长链非编码RNA NNT-AS1在肺炎患儿血清中的表达及其对脂多糖诱导的肺上皮细胞损伤的影响  

Expression of lncRNA NNT-AS1 in serum of children with pneumonia and its effect on LPS-induced lung epithelial cell injury

作　　者：王淑玉[1] 付德龙[1] 李存宇[1] WANG Shuyu;FU Delong;LI Cunyu(Department of Neonatology,Tengzhou Central People's Hospital,Tengzhou Shandong 277599,China)

机构地区：[1]滕州市中心人民医院新生儿科,山东滕州277599

出　　处：《中国感染与化疗杂志》2023年第2期143-148,共6页Chinese Journal of Infection and Chemotherapy

摘　　要：目的 探究长链非编码RNA(lncRNA) NNT-AS1在肺炎患儿血清中的表达及其对脂多糖(LPS)诱导的肺上皮细胞损伤的影响。方法 实时荧光定量PCR(qRT-PCR)检测NNT-AS1相对表达水平。将肺泡上皮细胞BEAS-2B分为对照组、LPS组(1 mg/L的LPS处理细胞)、LPS+si-NC组(转染si-NC,LPS处理细胞)、LPS+si-NNT-AS1组(转染si-NNTAS1,LPS处理细胞)、LPS+si-NNT-AS1+SB203580组(转染si-NNT-AS1,LPS处理和MAPK信号通路抑制剂SB203580细胞)。qRT-PCR检测NNT-AS1相对表达水平,流式细胞术检测细胞凋亡,蛋白质印迹法(Western blot)检测cleaved-caspase3、Bax、p-ERK1/2、p-p38 MAPK蛋白表达,ELISA检测IL-6、IL-1β、TNF-α。结果 NNT-AS1在肺炎患儿血清中的相对表达水平高于健康对照。与对照组相比,LPS组中NNT-AS1相对表达水平、凋亡率、cleaved-caspase3、Bax、p-ERK1/2、p-p38MAPK蛋白表达水平明显增加,IL-6、IL-1β、TNF-α增多。与LPS+si-NC组相比,LPS+si-NNT-AS1组NNT-AS1相对表达水平、凋亡率、cleaved-caspase3、Bax、p-ERK1/2、p-p38 MAPK蛋白表达水平明显降低,IL-6、IL-1β、TNF-α减少。与LPS+si-NNT-AS1组相比,LPS+si-NNT-AS1+SB203580组凋亡率、cleaved-caspase3、Bax蛋白表达水平明显降低,IL-6、IL-1β、TNF-α含量减少。结论 沉默NNT-AS1可能通过抑制MAPK信号通路减轻LPS诱导的肺泡上皮凋亡和炎症反应。Objective To investigate the expression of long non-coding RNA(lncRNA) NNT-AS1 in serum of children with pneumonia and its effect on lipopolysaccharide(LPS)-induced lung epithelial cell injury. Methods Alveolar epithelial cells BEAS-2B were divided into control group, LPS group(1 mg/L LPS-treated cells), LPS+si-NC group(si-NC transfected, LPS-treated cells),LPS+si-NNT-AS1 group(transfected si-NNT-AS1, LPS-treated cells), LPS+si-NNT-AS1+ inhibitor group(transfected si-NNTAS1, LPS-treated and MAPK signaling pathway inhibitor SB203580 cells). The relative expression level of NNT-AS1 was detected by real-time quantitative PCR(qRT-PCR). The apoptosis was detected by flow cytometry. The protein expressions of cleaved caspase-3, Bax, p-ERK1/2 and p-p38 MAPK were detected by Western blot. IL-6, IL-1β and TNF-α were analyzed by enzyme-linked immunosorbent assay(ELISA). Results The relative expression level of NNT-AS1 in children with pneumonia was higher than that in healthy controls. Compared with the control group, LPS group showed significantly higher relative expression level of NNT-AS1,higher apoptosis rate, higher expression levels of cleaved caspase-3, Bax, p-ERK1/2, and p-p38 MAPK protein, and higher levels of IL-6, IL-1β, TNF-α. Compared with LPS+si-NC group, LPS+si-NNT-AS1 group was associated with significantly decreased relative expression level of NNT-AS1, lower apoptosis rate, lower expression levels of cleaved caspase-3, Bax, p-ERK1/2 and p-p38 MAPK,and lower levels of IL-6, IL-1β and TNF-α. Compared with LPS+si-NNT-AS1 group, LPS+si-NNT-AS1+SB203580 group had significantly decreased apoptosis rate, lower expression levels of cleaved caspase-3, and Bax protein, and decreased levels of IL-6, IL-1β, TNF-α. Conclusions Silencing NNT-AS1 may alleviate LPS-induced alveolar epithelial apoptosis and inflammatory response by inhibiting MAPK signaling pathway.

关 键 词：长链非编码RNA NNT-AS1 脂多糖 肺上皮细胞 凋亡 炎症反应 

分 类 号：R563.1[医药卫生—呼吸系统]