Abstract:Glutamine (Gln) can egulate expressions of several inflammatory cytokines while it is not clear if it can attenuate inflammatory injury induced by lipopolysaccharide (LPS). The sustentacular cells (SCs) of Bos taurus on their third generation were cultured with various concentrations (0.5, 1, 2, 4 and 8 mmol/L) of Gln for 12 h and the survival rates of the cells were measured using CCK-8 kit 4 h after changing medium. The cell survival rates were 90.81% treated with 2 mmol/L of Gln. The readily cultured SCs were randomly divided into 4 groups including control group (blank), LPS group (cocultured with 0.1 μg/mL LPS for 12 h), Gln group (cocultured with 2 mmol/L Gln for 12 h) and LPS+Gln group (treated with 0.1 μg/mL LPS for 4 h after cocultured with 2 mmol/L Gln for 12 h). Heat shock protein72 (HSP72), Interkeukin receptor-associated kinase-M (IRAK-M), Toll-interacting protein (Tollip) and Zinc finger protein (A20)mRNA and protein expressions were detected by using qRT-PCR and Western blot, respectively. Interleukin-1β (IL-1β), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-13 (IL-13) and Interleukin-17 (IL-17) protein expression were measured by using Enzyme linked immunosorbent assay (ELISA). The results showed that Gln could induce SCs HSP72, IRAK-M, Tollip and A20 expression. The HSP72, IRAK-M, Tollip and A20 levels were significantly (P<0.05) increased compared to blank control group while the above indicators were all significantly (P<0.05) decreased in LPS+Gln group compared with LPS group. Similarly, IL-1β, IL-6, IL-8, IL-10, IL-13 and IL-17 expressions were significantly (P<0.05) increased in LPS group compared to the blank control group while they were all significantly (P<0.05) decreased in LPS+Gln group compared with LPS group. The above results demonstrated that LPS can induce expressions of inflammatory cytokines and negative feedback regulatory factors including HSP72, IRAK-M, Tollip and A20. Gln can inhibite inflammatory cytokines induced by LPS and stimulate anti-inflammatory cytokines and thereby attenuates inflammatory injury.
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