miRNA-432-5p在脂多糖诱导的肺泡上皮细胞损伤中的作用及可能机制研究  

Role of miRNA-432-5p in alveolar epithelial cells injury induced by lipopolysaccharide and its possible mechanism

作　　者：吴畏[1] 杨飞云[1] 牛丽丹[1] 石金河[1] WU Wei;YANG Fei-yun;NIU Li-dan;SHI Jin-he(Department of Emergency Medicine,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan,CHINA)

机构地区：[1]新乡医学院第一附属医院急救医学科,河南卫辉453100

出　　处：《海南医学》2024年第11期1521-1526,共6页Hainan Medical Journal

基　　金：国家自然科学基金青年基金(编号:81502313)。

摘　　要：目的探讨miRNA-432-5p在脂多糖(LPS)诱导的肺泡上皮细胞损伤中的作用及可能机制。方法人肺泡上皮细胞A549随机分为对照组、LPS组、miRNA-432-5p mimics组及miRNA-432-5p siRNA组。利用CCK-8实验检测细胞的活性;利用ELISA法检测细胞培养液中炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β)的含量;利用RT-qPCR检测细胞中miRNA-432-5p mRNA的表达;利用Hoechst染色检测细胞的凋亡情况;利用Western blot检测细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、半胱氨酸蛋白酶1(Caspase1)、白细胞介素1β(IL-1β)蛋白的表达。结果LPS组细胞的活性OD值(6 h:0.809±0.09;12 h:0.601±0.09;24 h:0.485±0.43)明显低于对照组的1.000±0.00,差异均具有统计学意义(P<0.05);LPS组细胞中miRNA-432-5p mRNA表达水平为1.563±0.10,明显高于对照组的1.000±0.00,细胞凋亡率为(41.52±1.83)%,明显高于对照组的(0.000±0.00)%,差异均有统计学意义(P<0.05);LPS组培养液中的TNF-α、IL-6和IL-1β含量分别为(36.32±6.07)pg/mL、(45.08±3.75)pg/mL、(40.87±6.65)pg/mL,明显高于对照组的(5.27±0.84)pg/mL、(5.57±0.65)pg/mL、(3.26±0.85)pg/mL,差异均具有统计学意义(P<0.05),LPS组细胞中的NLRP3、Caspase1和IL-1β蛋白表达水平分别为0.734±0.08、0.305±0.06、0.430±0.05,明显高于对照组的0.163±0.03、0.025±0.01、0.032±0.01,差异均具有统计学意义(P<0.05)。miRNA-432-5p mimics组细胞凋亡率为(24.23±3.09)%,明显低于LPS组的(41.52±1.83)%,细胞活性为0.639±0.09,明显高于LPS组的0.468±0.07,差异均有统计学意义(P<0.05);miRNA-432-5p mimics组培养液中的TNF-α、IL-6和IL-1β含量分别为(22.85±4.01)pg/mL、(29.15±5.22)pg/mL、(20.63±3.89)pg/mL,明显低于LPS组的(36.32±6.07)pg/mL、(45.08±3.75)pg/mL、(40.87±6.65)pg/mL,差异均具有统计学意义(P<0.05);miRNA-432-5p mimics组细胞中的NLRP3、Caspase1和IL-1β蛋白表达水平分别为0.473±0.04、0.121±0.02、0.279±0.Objective To investigate the role and possible mechanism of miRNA-432-5p in alveolar epithelial cells injury induced by LPS.Methods Human alveolar epithelial A549 cells were divided into 4 groups randomly:control group,LPS group,miRNA-432-5p mimics group,and miRNA-432-5p siRNA group.CCK-8 assay was used to detect the viability of cells.The content of inflammatory factors was detected using ELISA assay.The expression of miRNA-432-5p mRNA in cells was detected by RT-qPCR.Hoechst staining was used to detect apoptosis.Expression of NLRP3,Caspase1,and IL-1β protein was detected by Western blot.Results OD value of cell viability in LPS group(6 h:0.809±0.09;12 h:0.601±0.09;24 h:0.485±0.43)were significantly lower than 1.000±0.00 in control group,with statistically significant differences(P<0.05).The mRNA expression level of miRNA-432-5p was 1.563±0.10 in LPS group,which was significantly higher than 1.000±0.00 in control group,and the apoptosis rate was(41.52±1.83)% in LPS group,which was significantly higher than(0.000±0.00)% in control group,with statistically significant differences(P<0.05).The content of TNF-α,IL-6,and IL-1β in culture medium were(36.32±6.07)pg/mL,(45.08±3.75)pg/mL,(40.87±6.65)pg/mL in LPS group,respectively,which were significantly higher than(5.27±0.84)pg/mL,(5.57±0.65)pg/mL,(3.26±0.85)pg/mL in the control group,with statistically significant differences(P<0.05).The expression levels of NLRP3,Caspase1,and IL-1β protein were 0.734±0.08,0.305±0.06,0.430±0.05 in LPS group,respectively,which were significantly higher than 0.163±0.03,0.025±0.01,0.032±0.01 in the control group,with statistically significant differences(P<0.05).In miRNA-432-5p mimics group,the apoptosis rate was(24.23±3.09)%,which was significantly lower than(41.52±1.83)% in LPS group,and the cell viability was 0.639±0.09,which was significantly higher than 0.468±0.07 in LPS group,with statistically significant differences(P<0.05).The content of TNF-α,IL-6,and IL-1β in culture medium in miRNA-432-5p mimics g

关 键 词：miRNA-432-5p NOD样受体热蛋白结构域相关蛋白3 凋亡 细胞损伤 

分 类 号：R563[医药卫生—呼吸系统]