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Objective:To evaluate the effects of bosutinib on acute lung injury in mice with endotoxemia.Methods:Sixty clean-grade healthy male C57BL/6 mice, aged 8-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=15 each) using a random number table method: control group (group C), bosutinib group (group B), endotoxemia group (group lipopolysaccharide [LPS]) and bosutinib plus endotoxemia group (group B+ LPS). Septic acute lung injury model was developed by intraperitoneal injection of LPS.Bosutinib 5 mg/kg was injected via the tail vein at 0.5 h before establishing the model in group B+ LPS and at the corresponding time point in group B. At 24 h after developing the model, the mice were sacrificed for microscopic examination of the pathological results of lung tissues which were scored for calculation of the lung coefficient (LI) and wet/dry lung weight (W/D) ratio, and for determination of the content of Evans blue in lung tissues (by Evans blue staining), expression of vascular endothelial cadherin (VE-cadherin), vascular cell adhesion molecule 1 (VCAM-1), phosphorylated nuclear transcription factor κB p65 (p-NF-κB p65), phosphorylated nuclear factor κB inhibitory protein α (pIκB-α) (by Western blot) and expression of interleukin-1beta (IL-1β), IL-6 and tumor necrosis factor-alpha (TNF-α) mRNA (using real-time polymerase chain reaction). Results:Compared with group C, the LI, W/D ratio, Evans blue content in lung tissues and lung injury score were significantly increased, and the expression of IL-1β mRNA, TNF-α mRNA, IL-6 mRNA, VCAM-1, p-NF-κB p65 and pIKB-α was up-regulated, and the expression of VE-cadherin was down-regulated in group LPS ( P<0.05), and no significant change was found in the parameters mentioned above in group B ( P>0.05). Compared with group B, the LI, W/D ratio, Evans blue content in lung tissues and lung injury score were significantly decreased, and the expression of IL-1β mRNA, TNF-α mRNA, IL-6 mRNA, VCAM-1, p-NF-κB p65 and pIKB-α was down-regulated, and the expression of VE-cadherin was up-regulated in group LPS ( P<0.05). Conclusions:Bosutinib can ameliorate the acute lung injury in mice with endotoxemia.
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Objective To explore the effect of bosutinib on cell differentiation and apoptosis of human acute myeloid leukemia (AML) cell lines including HL-60, THP-1, and U937, and to clarify the related mechanisms. Methods MTT Assay was used to assess the proliferation of HL-60, THP-1, and U937 cells treated with various concentrations of bosutinib (0-20 μmol/L) for 48 h. Cell surface marker CD11b expression was detected by flow cytometry in HL-60, THP-1, and U937 cells treated with bosutinib (0-5 μmol/L) for 72 h. Apoptosis was evaluated by Annexin V-FITC/PI double stainning in the following two sets of experiments: 1 HL- 60, THP-1 and U937 cells were treated with various concentrations of bosutinib (0-10 μmol/L). 2 U937 cells were pretreated with caspases inhibitor benzyloxy-carbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK) for 1 h and then exposed to bosutinib for 48 h. Western blot was used to examine the protein expression of differentiation related transcription factors C/EBPβ, P21, and c-Myc, and apoptosis related protein Mcl- 1, Bax, and Caspase 3 in HL- 60 and U937 cells treated with bosutinib for 24 h. Results Bosutinib dose- dependently inhibited AML cell growth. Treatment with various concentrations of bosutinib for 72 h significantly increased CD11b expression and apoptotic rate in AML cells as compared to blank control (P< 0.05 or P < 0.01). Bosutinib effectively upregulated protein expression levels of C/EBPβ and P21 in HL-60 cells, and induced down-regulation of Mcl-1 with up-regulation of Bax protein expression, and activated Caspase 3 in U937 ells. Pretreatment of U937 cells with Z-VAD-FMK significantly inhibited apoptosis induced by bosutinib. Conclusion Bosutinib effectively inhibits cell proliferation and induces cell differentiation with apoptosis in AML cells. It could be a potent therapeutic agent against AML.