沉默lncRNA LINC00472对脂多糖诱导 肾小管上皮细胞损伤的影响及其机制.

Objective To investigate the effect and mechanism of silencing long non-coding RNA (lncRNA) LINC00472 on lipopolysaccharide (LPS)-induced renal tubular epithelial cell injury. Methods The human renal tubular epithelial cell line HK-2 (hereinafter referred to as HK-2 cells) was cultured in vitro and randomly divided into the control group, LPS group, LPS + si-LINC00472 group, LPS + si-NC group, LPS + miR-136-3p group, LPS + miR-NC group group, LPS + si-LINC00472 + anti-miR-136-3p group, and LPS + si-LINC00472 + anti-miR-NC group. Cells in the LPS + si-LINC00472 group were transfected with si-LINC00472, LPS + si-NC group with si-NC, LPS + miR-136-3p group with miR-136-3p, LPS + miR-NC group with miR-NC, LPS + si-LINC00472 + anti-miR- 136-3p group with si-LINC00472 + anti-miR-136-3p, and LPS + si-LINC00472 + anti-miR-NC group with si-LINC00472 + anti-miR-NC, followed by 1 μg/mL LPS stimulation for 24 h after 6-hour transfection, respectively. The cells in the LPS group were not transfected and were only stimulated with 1 μg/mL LPS for 24 h. The cells in the control group were routinely cultured without transfection and LPS stimulation. After the intervention, the cells in each group were collected, and RT-qPCR was used to detect the expression of lncRNA LINC00472 and miR-136-3p. The levels of IL-6 and IL-8 in the culture supernatant were detected by ELISA, and the binding sites of lncRNA LINC00472 and miR-136-3p, miR-136-3p and PDCD4 were predicted by Lnc-Base Predicted v.2 online prediction website and starBase database. The targeted regulatory relationship between lncRNA LINC00472 and miR-136-3p, miR-136-3p and PDCD4 was verified by dual-luciferase reporter gene experiments, and the protein expression of PDCD4 was detected by Western blotting. Results The relative expression of lncRNA LINC00472 in the LPS group was significantly higher than that in the control group, the relative expression of miR-136-3p was significantly lower than that in the control group, the cell viability was also significantly lower than that in the control group, and the levels of IL-6 and IL-8 in the culture supernatant were significantly higher than those of the control group (all P< 0. 05). The relative expression of lncRNA LINC00472 in the LPS + si-LINC00472 group was significantly lower than that in the LPS + si-NC group, the relative expression of miR-136-3p was significantly higher than that in the LPS + si-NC group, the cell activity was also significantly higher than that in the LPS + si-NC group, and the levels of IL-6 and IL-8 in the culture supernatant were significantly lower than those in the control group (all P<0. 05). The relative expression level of miR-136-3p and cell activity in LPS + miR-136-3p group were significantly higher than those in the LPS + miR-NC group, while the apoptosis rate, the levels of IL-6 and IL-8 in the culture supernatant and the relative expression of PDCD4 protein were significantly lower than those in the LPS + miR-NC group (all P<0. 05). The relative expression level of miR- 136-3p and the cell activity in the LPS + si-LINC00472 + anti-miR-136-3p group were significantly lower than those in LPS + si-LINC00472 + anti-miR-NC group, but the levels of IL-6 and IL-8 in culture supernatant and the relative expression of PDCD4 protein were significantly higher than those in the LPS + si-LINC00472 + anti-miR-NC group (all P<0. 05). Predicted by LncBase Predicted v.2 online prediction website and starBase database and verified by dual luciferase reporter gene experiments, lncRNA LINC00472 had a binding site with miR-136-3p, and miR-136-3p had a binding site with PDCD4. Conclusion Silencing lncRNA LINC00472 could alleviate LPS-induced renal tubular epithelial cell injury by promoting the expression of miR-136-3p and inhibiting the expression of PDCD4. [ABSTRACT FROM AUTHOR]