Abstract
OBJECTIVE: To investigate the influence of KCNQ1OT1 on HK-2 apoptosis and inflammation in ARI and its molecular mechanism. METHODS: Normal cultivated HK-2 cells were used as negative control (NC) group. Three different concentrations of lipopolysaccharide (LPS) were used to treat the cells (5 μg/mL, 10 μg/mL, and 20 μg/mL). The groups included si-KCN1OT1+ LPS, si-NC + LPS, miR-30a-5p + LPS, pcDNA-NLRP3+si-KCNQ1OT1 + LPS group, miR-NC + LPS group, and pcDNA + si-KCNQ1OT1 + LPS group. CCK-8 and flow cytometry are used to measure cell viability and apoptosis, while RT-qPCR and Western blotting are used to detect KCNQ1OT1, miR-30a-5p, and NLRP3 mRNA. ELISA was used to detect the levels of TNF-α, IL-6, and IL-1β in HK-2 cells. The targeting relationship among KCNQ1OT1, miR-30a-5p, and NLRP3 was verified. RESULTS: After the intervention of LPS, the viability of HK-2 cells was decreased, while the apoptosis rates were increased. The mRNA and protein expressions of NLRP3 and KCNQ1OT1 were increased, while the mRNA and protein levels of miR-30a-5p were decreased (P < 0.05). The expressions of Bax and Cleaved-caspase-3 were downregulated after silencing KCNQ1OT1 and overexpressed miR-30a-5p. In addition, the viability of HK-2 cells was improved, and the apoptosis was reduced by inhibiting KCNQ1OT1 and overexpressed miR-30a-5p. Thus, KCNQ1OT1 modulated NLRP3 via targeting miR-30a-5p. Overexpression of NLRP3 reverses KCNQ1OT1 inhibition of LPS-induced apoptosis, activity, and inflammation in HK-2 cells. CONCLUSIONS: Through modulating the miR-30a-5p/NLRP3 axis, inhibition of KCNQ1OT1 may reduce HK-2 apoptosis and inflammation in LPS-induced ARI.