摘要
目的研究lncRNA PCGEM1/miR-155-5p轴对LPS诱导的气管平滑肌细胞增殖和凋亡及炎症反应的影响。方法采用100μg/ml LPS刺激支气管平滑肌细胞BSMC 12 h以诱导细胞损伤。实时荧光定量PCR(RT-qPCR)检测细胞lncRNA PCGEM1和miR-155-5p表达水平;细胞计数试剂盒(CCK-8)法检测细胞增殖;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测细胞CyclinD1、Cleaved-caspase-3蛋白表达;双荧光素酶报告实验检测lncRNA PCGEM1和miR-155-5p靶向关系。结果与NC组比较,LPS组细胞lncRNA PCGEM1表达水平显著降低,miR-155-5p表达水平显著升高(P<0.05)。与pcDNA+LPS组比较,pcDNA-lncRNA PCGEM1+LPS组lncRNA PCGEM1表达水平显著升高,吸光度值、CyclinD1蛋白表达显著降低,细胞凋亡率、Cleaved-caspase-3蛋白表达显著升高,TNF-α、IL-6、IL-33水平显著降低(P<0.05)。与anti-miR-NC+LPS组比较,antimiR-155-5p+LPS组miR-155-5p表达水平显著降低,吸光度值、CyclinD1蛋白表达显著降低,细胞凋亡率、Cleaved-caspase-3蛋白表达显著升高,TNF-α、IL-6、IL-33水平显著降低(P<0.05)。与miR-NC+pcDNA-lncRNA PCGEM1+LPS组比较,miR-155-5p+pcDNA-lncRNA PCGEM1+LPS组细胞miR-155-5p表达水平显著升高,吸光度值、CyclinD1蛋白表达显著升高,细胞凋亡率、Cleaved-caspase-3蛋白表达显著降低,TNF-α、IL-6、IL-33水平显著升高(P<0.05)。结论lncRNA PCGEM1下调miR-155-5p降低LPS诱导的气管平滑肌细胞凋亡及炎症反应,抑制增殖。
This study was designed to evaluate the effect of lncRNA PCGEM1/miR-155-5p axis on LPSinduced proliferation,apoptosis and inflammation of bronchial smooth muscle cells(BSMCs).BSMCs were stimulated with 100μg/ml LPS for 12 h to induce cell dysfunction.Then RT-qPCR was used to detect the expression levels of lncRNA PCGEM1 and miR-155-5p;cell counting kit(CCK-8)assay and flow cytometry were used to detect cell proliferation and apoptosis;Western blot was used to detect the protein expression of CyclinD1 and Cleaved-caspase-3;Dual luciferase reporter assay was applied to detect the targeting relationship between lncRNA PCGEM1 and miR-155-5p.Thereafter,we found that the expression level of lncRNA PCGEM1 was decreased in the LPS-induced BSMCs,while miR-155-5p expression was significantly increased(P<0.05).Overexpression of lncRNA PCGEM1 inhibited the cell proliferation,the release of TNF-α,IL-6 and IL-33,induced the cell apoptosis,as well as reduced CyclinD1 protein and increased Cleaved-caspase-3 protein in LPS-stimulated BSMCs(P<0.05).Moreover,knockdown of miR-155-5p suppressed BSMC proliferation,inflammation,and induced cell apoptosis under LPS treatment(P<0.05).Importantly,miR-155-5p as a target of lncRNA RTPCGEM1,its overexpression attenuated the inhibitory effects of lncRNA PCGEM1 on the proliferation and inflammation,as well as the promoting effects on the apoptosis in LPS-stimulated BSMCs(P<0.05).Collectively,lncRNA PCGEM1 down-regulates miR-155-5p to reduce LPS-induced BSMC proliferation and inflammation,while inhibit BSMC apoptosis.
作者
唐瑞琪
皮光环
蔡艳
雷勋明
庞英
谢娜
刘韦
TANG Ruiqi;PI Guanghuan;CAI Yan;LEI Xunming;PANG Yin;XIE Na;LIU Wei(Department of Respirology and Hematology,Sichuan Provincial Maternity and Child Health Care Hospital,Chengdu 610041,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2023年第6期521-529,共9页
Immunological Journal
作者简介
通信作者:刘韦,E-mail:1191934704@qq.com。
