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Protective effect and mechanisms of exogenous neutrophil gelatinase-associated lipocalin on lipopolysaccharide-induced injury of renal tubular epithelial cell

Author:
Li, Jing, Chen, Yana, Deng, Fuxing, Zhao, Shuangping
Source:
Biochemical and biophysical research communications 2019 v.515 no.1 pp. 104-111
ISSN:
0006-291X
Subject:
Lentivirus, Western blotting, autophagy, cell proliferation, cell viability, electron microscopy, epithelial cells, fluorescent antibody technique, gene targeting, lipopolysaccharides, neutrophils, protective effect, proteins
Abstract:
To investigate the protective effect of exogenous neutrophil gelatinase-associated lipocalin (NGAL) on the lipopolysaccharide-induced injury of renal tubular epithelial cells and its regulation of autophagy.Renal tubular epithelial cells were treated with lipopolysaccharide (LPS) at different concentrations (0–100 μg/mL) and at different times (0–24 h), the expression of NGAL was detected to determine the optimal time and concentration of LPS treatment. The NGAL gene knockdown lentivirus (NGAL-RNAi) was constructed and verified its knockdown rate and inhibition effect. Renal tubular epithelial cells were randomly divided into Control group, LPS group, LPS + NGAL group, NGAL-RNAi + LPS group, and NGAL-RNAi + LPS + NGAL group. Western blot and immunofluorescence tested the expression of autophagy-associated proteins, the changes in the number of autophagosomes were observed by electron microscopy, analyzed the role of exogenous NGAL.The study showed the expression of autophagy-associated proteins (LC3-II and Beclin-1) in NGAL-RNAi + LPS group was significantly lower than the LPS group (P < 0.0100). The expression of LC3-II and Beclin-1 in the NGAL-RNAi + LPS + NGAL group was significantly higher than the NGAL-RNAi + LPS group (P < 0.0100). After the addition of exogenous NGAL, the autophagosomes in the LPS + NGAL group and the NGAL-RNAi + LPS + NGAL group were significantly increased under the electron microscope compared with the LPS group and the NGAL-RNAi + LPS group, and the cell proliferation rate and cell viability was significantly higher than unjoined groups (P < 0.0500).NGAL knockdown can significantly reduce the level of autophagy and decrease the proliferation rate and viability of cells.The addition of exogenous NGAL can increase the level of autophagy. This suggests that NGAL may play a protective role in the LPS-induced injury of renal tubular epithelial cells by promoting autophagy.
Agid:
6448246