XIST Inhibition Attenuates Calcium Oxalate Nephrocalcinosis-Induced Renal Inflammation and Oxidative Injury via the miR-223/NLRP3 Pathway

Figure 2

Inhibition of XIST attenuated CaOx-induced renal inflammation and oxidative injury in vitro. COM at various doses (0 μg/ml, 50 μg/ml, 100 μg/ml, 250 μg/ml, and 500 μg/ml) was cocultured with HK-2 cells, and the cell viability was detected. (b) Cells were cocultured with 100 μg/ml COM for different amounts of time (0 h, 6 h, 12 h, 24 h, and 48 h), and the cell viability was examined. (c, d) qRT-PCR was used to detect the expression of XIST, NLRP3, Caspase-1, and IL-1β in HK-2 cells. (e, f) Western blotting also showed changes in the protein levels of NLRP3, Caspase-1, and IL-1β in HK-2 cells. (g) The generation of SOD, LDH, MDA, and H₂O₂ after different treatments was examined. (h) Flow cytometry was performed to assess cell necrosis. (i) TMRE was used to determine the mitochondrial membrane potential. Flow cytometry (j) and DCFH-DA (k) were used to detect ROS production in fluorescently labeled HK-2 cells. The data are presented as the of three independent experiments. ; , versus the NC group, #, versus the COM+si-NC group, as determined by one-way ANOVA (a–d, f–k).

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