Research Paper Volume 15, Issue 24 pp 15640—15656

Figure 7. Fluoxetine up-regulates miR-450b-5p via Nrf2 signaling pathway in HK-2 Hypoxia/Reoxygenation (H/R) model. (A) mRNA expression level of miR-450b-5p after the transfection of miR-450b-5p over-expression vector in HK-2 cell line. (B) Protein expression level of Nrf2 after the transfection of miR-450b-5p over-expression vector in HK-2 cell line. (C) qRT-PCR analysis showed the expression level of miR-450b-5p from renal tissues in H/R group and H/R+FLX group after the injection of NC mimic, miR-450b-5p mimic or miR-450b-5p mimic+Recombinant protein Nrf2. (D) Cell viability of HK-2 in H/R group and H/R+FLX group after the injection of NC mimic, miR-450b-5p mimic or miR-450b-5p mimic+Recombinant protein Nrf2. (E) DHE staining of HK-2 cells in H/R group and H/R+FLX group after the injection of NC mimic, miR-450b-5p mimic or miR-450b-5p mimic+Recombinant protein Nrf2. ROS exhibit red fluorescence under fluorescent microscope (magnification ×400). (F) Immunohistochemical analysis showed the expression level of iNOS in H/R group and H/R+FLX group after the injection of NC mimic, miR-450b-5p mimic or miR-450b-5p mimic+Recombinant protein Nrf2 in HK-2 cell model (magnification ×400). (G) Western Blotting analysis showed the expression level of NF-κB, Nrf2 and Nrf2-dependent antioxidant enzymes, including HO-1, NQO1 and GST in H/R group and H/R+FLX group after the injection of NC mimic, miR-450b-5p mimic or miR-450b-5p mimic+Recombinant protein Nrf2 in HK-2 cell model. Data are expressed as mean ± SD. ^*significant difference vs. CON group (P < 0.05); ^#significant difference vs. H/R group (P < 0.05); ^&significant difference vs. H/R+FLX group (P < 0.05); ^@significant difference vs. H/R+FLX+miR-450b-5p mimic group (P < 0.05).