LncRNA XIST调节miR-574-5p/TLR4轴对呼吸道合胞病毒感染的支气管上皮细胞凋亡的影响

付晓康

文章摘要

LncRNA XIST调节miR-574-5p/TLR4轴对呼吸道合胞病毒感染的支气管上皮细胞凋亡的影响

Effect of LncRNA XIST on apoptosis of bronchial epithelial cells infected with respiratory syncytial virus by regulating the miR-574-5p/TLR4 axis

投稿时间：2023-12-29 修订日期：2024-04-10

DOI：

中文关键词: LncRNA XIST miR-574-5p/TLR4轴呼吸道合胞病毒支气管上皮细胞凋亡

英文关键词: LncRNA XIST MiR-574-5p/TLR4 axis Respiratory syncytial virus Bronchial epithelial cells Apoptosis

基金项目:2023年市级科技计划自筹经费项目（NO.2322087D）

作者	单位	邮编
付晓康^*	河北北方学院附属第二医院	075000

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中文摘要:

目的:探讨长链非编码RNA（LncRNA）XIST调节miR-574-5p/TLR4轴对呼吸道合胞病毒（RSV）感染的支气管上皮细胞凋亡的影响。方法:将人支气管上皮细胞（16HBE）分为control组、RSV组、si-NC组（RSV感染）、si-XIST组（RSV感染）、si-XIST+inhibitor-NC组（RSV感染）、si-XIST+miR-574-5p inhibitor组（RSV感染），实时荧光定量PCR检测LncRNA XIST、miR-574-5p表达；EdU染色、流式细胞术分别检测细胞增殖、凋亡；ELISA检测肿瘤坏死因子（TNF-α）、白细胞介素-6、-1β（IL-6、IL-1β）的分泌；双荧光素酶报告基因实验验证miR-574-5p与LncRNA XIST和TLR4的关系；Western blot检测TLR4、增殖细胞核抗原（PCNA）、Bcl-2相关X蛋白（Bax）、细胞抗凋亡因子B淋巴细胞瘤-2（Bcl-2）、切割型半胱氨酸天冬氨酸蛋白水解酶-3（Cleaved Caspase-3）蛋白表达。结果：与control组比较，RSV组、si-NC组XIST、凋亡率、IL-6、IL-1β、TNF-α水平、TLR4、Bax、Cleaved Caspase-3表达升高，Edu阳性率、miR-574-5p、PCNA、Bcl-2表达降低（P<0.05）；与si-NC组比较，si-XIST组XIST表达、凋亡率、IL-6、IL-1β、TNF-α水平、TLR4、Bax、Cleaved Caspase-3表达降低，Edu阳性率、miR-574-5p、PCNA、Bcl-2表达升高（P<0.05）；下调miR-574-5p可逆转敲低XIST对RSV感染的16HBE细胞凋亡和炎症反应的抑制（P<0.05）；双荧光素酶报告基因实验证实miR-574-5p与LncRNA XIST、miR-574-5p与TLR4存在靶向调控关系（P<0.05）。结论：LncRNA XIST在RSV感染的16HBECs细胞中上调表达，敲低LncRNA XIST可能通过调节miR-574-5p/TLR4轴，抑制RSV感染后的16HBECs细胞凋亡。

英文摘要:

Objective: To investigate the effect of long non-coding RNA (LncRNA) XIST on apoptosis of bronchial epithelial cells infected with respiratory syncytial virus (RSV) by regulating the miR-574-5p/TLR4 axis. Methods: Human bronchial epithelial cells (16HBE) were separated into control group, RSV group, si-NC group (RSV infection), si-XIST group (RSV infection), si-XIST+inhibitor NC group (RSV infection), and si-XIST+miR-574-5p inhibitor group (RSV infection), Real-time fluorescence quantitative PCR was applied to detect the expression of LncRNA XIST and miR-574-5p; EdU staining and flow cytometry were applied to detect cell proliferation and apoptosis, respectively; ELISA was applied to detect the secretions of tumor necrosis factor-α (TNF-α), interleukin-6, and interleukin-1β (IL-6, IL-1β); dual luciferase reporter gene experiment was applied to verify the relationship between miR-574-5p and LncRNA XIST, TLR4; Western blot was applied to detect the expression of TLR4, proliferating cell nuclear antigen (PCNA), Bcl-2 associated X protein (Bax), anti apoptotic factor B cell lymphomatoma-2 (Bcl-2), and Cleaved caspase-3 proteins. Results: Compared with the control group, the XIST, apoptosis rate, IL-6, IL-1β, TNF-α levels, TLR4, Bax, and Cleaved Caspase-3 expression in RSV group and si-NC group elevated, the positive rate of Edu, miR-574-5p, PCNA, and Bcl-2 expression decreased (P<0.05); compared with the si-NC group, the XIST, apoptosis rate, IL-6, IL-1β, TNF-α levels, TLR4, Bax, and Cleaved Caspase-3 expression in the si-XIST group decreased, the positive rate of Edu, miR-574-5p, PCNA, and Bcl-2 expression increased (P<0.05); downregulation of miR-574-5p was able to reverse the inhibitory effects of knocking down XIST on apoptosis and inflammatory response of 16HBE cells infected with RSV (P<0.05); dual luciferase reporter gene experiment confirmed the targeted regulatory relationship between miR-574-5p and LncRNA XIST, and between miR-574-5p and TLR4 (P<0.05). Conclusion: LncRNA XIST is upregulated in RSV infected 16HBECs cells, and knocking down LncRNA XIST may inhibit apoptosis of 16HBECs cells after RSV infection by regulating the miR-574-5p/TLR4 axis.

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