injury through JAK2/STAT-3 signaling pathway. Mol Immunol 167:1–15. https://doi.org/10.1016/j.molimm.2024.01.012
Search in Google Scholar Back to article
Zhao G, Jing J (2023) HOMER1A restores sevoflurane-induced cognitive dysfunction by regulating microglia’s activation through activating the AMPK/TXNIP axis. Signa Vitae 19:202–212. https://doi.org/10.22514/sv.2023.065
Search in Google Scholar Back to article

Knockdown of MBD2 Attenuates LPS-Stimulated Inflammation and Apoptosis in WI-38 Cells Through the STAT-3 Pathway

Abstract

Paradigm

My account