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Chinese Journal of Clinical Pharmacology and Therapeutics ›› 2019, Vol. 24 ›› Issue (10): 1142-1146.doi: 10.12092/j.issn.1009-2501.2019.10.009

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

CAO Yingya, CHEN Qun, WANG Zhen, WU Jingyi, JIANG Xiaogan, JIN Xiaoju, LU Weihua   

  1. Department of Anesthesiology and Critical Care Medicine, Yijishan Hospital, Wannan Medical College, Wuhu 241001, Anhui, China
  • Received:2018-08-19 Revised:2018-09-13 Online:2019-10-26 Published:2019-10-28

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

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