微小RNA-222-3p在儿童肺炎中的表达及其对LPS诱导的肺成纤维细胞凋亡和炎症因子表达的影响  

Expression of microRNA-222-3p in pediatric pneumonia and its effect on LPS-induced lung fibroblast apoptosis and inflammatory factor expression

作　　者：林报忠 郑通喜 周红艳[1] LIN Bao-zhong;ZHENG Tong-xi;ZHOU Hong-yan(Department of Neonatology,Sanya Maternal and Child Health Hospital,Sanya 572000,Hainan Province,China)

机构地区：[1]海南省三亚市妇幼保健院新生儿科,海南三亚572000

出　　处：《中国临床药理学杂志》2021年第12期1514-1517,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨微小RNA-222-3p(miR-222-3p)在肺炎患儿血清中的表达水平,以及对脂多糖(LPS)诱导的肺成纤维细胞中凋亡和炎症因子水平的影响。方法选取87例儿童血清样本(50例肺炎患儿为肺炎组,37例健康儿童为健康对照组)作为实验标本,用实时荧光定量聚合酶链反应(RT-q PCR)检测血清中miR-222-3p表达水平。将人胚肺成纤维细胞WI-38细胞株分为4组:对照组(无任何处理)、模型组(5μg·m L^(-1)LPS诱导12 h)、转染组-1(转染inhibitor NC后,5μg·m L^(-1)LPS诱导12 h后,)、转染组-1(转染miR-222-3p inhibitor后用5μg·m L^(-1)LPS诱导12 h)。用ELISA试剂盒检测各组WI-38细胞培养上清液白介素-1β(IL^(-1)β)、白介素6(IL-6)、肿瘤坏死因子-α(TNF-α)的水平;用流式细胞术比较各组细胞的凋亡率。结果健康对照组与肺炎组血清中miR-222-3p的相对表达分别为1.10±0.36,1.51±0.52;对照组、模型组、转染组-1、和转染组-2细胞上清IL^(-1)β的水平为(10.08±1.33),(317.85±37.67),(304.87±37.07),(152.12±24.38)pg·m L^(-1);IL-6的水平为(19.78±2.17),(423.75±54.16),(429.05±48.03),(208.18±34.85)pg·m L^(-1);TNF-α的含量为(17.54±3.85),(602.13±42.13)(606.44±53.32),(349.30±25.73)pg·m L^(-1);凋亡率分别为(4.73±0.64)%,(9.75±1.27)%,(10.68±1.87)%,(7.18±0.79)%;模型组与对照组比较,转染组-1与对照组比较,转染组-2与转染组-1比较,差异均有统计学意义(均P<0.05)。结论miR-222-3p在肺炎患儿血清中高表达,敲低miR-222-3p可降低LPS诱导的WI-38炎症因子水平,并抑制细胞凋亡。Objective To investigate the expression level of microRNA-222-3p(miR-222-3p)in serum of children with pneumonia and the role of miR-222-3p in lung fibroblasts induced by lipopolysaccharide(LPS).Methods Serum samples from 87 children(50 children with pneumonia in the pneumonia group and 37 healthy children in the healthy control group)were selected as experimental samples.Real-time quantitative polymerase chain reaction(RT-q PCR)was used to detect the expression level of miR-222-3p in serum.Human embryonic lung fibroblasts were assigned to four groups:Control group(without any treatment),Model group(induced by 5μg·m L^(-1)LPS for 12 h),Transfection-1 group(induced by 5μg·m L^(-1)LPS for 12 h after transfection with inhibitor NC),and Transfection-2 group(induced by 5μg·m L^(-1)LPS for 12 h after transfection with miR-222-3p inhibitor).The levels of interleukin-1β(IL^(-1)β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)in the supernatant of WI-38 cell culture were determined by ELISA kit.The apoptosis rate of each group was compared by flow cytometry.Results The relative expression of miR-222-3p in serum of healthy control group and pneumonia group was 1.10±0.36 and1.51±0.52,respectively.The levels of IL^(-1)βin the supernatant of control group,model group,transfected group-1 and transfected group-2 were(10.08±1.33),(317.85±37.67),(304.87±37.07),(152.12±24.38)pg·m L^(-1);IL-6 levels were(19.78±2.17),(423.75±54.16),(429.05±48.03),(208.18±34.85)pg·m L^(-1);The content of TNF-αwas(17.54±3.85),(602.13±42.13)(606.44±53.32),(349.30±25.73)pg·m L^(-1);Apoptosis rates were(4.73±0.64)%,(9.75±1.27)%,(10.68±1.87)%and(7.18±0.79)%,respectively.There were statistically significant differences between the model group and the control group,between the transfection group-1 and the control group,between the transfection group-2 and the transfection group-1(P<0.05).Conclusion miR-222-3p is highly expressed in the serum of children with pneumonia,and knockdown of miR-222-3p can reduce the level of LP

关 键 词：儿童肺炎 miR-222-3p WI-38细胞 凋亡 炎症因子 

分 类 号：R97[医药卫生—药品]