Role of macrophages in LPS-induced apoptosis of intestinal epithelial cells

doi:10.12092/j.issn.1009-2501.2019.10.009

Abstract

Abstract:

AIM: To evaluate the role and mechanism of macrophages on lipopolysaccharide (LPS)-induced apoptosis of intestinal epithelial cells. METHODS: NCM460 intestinal epithelial cells were incubated in regular culture medium (group R) or macrophage conditioned medium (group M) in 6-well plates. Different concentrations (0, 0.1, 10, 30 μg/mL) of LPS was added to the medium of NCM460 cells for 24 hours, then the cells were collected for Annexin V/7AAD staining, following with flow cytometry analysis. The cell culture supernatant was also collected to detect the TNF-α and IL-6 levels by enzyme-linked immunosorbent assay. Western blot was used to measure the activity of NF-κB signaling pathway. RESULTS:In the same culture medium, with the increase of LPS concentration, the levels of inflammatory factors TNF-α, IL-6 and apoptotic rate in the supernatant of cell culture medium increased gradually (P<0.05). Compared with group R, the percentage of apoptotic cells were significantly increased in group M when cells were treated with equal concentrations of LPS. Compared with NCM460 cells stimulated by LPS at the same dose, the apoptotic rate and the levels of inflammatory factors TNF-α and IL-6 in supernatant of M group were higher than those of R group (P<0.05). Compared with group R, the expression levels of IκB-α protein was down-regulated.CONCLUSION: Macrophages play an important role in LPS-induced apoptosis of intestinal epithelial cells. The mechanism may be related to the release of inflammatory cytokines by activation of NF-κB signaling pathway.

Key words: lipopolysaccharides, macrophages, apoptosis, enterocyte

CLC Number:

R965.2

CAO Yingya, CHEN Qun, WANG Zhen, WU Jingyi, JIANG Xiaogan, JIN Xiaoju, LU Weihua. Role of macrophages in LPS-induced apoptosis of intestinal epithelial cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(10): 1142-1146.

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