H19 Promote Calcium Oxalate Nephrocalcinosis-Induced Renal Tubular Epithelial Cell Injury via a CeRNA Pathway

Abstract

Background: Intrarenal calcium oxalate (CaOx) crystals trigger inflammation and induce kidney tubular cell injury, which are processes that involve TLR4/NF-κB signalling. A recent genome-wide gene expression profile analysis of Randall's plaques in CaOx stone patients revealed that the expression of the long noncoding RNA H19 was significantly upregulated. However, to date, its role in kidney CaOx stones has not been reported. Method: GEO dataset were utilized to analyze gene expression profiles. Luciferase reporter assays, western blotting, qRT-PCR assay, immunoflfluorescence staining and ROS assay were employed to study the molecular mechanism of regulation of HMGB1/TLR4/NF-κB by H19 and miR-216b. In vitro and in vivo assays were performed to further confirm the the proinflammatory and prooxidative stress effects. Finding: H19 expression was significantly increased and positively corelated with the expression levels of HMGB1, TLR4 and NF-κB in the Randall's plaques and glyoxylate-induced CaOx nephrocalcinosis mouse model. H19 interacted with miR-216b and suppressed its expression. Additionally, miR-216b inhibited HMGB1 expression by directly binding its 3'-untranslated region (3'UTR). Moreover, the downregulation of H19 inhibited HMGB1, TLR4 and NF-κB expression and suppressed CaOx nephrocalcinosis-induced renal tubular epithelial cell injury, NADPH oxidase, and oxidative stress in vivo and in vitro. Interestingly, the inhibition of miR-216b can partially reverse the inhibitory effect of H19 knockdown on HMGB1 expression. Interpretation: We determined that H19 might serve as a facilitatory factor in the process of CaOx nephrocalcinosis-induced oxidative stress and renal tubular epithelial cell injury and revealed the mechanism by which the interaction between H19 and miR-216b could exert its effect via the HMGB1/TLR4/NF-κB pathway. Funding: This work was supported by the National Nature Science Foundation of China (8196030190, 8190033175, 81370805, 81470935, 81500534, and 81602236). Declaration of Interest: The authors declare no conflicts of interest. Ethical Approval: All animal experiments were performed according to protocols approved by the Ethics Committee of Tongji Hospital, Huazhong University of Science and Technology.