Downregulation of circ_0035292 Alleviates LPS-Induced WI-38 Cell Injury via Targeting miR-494-3p/TLR4 Pathway in Infantile Pneumonia.

Circular RNAs (circRNAs) have been confirmed to mediate infantile pneumonia development. In this, we investigated the role and new mechanism of circ_0035292 regulating infantile pneumonia progression. Lipopolysaccharide (LPS)-treated WI-38 cells were used to mimic infantile pneumonia cell injury models. Quantitative real-time PCR was used to measure circ_0035292, microRNA (miR)-494-3p and toll-like receptor 4 (TLR4). Cell proliferation and apoptosis were assessed by MTT assay, EdU assay, and flow cytometry. Protein expression was tested using western blot analysis. Inflammation and oxidative stress were evaluated by measuring IL-6, IL-1β, MDA and SOD levels using ELISA assay and corresponding kits. RNA interaction was confirmed by dual-luciferase reporter assay and RIP assay. Circ_0035292 had elevated expression in infantile pneumonia patients and LPS-induced WI-38 cells. Silenced circ_0035292 could enhance WI-38 cell proliferation, while suppress apoptosis, inflammation and oxidative stress under LPS treatment. Mechanically, circ_0035292 targeted miR-494-3p to positively regulate TLR4. The rescue experiments indicated that miR-494-3p inhibitor abolished the function of circ_0035292 knockdown, and TLR4 overexpression reversed the inhibitory effect of miR-494-3p on LPS-induced WI-38 cell injury. Circ_0035292 might be a potential target for infantile pneumonia treatment, which knockdown could relieve LPS-induced cell injury via the regulation of miR-494-3p/TLR4 axis.

Dai Z ，Hu J ，Luo Z ，Xiao J ... -  《-》

Circ_0026579 alleviates LPS-induced WI-38 cells inflammation injury in infantile pneumonia.

Yu Y ，Yang T ，Ding Z ，Cao Y ... -  《-》

Circ-BICC1 Knockdown Alleviates Lipopolysaccharide (LPS)-Induced WI-38 Cell Injury Through miR-338-3p/MYD88 Axis.

Circular RNAs (circRNAs) play important roles in human diseases, including infantile pneumonia. In this article, we aimed to investigate the functions of circ-BICC1 in lipopolysaccharide (LPS)-induced injury of WI-38 cells. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed for the levels of circ-BICC1, BICC1, microRNA-338-3p (miR-338-3p), and myeloid differentiation primary response 88 (MYD88). Cell Counting Kit-8 (CCK-8) assay, 5-ethynyl-2'-deoxyuridine (EdU) assay, and flow cytometry analysis were conducted to evaluate cell viability, proliferation, and apoptosis, respectively. Enzyme-linked immunosorbent assay (ELISA) kits were used for the concentrations of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). The levels of oxidative stress markers were detected with commercial kits. Dual-luciferase reporter assay was adopted to analyze the interaction between circ-BICC1 and miR-338-3p, as well as MYD88 and miR-338-3p. Western blot assay was employed for the protein level of MYD88. Circ-BICC1 level was increased in pneumonia patients' blood samples and LPS-treated WI-38 cells. LPS treatment suppressed WI-38 cell viability and promoted cell apoptosis, inflammation, and oxidative stress. Circ-BICC1 knockdown reversed the effect of LPS-induced WI-38 cell injury. For mechanism analysis, circ-BICC1 could function as the sponge for miR-338-3p and miR-338-3p inhibition reversed the effect of circ-BICC1 knockdown on LPS-induced WI-38 cell injury. MYD88 was identified as the target of miR-338-3p. MiR-338-3p overexpression relieved LPS-induced injury of WI-38 cells, while the impact was abolished by elevating MYD88. Circ-BICC1 silencing remitted LPS-triggered WI-38 cell damage by adsorbing miR-338-3p and regulating MYD88.

Wang J ，Li G ，Lin S ，Cheng L ... -  《-》

Circ_ZNF652 regulates the proliferation and apoptosis of LPS-induced WI-38 cells via miR-302e/TLR4 axis.

