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. 2021 Aug 23;297(3):101111. doi: 10.1016/j.jbc.2021.101111

Figure 7.

Figure 7

PI3K/AKT activation was involved in ghrelin-mediated protection of lung injury in vivo.A, immunofluorescence staining of p-AKT showed that increased pAkt in the ghrelin treated but not the LPS-only treated lungs. Red (TRICT) represents p-AKT, and blue represents nuclei. Mice divided into four groups were pretreated with or without ghrelin and subjected to intranasal injection with LPS (3 mg/kg) or dimethyl sulfoxide (DMSO), including the control group, LPS group, ghrelin group, and LPS + ghrelin group. B, the absorbance value of p-AKT in cells. Three slices of each mouse were used, and the observer randomly observed and photographed four fields of view. The observers were blinded to each experimental group. Data are expressed as the mean ± SEM, n = 4, ##p < 0.05 compared with the LPS group, ∗p < 0.05 compared with the control group, #p < 0.05 compared with the LPS group. C, H&E staining (magnification, ×200) showed that LY294002 aggravated histological injury in LPS-induced ARDS mice (n = 6 independent mice from each group analyzed in triplicate). Mice divided into four groups were pretreated with or without ghrelin and subjected to intranasal injection with LPS (3 mg/kg) or DMSO, including the control group, LPS group, ghrelin + LPS group, and ghrelin + LY294002 + LPS group. PI3K inhibitor, LY294002, dissolved in DMSO was injected into mice 1 h before LPS or PBS instillation through the tail vein. D, lung injury scores were utilized for quantitative analysis of lung histopathologic damage. E, immunofluorescence staining of VE-cadherin showed that ghrelin reversed the LPS-induced decrease in the expression of VE-cadherin, but this effect disappeared after adding LY294002. Green (TRICT) represents VE-cadherin, and blue represents nuclei. Mice divided into five groups were pretreated with or without ghrelin and subjected to intranasal injection with LPS (3 mg/kg) or DMSO, including the control group, ghrelin group, LPS group, ghrelin + LPS group, and ghrelin + LY294002 + LPS group. F, the absorbance value of VE-cadherin in cells. Three slices of each mouse were used, and the observer randomly observed and photographed six fields of view. The observers were blinded to each experimental group. Data are expressed as the mean ± SEM, n = 6, ∗p < 0.05 compared with the control group, #p < 0.05 compared with the LPS group, ∗∗p < 0.05 compared with the ghrelin + LPS group. ARDS, acute respiratory distress syndrome; LPS, lipopolysaccharide; VE-cadherin, vascular endothelial cadherin.