Department of Pediatrics, Zhengzhou Central Hospital, Zhengzhou University, Zhengzhou, Henan, China.
Autoimmunity. 2022 Jun;55(4):233-242. doi: 10.1080/08916934.2022.2062595. Epub 2022 Apr 28.
Circular RNAs (circRNAs) have been reported to be involved in the progression of infantile pneumonia. Here, we investigated the function of circTRHDE in lipopolysaccharide (LPS)-induced cell inflammatory injury to evaluate its role in infantile pneumonia progression.
The circTRHDE, microRNA (miR)-381-3p and TNF-receptor associated factor 3 (TRAF3) expression were detected by quantitative real-time PCR. LPS-induced WI-38 cells were used to construct an inflammatory injury model. Cell viability, inflammation and apoptosis were measured by cell counting kit assay, ELISA assay and flow cytometry. Caspase3 activity, MDA level and SOD activity were analysed to assess cell apoptosis and oxidative stress. Protein levels were determined using western blot analysis. The interaction between miR-381-3p and circTRHDE or TRAF3 was confirmed by dual-luciferase activity assay and RNA pull-down assay.
CircTRHDE had increased expression in infantile pneumonia patients and LPS-induced WI-38 cells. LPS treatment inhibited WI-38 cell viability while promoting inflammation, apoptosis and oxidative stress. However, knockdown of circTRHDE remitted LPS-triggered WI-38 cell injury. CircTRHDE could sponge miR-381-3p to positively regulate TRAF3 expression. MiR-381-3p suppressed LPS-induced WI-38 cell inflammatory injury, and this effect was revoked by TRAF3 overexpression. Also, LPS-induced WI-38 cell inflammatory injury restrained by circTRHDE knockdown also were reversed by miR-318-3p inhibitor or TRAF3 overexpression.
Our findings demonstrated that circTRHDE might be a target for infantile pneumonia treatment, which relieved LPS-induced cell inflammatory injury by the regulation of the miR-318-3p/TRAF3 axis.
环状 RNA（circRNAs）已被报道参与婴儿肺炎的进展。在这里，我们研究了 circTRHDE 在脂多糖（LPS）诱导的细胞炎症损伤中的功能，以评估其在婴儿肺炎进展中的作用。
通过实时定量 PCR 检测 circTRHDE、microRNA（miR）-381-3p 和 TNF 受体相关因子 3（TRAF3）的表达。用 LPS 诱导 WI-38 细胞构建炎症损伤模型。用细胞计数试剂盒检测细胞活力，ELISA 检测炎症因子，流式细胞术检测细胞凋亡。用 caspase3 活性、MDA 水平和 SOD 活性分析评估细胞凋亡和氧化应激。用 Western blot 分析测定蛋白水平。通过双荧光素酶活性测定和 RNA 下拉实验证实 miR-381-3p 与 circTRHDE 或 TRAF3 的相互作用。
circTRHDE 在婴儿肺炎患者和 LPS 诱导的 WI-38 细胞中表达增加。LPS 处理抑制 WI-38 细胞活力，同时促进炎症、凋亡和氧化应激。然而，circTRHDE 的敲低缓解了 LPS 触发的 WI-38 细胞损伤。circTRHDE 可以吸附 miR-381-3p 来正向调节 TRAF3 的表达。miR-381-3p 抑制 LPS 诱导的 WI-38 细胞炎症损伤，而 TRAF3 过表达则取消了这一作用。同样，circTRHDE 敲低缓解的 LPS 诱导的 WI-38 细胞炎症损伤也被 miR-318-3p 抑制剂或 TRAF3 过表达所逆转。
我们的研究结果表明，circTRHDE 可能是治疗婴儿肺炎的一个靶点，它通过调节 miR-318-3p/TRAF3 轴缓解 LPS 诱导的细胞炎症损伤。