Circular RNAs (circRNAs) play an important regulatory role in multiple human diseases, including organ allograft rejection. Infantile pneumonia (IP) is a common disease that seriously threatens the health of infants and young children. CircRNAs have been shown to be involved in the advance of IP. However, the function of circ_ZNF652 in IP has not been fully studied. Lipopolysaccharide (LPS)-treated WI-38 cells were used as cell injury models of IP. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of circ_ZNF652, miR-302e and Toll-like receptor 4 (TLR4). 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium Bromide (MTT) assay, thymidine analog 5-ethynyl-2'-deoxyuridine (EdU) assay, and flow cytometry assay were utilized to explore cell functions. Western blot was employed to examine the protein levels of PCNA, Bcl-2, Bax, and TLR4. ELISA was used to detect the release of inflammatory cytokines. The relationship between miR-302e and circ_ZNF652 or TLR4 was verified by dual-luciferase reporter assay and RNA pull down assay. Circ_ZNF652 was significantly up-regulated in serum of IP patients and LPS-induced WI-38 cells. Silencing circ_ZNF652 enhanced cell proliferation and inhibited cell apoptosis in LPS-induced WI-38 cells. MiR-302e was identified as a target of circ_ZNF652, and knockdown of circ_ZNF652 alleviated LPS-induced WI-38 cell injuries by up-regulating miR-302e. In addition, TLR4 was a downstream target of miR-302e. Overexpression of TLR4 recovered cell apoptosis and inflammation that were repressed by miR-302e enrichment in LPS-induced WI-38 cells. Circ_ZNF652 regulates the expression of TLR4 by regulating miR-302e, thereby mediating cell proliferation, apoptosis and inflammation. The results provide a novel targeted therapy for IP.

Wang H ，Wang Q 《-》

Circ_0099630 knockdown alleviates lipopolysaccharide-induced injuries of human periodontal ligament cells through the inhibition of TLR4 by releasing miR-409-3p.

Periodontitis triggers tooth loss and affects the health of population worldwide. Emerging evidence hints that circular RNAs (circRNAs) are involved in various diseases, including periodontitis. This study aimed to investigate the role of circ_0099630 in the progression of periodontitis. Periodontitis cell model was constructed by treating human periodontal ligament cells (HPDLCs) with lipopolysaccharide (LPS). Quantitative real-time PCR was used to analyze the expression of circ_0099630, microRNA-409-3p (miR-409-3p) and toll-like receptor 4 (TLR4) mRNA. Western blot was used for detecting protein levels of TLR4, cleaved-caspase 3, Bcl-2, CyclinD1 and NF-κB signaling markers. For function analyses, cell proliferation was assessed by CCK-8 assay and EdU assay. The releases of pro-inflammation factors were monitored by ELISA kits. The potential relationship between miR-409-3p and circ_0099630 or TLR4 was verified by dual-luciferase reporter assay, RIP assay and pull-down assay. The expression of circ_0099630 and TLR4 was elevated in periodontitis patients and LPS-treated HPDLCs. LPS induced HPDLC proliferation inhibition, apoptosis and inflammatory responses, while circ_0099630 knockdown or TLR4 knockdown alleviated these injuries. Besides, TLR4 overexpression reversed the inhibitory effect of circ_0099630 knockdown on LPS-induced HPDLC injuries. Mechanism analysis showed that circ_0099630 positively regulated TLR4 expression by acting as miR-409-3p sponge. MiR-409-3p restoration largely ameliorated LPS-induced HPDLC injuries by depleting TLR4. Moreover, LPS activated the NF-κB signaling pathway, while circ_0099630 knockdown inhibited the activity of NF-κB signaling via the miR-409-3p/TLR4 axis. Circ_0099630 knockdown relieved LPS-induced HPDLC injury by miR-409-3p/TLR4 axis, suggesting that circ_0099630 might be a potential target for periodontitis treatment.

Qi H ，Han B ，Che J 《BMC Oral Health》