Affiliation:
1. Department of Geriatrics, The Affiliated Hospital of Southwest Medical University, Luzhou, China
2. Chengdu Medical College Southwest Medical University, Chengdu, China
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 (
). 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.
Subject
Complementary and alternative medicine
Cited by
1 articles.
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