白果内酯调控LEF1-AS1/miR-23b-3p分子轴缓解肺炎链球菌诱导的肺泡上皮细胞损伤  

Bilobalide regulates LEF1-AS1/miR-23b-3p molecular axis to alleviate alveolar epithelial cell injury induced by Streptococcus pneumoniae

作　　者：金晓立[1] 许建平[1] 黄曦[2] 曾云祥[1] JIN Xiaoli;XU Jianping;HUANG Xi;ZENG Yunxiang(Department of Clinical Laboratory,Wenzhou Hospital of Traditional Chinese Medicine,Wenzhou 325000,China;Department of Chinese Pharmacy,Wenzhou Hospital of Traditional Chinese Medicine,Wenzhou 325000,China)

机构地区：[1]温州市中医院检验科,325000 [2]温州市中医院中药房,325000

出　　处：《免疫学杂志》2022年第1期15-21,共7页Immunological Journal

基　　金：温州市科技项目(Y20160500)。

摘　　要：目的探讨白果内酯对肺炎链球菌诱导的肺泡上皮细胞损伤的保护作用及其对lncRNA LEF1-AS1/miR-23b-3p分子轴的调控作用。方法用肺炎链球菌诱导肺泡上皮细胞建立细胞损伤模型(感染组);分别转染si-NC、si-LEF1-AS1后,用肺炎链球菌感染细胞(感染+si-NC组和感染+si-LEF1-AS1组);pcDNA、pcDNA-LEF1-AS1分别转染至肺泡上皮细胞后用白果内酯与肺炎链球菌共同处理细胞(感染+白果内酯+pcDNA组和感染+白果内酯+pcDNA-LEF1-AS1组)。ELISA法检测TNF-α、IL-1β、IL-6的水平;流式细胞术检测细胞凋亡率;qRT-PCR法检测LEF1-AS1、miR-23b-3p的表达量;双荧光素酶报告基因实验检测LEF1-AS1与miR-23b-3p的靶向关系;Western blot检测Bcl-2、Bax蛋白表达量。结果白果内酯能够降低肺炎链球菌诱导的肺泡上皮细胞中TNF-α、IL-1β、IL-6的水平和LEF1-AS1的表达量(P<0.05),并可降低凋亡率和Bax蛋白水平(P<0.05),还可促进miR-23b-3p和Bcl-2表达(P<0.05),且呈剂量依赖性;LEF1-AS1可靶向调控miR-23b-3p;转染si-LEF1-AS1可降低TNF-α、IL-1β、IL-6的水平以及凋亡率和Bax蛋白水平(P<0.05),增加Bcl-2蛋白水平(P<0.05);转染pcDNA-LEF1-AS1能够逆转白果内酯对肺炎链球菌诱导的肺泡上皮细胞炎性因子及凋亡的作用。结论白果内酯可通过调控LEF1-AS1/miR-23b-3p分子轴抑制肺炎链球菌诱导的肺泡上皮细胞炎性反应及凋亡,进而减轻细胞损伤。To explore the protective effect of bilobalide on alveolar epithelial cells from Streptococcus pneumoniae-induced injury and its regulatory effect on the molecular axis of lncRNA LEF1-AS1/miR-23b-3p.Streptococcus pneumoniae was used to induce alveolar epithelial cells for establishing a cell injury model(infected group);alveolar epithelial cells were transfected with si-NC or si-LEF1-AS1,and then infected with Streptococcus pneumoniae(infection+si-NC group or infection+si-FEF1-AS1 group);pcDNA and pcDNA-LEF1-AS1 were respectively transfected into alveolar epithelial cells and treated with bilobalide and Streptococcus pneumoniae(infection+bilobalide+pcDNA).ELISA method was used to detect the levels of TNF-α,IL-1βand IL-6;flow cytometry was used to detect the apoptosis rate;qRT-PCR method was used to detect the expression of LEF1-AS1 and miR-23b-3p;the dual luciferase reporter gene experiment was carried out to detect the targeting relationship between LEF1-AS1 and miR-23b-3p;while Western blot was used to detect the expression of Bcl-2 and Bax proteins.Data showed that bilobalide could reduce the levels of TNF-α,IL-1βand IL-6,down-regulate the expression of LEF1-AS1 in alveolar epithelial cells induced by Streptococcus pneumoniae(P<0.05),and could reduce the rate of apoptosis and the protein level of Bax(P<0.05),but promote the expression of miR-23b-3p and Bcl-2(P<0.05)in a dose-dependent manner.LEF1-AS1 could target and regulate miR-23b-3p.Transfection of siLEF1-AS1 could reduce the levels of TNF-α,IL-1β,IL-6,the rate of apoptosis and the protein level of Bax(P<0.05),while increae the protein level of Bcl-2(P<0.05);transfection of pcDNA-LEF1-AS1 could reverse the effect of bilobalide on the inflammatory factors and apoptosis of alveolar epithelial cells induced by Streptococcus pneumoniae.Takn together,bilobalide could inhibit the inflammatory response and apoptosis of Streptococcus pneumoniae-induced alveolar epithelial cells by regulating the molecular axis of LEF1-AS1/miR-23b-3p,thereby alleviating cell

关 键 词：白果内酯 lncRNA LEF1-AS1 miR-23b-3p 肺泡上皮细胞 凋亡 

分 类 号：R378.1[医药卫生—病原生物学]