Dexmedetomidine Regulates the PI3K/AKT Signaling Pathway to Improve Lipopolysaccharide-induced Colitis Colon Epithelial Cell Injury
In order to study the effects of dexmedetomidine on inflammation,proliferation and apoptosis of human colon-ic epithelial NCM-460 cells induced by lipopolysaccharide(LPS)and the regulation of phosphatidylinositol-3-kinase/serine-threonine kinase(PI3K/AKT)signaling pathway,we cultured human colonic epithelial NCM-460 cells in vitro,they were di-vided into control group,LPS group(1 μg/mL LPS)and dexmedetomidine groups with different mass concentrations(1 μg/mL LPS+1.25,2.50,5.00 and 10.00 μg/mL dexmedetomidine).After 24 hours'intervention,the appropriate mass concentration of dexmedetomidine was screened out for subsequent experiments.Then,The human colonic epithelial NCM-460 cells were di-vided into control group,LPS group,dexmedetomidine group(1 μg/mL LPS+5.00 μg/mL dexmedetomidine),LY294002 group(1 μg/mL LPS+10 μmol/L PI3K/AKT pathway inhibitor LY294002)and inhibitor group(1 μg/mL LPS+5.00 μg/mL dexme-detomidine+10 μmol/L LY294002),and activator group(1 μg/mL LPS+5.00 μg/mL dexmedetomidine+10 μmol/L PI3K/AKT pathway agonist SC79)were intervened for 24 hours.Cell viability was detected by live cell counting kit-8(CCK-8).The ex-pression levels of tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)were detected by enzyme-linked immunosorbent assay(ELISA).The cell proliferation rate was determined by 5-ethynyl-2'deoxyuridine(EdU).The apoptosis rate was deter-mined by Hoechst 33258 staining.The expression levels of cyclin D1,cleaved cysteine protease 3(cleaved Caspase 3)and key proteins of PI3K/AKT signaling pathway in cells were determined by Western blot(WB).According to the cell viability and the expression level of inflammatory factor TNF-α,5.00 μg/mL dexmedetomidine was selected for the follow-up experiment.The results showed that compared with the control group,the cell proliferation rate and the expression level of Cyclin D1 in LPS group significantly decreased(P<0.05),while the cell apoptosis rate,the expression levels of TNF-α,IL-8,and cleaved Cas-pase 3,the ratio of p-PI3K/PI3K and p-AKT/AKT significantly increased(P<0.05).Dexmedetomidine and LY294002 in dex-medetomidine group and LY294002 group reversed the above-mentioned effects of LPS on human colonic epithelial NCM-460 cells(P<0.05).Compared with dexmedetomidine group,LY294002 in inhibitor group enhanced the effect of dexmedetomi-dine on human colonic epithelial NCM-460 cells induced by LPS(P<0.05),while SC79 in activator group weakened the effect of dexmedetomidine on human colonic epithelial NCM-460 cells induced by LPS(P<0.05).Studies have shown that dexme-detomidine can promote the proliferation of human colonic epithelial NCM-460 cells induced by LPS and inhibit its inflamma-tion and apoptosis,and its mechanism may be related to blocking the signal transduction of PI3K/PI3K signaling pathway.This study can provide a new direction for the treatment of colitis.
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