Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University, Nanjing, Jiangsu, China.
Department of Periodontics, The Affiliated Stomatological Hospital of Nanjing Medical University, Nanjing, Jiangsu, China.
Histol Histopathol. 2020 Dec;35(12):1473-1481. doi: 10.14670/HH-18-278. Epub 2020 Nov 4.
Ulcerative colitis (UC) is a risk factor for carcinogenesis of colorectal cancer, which is associated with disruption of the epithelial barrier and disorder of the inflammatory response. It has been reported that the expression of microRNA (miR)-215 is upregulated in patients with long-term UC. The present study aimed to investigate the effects of miR-215 on lipopolysaccharide (LPS)-induced inflammatory injury in CCD-18Co cells, as well as to identify the underlying possible molecular mechanisms. CCD-18Co cells were treated with 1 µg/ml LPS to induce inflammatory injury. Reverse transcription-quantitative PCR was performed to determine the expression of miR-215 in LPS-treated CCD-18Co cells. Moreover, a dual luciferase reporter system assay was used to evaluate the interaction of miR-215 and growth differentiation factor 11 (GDF11) in CCD-18Co cells. The expression of miR-215 was significantly upregulated in LPS-treated CCD-18Co cells. Knockdown of miR-215 significantly alleviated the inflammatory response and oxidative stress in LPS-treated CCD-18Co cells. In addition, GDF11 was identified as a direct binding target of miR-215 in CCD-18Co cells. Knockdown of miR-215 significantly increased the expression of GDF11, but decreased the expression levels of Toll-like receptor (TLR)4, phosphorylated (p)-p65, iNOS, p-p38 and p-JNK in LPS-treated CCD-18Co cells. Collectively, the present findings indicated that knockdown of miR-215 alleviated oxidative stress and inflammatory response in LPS-treated CCD-18Co cells by upregulating GDF11 expression and inactivating the TLR4/NF-κB and JNK/p38 signaling pathways.
溃疡性结肠炎(UC)是结直肠癌发生的致癌因素,与上皮屏障破坏和炎症反应紊乱有关。据报道,miR-215 在长期 UC 患者中的表达上调。本研究旨在探讨 miR-215 对 CCD-18Co 细胞脂多糖(LPS)诱导的炎症损伤的影响,并确定潜在的可能分子机制。用 1μg/ml LPS 处理 CCD-18Co 细胞以诱导炎症损伤。采用逆转录定量 PCR 检测 LPS 处理的 CCD-18Co 细胞中 miR-215 的表达。此外,采用双荧光素酶报告基因系统检测 miR-215 与 CCD-18Co 细胞中生长分化因子 11(GDF11)的相互作用。LPS 处理的 CCD-18Co 细胞中 miR-215 的表达明显上调。敲低 miR-215 可显著减轻 LPS 处理的 CCD-18Co 细胞的炎症反应和氧化应激。此外,GDF11 被鉴定为 CCD-18Co 细胞中 miR-215 的直接结合靶标。敲低 miR-215 可显著增加 GDF11 的表达,同时降低 LPS 处理的 CCD-18Co 细胞中 TLR4、磷酸化(p)-p65、iNOS、p-p38 和 p-JNK 的表达水平。综上所述,研究结果表明,通过上调 GDF11 的表达和抑制 TLR4/NF-κB 和 JNK/p38 信号通路,敲低 miR-215 可减轻 LPS 处理的 CCD-18Co 细胞中的氧化应激和炎症反应。