Department of Pediatrics, Henan Provincial People's Hospital, Zhengzhou, China.
Artif Cells Nanomed Biotechnol. 2020 Dec;48(1):498-505. doi: 10.1080/21691401.2020.1716781.
Infantile pneumonia (IP) seriously affects the health of children. This article mainly discussed the protective effect of long non-coding RNA NKILA (lnc NKILA) on IP by detecting cell viability, apoptosis and inflammatory response of MRC5 cells. Cell counting kit-8 (CCK-8) was used to detect cell viability, while flow cytometry was used to detect cell apoptosis. The expression of apoptosis-associated factors (Bcl-2, Bax, PARP and Cleaved-PARP) and NF-κB and JNK pathway-related factors (t-IκBα, p-IκBα, t-p65, p-p65, β-actin, t-JNK and p-JNK) were tested by western blot. Otherwise, productions of inflammatory factors interleukin (IL)-1β and IL-6 were tested by enzyme-linked immunosorbent assay (ELISA) and western blot. Furthermore, RNA levels were respectively tested and changed by RT-qPCR and cell transfection. Tumour necrosis factor-α (TNF-α) treatment reduced cell viability, induced cell apoptosis and promoted inflammatory factors expression. NKILA overexpression remitted TNF-α-induced injury. Moreover, NKILA positively regulated miR-21. miR-21 inhibition could weaken the functions of NKILA overexpression on TNF-α-induced injury. At last, NKILA and miR-21 were involved in the regulation of JNK and NF-κB pathways. NKILA overexpression remitted TNF-α-induced MRC5 cell injury by up-regulation of miR-21 and via inactivation of JNK and NF-κB signaling pathways.
婴儿肺炎（IP）严重影响儿童健康。本文主要通过检测 MRC5 细胞的活力、凋亡和炎症反应，探讨长链非编码 RNA NKILA（lnc NKILA）对 IP 的保护作用。细胞计数试剂盒-8（CCK-8）用于检测细胞活力，流式细胞术用于检测细胞凋亡。通过 Western blot 检测凋亡相关因子（Bcl-2、Bax、PARP 和 Cleaved-PARP）和 NF-κB 和 JNK 通路相关因子（t-IκBα、p-IκBα、t-p65、p-p65、β-actin、t-JNK 和 p-JNK）的表达。此外，通过酶联免疫吸附试验（ELISA）和 Western blot 检测炎症因子白细胞介素（IL）-1β和 IL-6 的产生。进一步通过 RT-qPCR 和细胞转染分别检测 RNA 水平并改变。肿瘤坏死因子-α（TNF-α）处理降低细胞活力，诱导细胞凋亡，促进炎症因子表达。NKILA 过表达缓解 TNF-α诱导的损伤。此外，NKILA 正向调控 miR-21。miR-21 抑制可减弱 NKILA 过表达对 TNF-α诱导损伤的作用。最后，NKILA 和 miR-21 参与 JNK 和 NF-κB 通路的调节。NKILA 过表达通过上调 miR-21 缓解 TNF-α诱导的 MRC5 细胞损伤，并通过失活 JNK 和 NF-κB 信号通路。
