miR-543靶向HDAC3调控LPS诱导的气道上皮细胞损伤

miR-543 regulates LPS-induced airway epithelial cell injury through targeting HDAC3

目的:探讨miR-543靶向组蛋白脱乙酰酶3(HDAC3)对脂多糖(LPS)诱导的气道上皮细胞损伤的影响。方法:用50μg/ml的LPS诱导气道上皮细胞BEAS-2B,实时荧光定量PCR(RT-qPCR)和Western blot检测miR-543和HDAC3表达。流式细胞术检测细胞凋亡。ELISA试剂盒检测细胞培养液上清中IL-6、IL-1β、TNF-α含量。双荧光素酶报告实验和Western blot确定miR-543对HDAC3的靶向调控关系。将miR-543模拟物、HDAC3小干扰RNA分别转染BEAS-2B细胞,检测过表达miR-543或干扰HDAC3对LPS诱导的BEAS-2B细胞损伤的影响。结果:LPS处理后BEAS-2B细胞miR-543表达降低,HDAC3表达、细胞凋亡率升高,培养液上清中IL-6、IL-1β、TNF-α含量升高(P<0.05)。过表达miR-543或干扰HDAC3表达后,LPS诱导的BEAS-2B细胞凋亡率降低,培养液上清中IL-6、IL-1β、TNF-α含量降低(P<0.05)。过表达HDAC3能够逆转miR-543过表达对LPS诱导的BEAS-2B细胞中IL-6、IL-1β、TNF-α水平及凋亡的影响(P<0.05)。miR-543靶向负调控HDAC3表达。结论:miR-543通过靶向HDAC3能够抑制LPS诱导的气道上皮细胞凋亡和炎症反应。

Objective:To investigate the effect of miR-543 on lipopolysaccharide(LPS)-induced airway epithelial cell injury by targeting histone deacetylase 3(HDAC3).Methods:The airway epithelial cells BEAS-2B were induced with 50μg/ml LPS.Realtime quantitative PCR(RT-qPCR)and Western blot were used to detect miR-543 and HDAC3 expression.Flow cytometry was selected to detect apoptosis.ELISA kit was used to detect the levels of IL-6,IL-1β,TNF-αin the cell culture supernatant.The dual luciferase report experiment and Western blot were utilized to confirm the targeting regulation relationship between miR-543 and HDAC3.The miR-543 mimics and HDAC3 small interfering RNA were transfected into BEAS-2B cells,respectively.and the effects of overexpressing miR-543 or interfering with HDAC3 on LPS-induced BEAS-2B cell damage were detected.Results:After LPS treatment,the expression of miR-543 in BEAS-2B cells decreased,HDAC3 expression and apoptotic rate increased,and the levels of IL-6,IL-1β,TNF-αin the culture supernatant increased(P<0.05).After overexpressing miR-543 or interfering with HDAC3 expression,the apoptosis rate of BEAS-2B cells decreased,and the contents of IL-6,IL-1βand TNF-αin the culture supernatant decreased(P<0.05).Overexpression of HDAC3 could reverse the effect of miR-543 overexpression on the levels of IL-6,IL-1β,TNF-α,and apoptosis of BEAS-2B cells induced by LPS(P<0.05)miR-543 targets and negatively regulates HDAC3 expression.Conclusion:miR-543 can inhibit LPSinduced airway epithelial cell apoptosis and inflammatory response by targeting HDAC3.