摘要
为探讨环状RNA 0060745(circ_ular RNA 0060745,circ_0060745)靶向miR-194对LPS诱导的支气管上皮细胞16HBE炎症损伤的影响,用50μg/mL LPS刺激16HBE细胞24h,RT-qPCR检测circ_0060745和miR-194的相对表达量;FACS检测16HBE细胞的凋亡率;ELISA检测16HBE细胞上清液中IL-6和TNF-α的水平。分别转染circ_0060745小干扰RNA(small interfering RNA,siRNA)和miR-194模拟物至16HBE细胞,检测下调circ_0060745或过表达miR-194对LPS诱导的16HBE细胞炎症损伤的影响。双荧光素酶报告基因实验和RT-qPCR分析circ_0060745与miR-194的相互作用。结果显示,LPS处理后,16HBE细胞的凋亡率、IL-6和TNF-α表达量及circ_0060745表达量显著升高,miR-194表达量显著降低(均P<0.05)。下调circ_0060745表达或过表达miR-194均可减轻LPS诱导的16HBE细胞的凋亡程度和炎症损伤程度(均P<0.05)。circ_0060745可直接与miR-194结合,且circ_0060745负调控miR-194的表达(P<0.05)。下调miR-194表达可逆转下调circ_0060745表达对LPS诱导的16HBE细胞凋亡和炎症反应的影响(均P<0.05)。由此,下调circ_0060745可通过靶向上调miR-194表达减轻LPS诱导的支气管上皮细胞凋亡程度和炎症反应。
To investigate the effect of miR-194 targeted circular RNA 0060745(circ_0060745) on LPS-induced inflammatoryinjury of bronchial epithelial cell 16 HBE, 16 HBE cells were stimulated with 50 μg/mL LPS for 24 hours and RT-qPCR was used to analyze the expression of circ_0060745 and miR-194. FACS was applied to detect cell apoptosis rate and ELISA was used to measure the levels of IL-6 and TNF-α in cell supernatant. Circ060745 small interfering RNA(siRNA) and miR-194 mimics were transfected into 16 HBE cells, respectively, to manipulate the expression levels of circ_0060745 or miR-194. Dual luciferase reporter gene assay and RT-qPCR were used to analyze the interaction between circ_0060745 and miR-194. The results showed that the apoptosis rate, the levels of IL-6 and TNF-α and the expression of circ_0060745 in 16 HBE cells treated with LPS were all significantly increased(all with P<0.05), while the expression of miR-194 was significantly decreased(P<0.05). Down-regulation of circ_0060745 or over-expression of miR-194 alleviated LPS-induced apoptosis and inflammatory damage of 16 HBE cells(all with P<0.05). Circ060745 directly bounded to miR-194 and negatively regulated the expression of miR-194(P<0.05). In contrast, down-regulating miR-194 reversed the effect of circ_0060745 inhibition on LPS-induced apoptosis and inflammation of 16 HBE cells(all with P<0.05). In conclusion, down-regulating circ_0060745 alleviates LPS-induced apoptosis and inflammation of bronchial epithelial cells possibly by direct up-regulation of miR-194 expression.
作者
宋斌
葛宏
李宁
SONG Bin;GE Hong;LI Ning(Department of Respiratory Medicine ZiBo Central Hospital,Zibo 255020,China)
出处
《现代免疫学》
CAS
北大核心
2022年第6期499-504,515,共7页
Current Immunology