Abstract
Objective: Mounting studies have clarified the link between long non-coding RNAs (lncRNAs) and pneumonia. This research aims to probe the function and regulatory mechanism of lncRNA H19 in lipopolysaccharide (LPS)-induced cell models of pneumonia.
Methods: WI-38 cells were exposed to LPS for 12 h to mimic cell models of pneumonia. The relative expression of H19, miR-140-5p, and toll-like receptor 4 (TLR4) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The cell viability was detected by MTT assay. The protein expression of apoptosis-associated proteins (Bax and Bcl-2) and TLR4 were determined by western blot. Moreover, the content of interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α were measured by enzyme-linked immunosorbent assay (ELISA). The target relationship between miR- 140-5p and H19/ TLR4 was confirmed by Dual luciferase reporter (DLR) assay.
Results: LncRNA H19 and TLR4 were up-regulated, while miR-140-5p was downregulated in peripheral blood of patients with pneumonia and LPS-treated WI-38 cells compared with their controls. Silencing of H19 or miR-140-5p mimics facilitated cell viability, whereas repressed apoptosis and reduced content of TNF-α, IL-6, and IL-1β in LPS-induced WI-38 cells. H19 targeted miR-140-5p and it inversely regulated miR-140- 5p expression. MiR-140-5p targeted TLR4 and it inversely regulated TLR4 expression. H19 positively regulated TLR4 expression. Moreover, inhibition of miR-140-5p or overexpression of TLR4 reversed the effects of H19 silencing on cell viability, inflammation, and apoptosis in LPS-induced WI-38 cells.
Conclusion: Silencing of H19 inhibited apoptosis and inflammation by miR-140- 5p/TLR4 pathway in LPS-induced WI-38 cells.
Keywords: Pneumonia, H19, miR-140-5p, TLR4, inflammation, apoptosis.
[PMID: 17968291]
[http://dx.doi.org/10.1016/j.rcl.2011.06.007] [PMID: 21889014]
[PMID: 16477891]
[http://dx.doi.org/10.1016/j.biocel.2006.06.001] [PMID: 16844403]
[http://dx.doi.org/10.1016/S0140-6736(13)60648-0] [PMID: 23582723]
[http://dx.doi.org/10.1111/irv.12738] [PMID: 32267084]
[http://dx.doi.org/10.1002/14651858.CD009576.pub2] [PMID: 24859388]
[http://dx.doi.org/10.36290/vnl.2017.105] [PMID: 28933177]
[http://dx.doi.org/10.1007/978-981-10-5203-3_1] [PMID: 28815535]
[http://dx.doi.org/10.1186/1480-9222-16-11] [PMID: 25276098]
[http://dx.doi.org/10.1016/j.lfs.2019.05.008] [PMID: 31071307]
[http://dx.doi.org/10.1002/cbf.3392] [PMID: 31066476]
[http://dx.doi.org/10.1007/s12031-020-01751-0] [PMID: 33205379]
[http://dx.doi.org/10.1080/01902148.2021.1887967] [PMID: 33629893]
[http://dx.doi.org/10.1007/978-1-4939-3378-5_21] [PMID: 26721498]
[http://dx.doi.org/10.1111/jcmm.13866] [PMID: 30188595]
[http://dx.doi.org/10.1038/nrm1644] [PMID: 15852042]
[http://dx.doi.org/10.1126/science.1076311] [PMID: 12242443]
[http://dx.doi.org/10.1073/pnas.1818522116] [PMID: 30971494]
[http://dx.doi.org/10.1080/23744235.2019.1671987] [PMID: 31583932]
[http://dx.doi.org/10.1002/jcp.27642] [PMID: 30367500]
[http://dx.doi.org/10.3892/etm.2018.6692] [PMID: 30344669]
[http://dx.doi.org/10.1016/j.ccm.2004.10.014] [PMID: 15802172]
[http://dx.doi.org/10.1016/j.lfs.2018.11.056] [PMID: 30500550]
[http://dx.doi.org/10.1016/j.biopha.2018.04.192] [PMID: 29864958]
[http://dx.doi.org/10.1378/chest.11-1160] [PMID: 22406959]
[http://dx.doi.org/10.1016/S0140-6736(06)69334-3] [PMID: 16997649]
[http://dx.doi.org/10.1177/0963689720918294] [PMID: 32308025]
[http://dx.doi.org/10.1186/s12931-018-0956-z] [PMID: 30547791]
[http://dx.doi.org/10.1111/j.1582-4934.2007.00131.x] [PMID: 18419599]
[http://dx.doi.org/10.1046/j.1440-1711.2001.00980.x] [PMID: 11264706]
[http://dx.doi.org/10.2147/OTT.S245710] [PMID: 32425552]
[http://dx.doi.org/10.21037/apm-20-929] [PMID: 32576007]
[http://dx.doi.org/10.1038/srep31663] [PMID: 27526865]
[http://dx.doi.org/10.1007/s10753-018-0741-7] [PMID: 29508184]
[http://dx.doi.org/10.1016/j.ajpath.2011.09.019] [PMID: 22062220]
[http://dx.doi.org/10.1002/2211-5463.12322] [PMID: 29226075]