Xiangyong Kong1,2, Mei Yang1,2, Jing Guo1,2, Zhichun Feng1,2,3,4. 1. Newborn Care Center, Bayi Children's Hospital, The Seventh Medical Center, General Hospital of the People's Liberation Army. 2. Clinical Medical College, The Seventh Medical Center, General Hospital of the People's Liberation Army, Southern Medical University. 3. Beijing Key Laboratory of Pediatric Organ Failure. 4. National Engineering Laboratory for Birth defects prevention and control of key technology, Beijing, China.
Abstract
OBJECTIVE: To determine the effects of bovine lactoferrin (bLF) on cell viability, proliferation, and the protective roles in intestinal epithelial cells-6 (IEC-6) treated by lipopolysaccharide (LPS). METHODS: Cell viability and proliferation of IEC-6 were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Brdu assay separately. Cell cycle distribution was analyzed by flow cytometry. Inflammatory cytokines were analysed by real-time PCR and ELISA. Western blot was utilized to measure the level of MAPK and NF-κβ nuclear translocation. RESULTS: Dose-dependent effects of bLF on cell viability and proliferation were observed in IEC-6 cells (both P < 0.05), especially at a dose of 100 μg/ml. The percentage of cells in the G2 and S phase was significantly higher than those of the control group (8.17 ± 0.49% vs 4.72 ± 0.55%, P < 0.01 and 12.75 ± 0.33% vs 9.48 ± 0.33%, P < 0.01, respectively). The mRNA level of IL-1β, IL-6 and TNF-α was decreased by co-stimulation of bLF and LPS compared with the LPS treatments alone in IEC-6 cells (all P < 0.001). The secretion of IL-6 and TNF-α were also decreased by co-stimulation of bLF and LPS (both P < 0.01). Bovine lactoferrin treatment at dose of 100 μg/ml could inhibit the activation of MAPK/NF-κβ signal pathway induced by LPS (both P < 0.001). CONCLUSIONS: Bovine lactoferrin could promote the cell viability and proliferation, and have anti-inflammatory effects via inhibition of the activation of MAPK and NF-κβ nuclear translocation. Supplementation of formula with bLF may be beneficial in preventing NEC in preterm infants.
OBJECTIVE: To determine the effects of bovinelactoferrin (bLF) on cell viability, proliferation, and the protective roles in intestinal epithelial cells-6 (IEC-6) treated by lipopolysaccharide (LPS). METHODS: Cell viability and proliferation of IEC-6 were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Brdu assay separately. Cell cycle distribution was analyzed by flow cytometry. Inflammatory cytokines were analysed by real-time PCR and ELISA. Western blot was utilized to measure the level of MAPK and NF-κβ nuclear translocation. RESULTS: Dose-dependent effects of bLF on cell viability and proliferation were observed in IEC-6 cells (both P < 0.05), especially at a dose of 100 μg/ml. The percentage of cells in the G2 and S phase was significantly higher than those of the control group (8.17 ± 0.49% vs 4.72 ± 0.55%, P < 0.01 and 12.75 ± 0.33% vs 9.48 ± 0.33%, P < 0.01, respectively). The mRNA level of IL-1β, IL-6 and TNF-α was decreased by co-stimulation of bLF and LPS compared with the LPS treatments alone in IEC-6 cells (all P < 0.001). The secretion of IL-6 and TNF-α were also decreased by co-stimulation of bLF and LPS (both P < 0.01). Bovinelactoferrin treatment at dose of 100 μg/ml could inhibit the activation of MAPK/NF-κβ signal pathway induced by LPS (both P < 0.001). CONCLUSIONS:Bovinelactoferrin could promote the cell viability and proliferation, and have anti-inflammatory effects via inhibition of the activation of MAPK and NF-κβ nuclear translocation. Supplementation of formula with bLF may be beneficial in preventing NEC in preterm infants.