The Function And Mechanism Of LncRNA-loc105377478 In Inflammation Of Human Bronchial Epithelial Cells Induced By Nano-neodymium Oxide

Objective To explore the changes of lnc RNA loc105377478 in the inflammatory response induced by rare earth nano-neodymium oxide in human bronchial epithelial cells(16HBE),and investigate its function and mechanism of action.Method ELISA and q RT-PCR were used to detect the expression levels of IL-8 and IL-6 in 16 HBE cells that exposed to different doses of neodymium oxide for 24、48and72 hours,CCK8 kit was used to detect cell viability,ROS kit was used to detect reactive oxygen species in cells,LDH kit was used to detect cell membrane integrity.After differential expression of lnc RNA expression profile was detected by microarray,5 up-regulated and 5down-regulated genes were selected for verification.After Lnc RNA loc105377478 was ASOs/Over Expressed,the expressions of IL-6 and IL-8 were detected by ELISA and q RT-PCR,and the changes of ROS production and LDH release were also detected.After exposure to 16 HBE cells for 48 hours,the expression of ADIPOR1,p-IκB,and p-p65 proteins in the cells were detected by western blot for 48 hours.After Lnc RNA loc105377478 interference/overexpression,Western blot was used to detect the expression of p-IκB and p-p65 proteins in neodymium oxide-exposed 16 HBE cells and changes in target proteins predicted by lnc RNA loc105377478.Finally,the expression of IL-8 and IL-6 in 16 HBE cells exposed to nano-neodymium oxide was detected by ELISA using silencing and overexpression of lnc RNA loc105377478 combined with NF-κB pathway inhibitor.Results The study found that 10-80 μg/ml nano-neodymium oxide exposed to 16 HBE cells had no significant change in cell viability(p?0.05),120-320μg/ml nano-neodymium oxide exposed to 16 HBE cells had significant decreased in cell viabilityand.5-80μg/ml nano-neodymium oxide treatment of 16 HBE cells significantly reduced cell membrane integrity(p<0.01).The expression of inflammatory factors increased significantly(p<0.01).We performed chip(lnc RNA,m RNA)analysis on the infected samples,and selected 5 lnc RNAs that were significantly up-regulated and 5significantly down-regulated,respectively,and verified by q RT-PCR.p<0.01),and it may function by participating in multiple inflammatory pathways.Through functional experiments,it was found that the inflammation level of 16 HBE cells treated with nano neodymium oxide after interference with lnc RNA loc105377478 was significantly weakened(p<0.05).After overexpression of lnc RNA loc105377478,the inflammation level increased(p<0.05).Through bioinformatics analysis,ADIPOR1 may be involved in the regulation of NF-κB signaling pathway as an lnc RNA target protein.ADIPOR1 interference can lead to an increase in NF-κB pathway activating protein(p<0.05),and co-transfection with lnc RNA loc105377478 interference Changes in key proteins of the NF-κB pathway can be restored(p<0.05).Conclusions Nano neodymium oxide can cause inflammatory response in 16 HBE cells.The expression of lnc RNA loc105377478 was significantly up-regulated in 16 HBE cells induced by nano-neodymium oxide,and lnc RNA loc105377478 may promote the expression of IL-6 and IL-8 by regulating ADIPOR1/NF-κB.