Inflammatory effects of high-glucose, high-free fatty acids, and heparin binding protein on HK-2neutrophil co-culture system
- Author:
WU Hong
;
WANG Ningrui
;
ZOU Haihong
- Publication Type:Journal Article
- Keywords:
High-glucose and high-free fatty acids;
heparin binding protein;
HK-2 cells;
inflammatory effects
- From:
China Tropical Medicine
2024;24(9):1100-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of high glucose, high free fatty acids (FFA), and heparin binding protein (HBP) treatment on the inflammatory response, oxidative stress, and cell apoptosis of the co-culture system established by human renal cortex proximal tubule epithelial cells (HK-2)/neutrophil cells. Methods Neutrophils were isolated from the peripheral blood of healthy individuals. Both HK-2 cells and neutrophil suspensions were mixed at a concentration of 105/mL to establish a co-cultured cell system. The treatment concentrations of lipopolysaccharide (LPS) and sodium heparin (Hep) were determined through the CCK-8 experiment. The concentrations of heparin binding protein (HBP), superoxide dismutase (SOD), inflammatory cytokines (IL-1β, IL-6, and IL-8), and the percentage of cells apoptosis was detected by quantitative immunofluorescence method, pyrogallol autoxidation method, the method based on the cytometric bead array (CBA) technology, and the detecting the expression of Annexin V-FITC fluorescence by flow cytometry, respectively. Results The treatment concentrations of LPS and Hep were determined respectively as 10 μg/mL and 50 μmol/L using the CCK-8 experiment. Compared with the normal group, treatment with high glucose and high FFA significantly increased the levels of HBP (4.11±0.07), SOD (2.67±0.06), IL-1β (11.28±0.35), IL-6 (2.57±0.10), IL-8 (1.32±0.05), and the incidence of cells apoptosis (1.08±0.04). LPS treatment (stimulation of HBP production) further exacerbated the relative levels of HBP (7.09±0.08), SOD (3.07±0.14), and IL-1β (22.96±0.49), IL-6 (2.79±0.11), IL-8 (1.55±0.07), and the incidence of cells apoptosis (1.17±0.05). Hep treatment (reducing HBP production) significantly reversed the effects induced by LPS on HBP (5.43±0.09), SOD (2.76±0.07), IL-1β (16.51±0.56), IL-6 (2.64±0.16), IL-8 (1.47±0.06), and the incidence of cells apoptosis (1.13±0.05). Conclusions Both the status of high glucose combined with high FFA, as well as HBP can affect the function of renal tubular epithelial cells. The underlying mechanisms may be related to promoting oxidative stress and inflammatory response.
- Full text:202511111545285733014.Inflammatory effects of high-glucose, high-free fatty acids, and heparin binding protein on HK-2neutrophil co-culture system.pdf
