RESUMO
ObjectiveTo explore the effect of Ganoderma leucocontextum ethanol extract (GLE) on silicosis and its potential molecular mechanism using network pharmacology, molecular docking technology and animal experiments. Methods i) The components of GLE were analyzed using ultra-performance liquid chromatography-Q Exactive-mass spectrometry (UPLC-QE-MS) method. The active components, potential molecular pathways and targets of GLE in the intervention of inflammation process of silicosis was explored using network pharmacology and molecular docking technology. ii) Specific pathogen free male C57BL6/J mice were divided into four groups with 10 mice in each group. The mice in the silicosis model group and GLE intervention group were given a dose of 80 μL silica suspension with a mass concentration of 50 g/L once by non-exposed tracheal instillation, and the mice in the blank control group and GLE control group were given an equal volume of sterile 0.9% sodium chloride solution. From the second day after modeling, GLE control group and GLE intervention group were given GLE at a dose of 200 mg/(kg•d) by gavage, while blank control group and silicosis model group were given the same volume of 0.9% sodium chloride solution by gavage, once per day for 35 days. After that, the histopathological changes of lung tissues of mice were observed, the lung mass coefficient, inflammation score and the ratio of collagen deposition area were calculated, and the levels of tumor necrosis factor (TNF) -α, interleukin (IL) -1β and IL-6 in the lung tissues of mice were detected by enzyme-linked immunosorbent assay. Results i) A total of 76 active components of GLE were detected by UPLC-QE-MS. Among them, 36 ingredients met the screening criteria of the five principles of drug-like components. A total of 67 potential targets of the 36 GLE active ingredients to improve the inflammatory response of silicosis were screened based on the network pharmacology theory. The result of Kyoto Encyclopedia of Genes and Genomes enrichment analysis and Gene Ontology functional analysis showed that IL signaling and cytokine signaling of immune cells played a key role in the process of anti-silicosis of GLE. The results of molecular docking showed that the top 10 targets based on the 67 intersection targets were TNF, IL6, B-cell lymphoma 2, cellular tumor antigen p53, Caspase-3 subunit p12, JUN, epidermal growth factor receptor, IL1B, 67 kDa matrix metalloproteinase-9 and prostaglandin G/H synthase 2. The result of protein-protein interaction analysis showed that glycyrrhetinic acid had the strongest affinity with the key targets TNF-α, IL-1β and IL-6, followed by ganoderma acid DM, alismatol C, ganoderma acid β and red sapogenin. ii) The results of histopathological examination showed that the inflammatory response and collagen deposition were alleviated in the lungs of mice with silicosis. The lung mass coefficient, inflammation score, ratio of collagen deposition area and IL-6 expression in lung were lower in mice of the GLE intervention group (all P<0.05), compared with the silicosis model group. However, there was no significant difference in the levels of TNF-α and IL-1β in lung tissues between the two groups (all P>0.05). Conclusion GLE may reduce silica-induced lung inflammation and fibrosis by inhibiting the IL-6 level in lung tissues of mice. Its mechanism is associated with the synergistic action of multi-components, multi-targets and multi-pathways.
RESUMO
Angiokinases, such as vascular endothelial-, fibroblast- and platelet-derived growth factor receptors (VEGFRs, FGFRs and PDGFRs) play crucial roles in tumor angiogenesis. Anti-angiogenesis therapy using multi-angiokinase inhibitor has achieved great success in recent years. In this study, we presented the design, synthesis, target identification, molecular mechanism, pharmacodynamics (PD) and pharmacokinetics (PK) research of a novel triple-angiokinase inhibitor WXFL-152. WXFL-152, identified from a series of 4-oxyquinoline derivatives based on a structure-activity relationship study, inhibited the proliferation of vascular endothelial cells (ECs) and pericytes by blocking the angiokinase signals VEGF/VEGFR2, FGF/FGFRs and PDGF/PDGFR simultaneously . Significant anticancer effects of WXFL-152 were confirmed in multiple preclinical tumor xenograft models, including a patient-derived tumor xenograft (PDX) model. Pharmacokinetic studies of WXFL-152 demonstrated high favourable bioavailability with single-dose and continuous multi-dose by oral administration in rats and beagles. In conclusion, WXFL-152, which is currently in phase Ib clinical trials, is a novel and effective triple-angiokinase inhibitor with clear PD and PK in tumor therapy.
RESUMO
OBJECTIVE: To investigate the effect of transforming growth factor β1( TGF-β1) type Ⅰ receptor kinase inhibitor SB431542 on epithelial-mesenchymal transition( EMT) induced by paraquat in type Ⅱ alveolar epithelial cells A549. METHODS: A549 cells were randomly divided into control group,paraquat group and TGF-β1 blockade group,with3 samples in each group. The cells in the control group were cultured without any treatment,cells in paraquat group were stimulated by paraquat of a final concentration of 20 μmol/L,cells in TGF-β1 blockade group were treated with paraquat with a final concentration of 20 μmol/L and SB431542 with a final concentration of 20 μg/L. After 5 days,cultured cells were harvested and observed for morphologic and phenotypic characteristics using inverted phase contrast microscope and scanning electron microscope. Cell migration was assayed by Transwell chamber. The expression of target protein was detected by Western blot. The enzyme-linked immunosorbent assay was used to detect the TGF-β1 levels. RESULTS: Under inverted phase contrast microscope and scanning electron microscope,A549 cells in control group grew normally,cells in paraquat group changed from epithelial morphology to mesenchymal morphology,cells in TGF-β1 blockade group reversed to epithelial cell morphology. The results of cell migration showed that the number of cells in the paraquat group passed through the membrane was higher than that in the control group and TGF-β1 blockade group( P < 0. 05). The relative expression of E-cadherin and zonula occluden-1 in paraquat group was decreased( P < 0. 05),while the relative expression of vimentin,α-smooth muscle actin,type I collagen,the phosphorylation Smad and Mad related protein( p-Smad) 2 and p-Smad3 was elevated( P < 0. 05),compared to control group and TGF-β1 blockade group. The levels of total TGF-β1 and active TGF-β1 increased in paraquat group than that in control group( P < 0. 05). CONCLUSION: Paraquat induced EMT in A549 cells by activating TGF-β1/Smads signaling pathway. Early treatment with SB431542 can inhibit paraquatinduced EMT by blocking TGF-β1/Smads signaling pathway.