RESUMO
Emerging evidence suggests that endothelial activation plays a central role in the pathogenesis of acute respiratory distress syndrome (ARDS) and multi-organ failure in patients with COVID-19. However, the molecular mechanisms underlying endothelial activation in COVID-19 patients remain unclear. In this study, the SARS-CoV-2 viral proteins that potently activate human endothelial cells were screened to elucidate the molecular mechanisms involved with endothelial activation. It was found that nucleocapsid protein (NP) of SARS-CoV-2 significantly activated human endothelial cells through TLR2/NF-{kappa}B and MAPK signaling pathways. Moreover, by screening a natural microbial compound library containing 154 natural compounds, simvastatin was identified as a potent inhibitor of NP-induced endothelial activation. Remarkablely, though the protein sequences of N proteins from coronaviruses are highly conserved, only NP from SARS-CoV-2 induced endothelial activation. The NPs from other coronaviruses such as SARS-CoV, MERS-CoV, HUB1-CoV and influenza virus H1N1 did not affect endothelial activation. These findings are well consistent with the results from clinical investigations showing broad endotheliitis and organ injury in severe COVID-19 patients. In conclusion, the study provides insights on SARS-CoV-2-induced vasculopathy and coagulopathy, and suggests that simvastatin, an FDA-approved lipid-lowering drug, may benefit to prevent the pathogenesis and improve the outcome of COVID-19 patients.
RESUMO
Objective@#To excavate the mechanism of the combination of Radix Ophiopogonis and Schisandra chinensis to treatatherosclerosisbased on network pharmacology to discuss its mechnism.@*Methods@#This paper excavated the associated proteins with Radix Ophiopogonis and Schisandra chinensis from the TCMGeneDIT database, and constructed the multicomponent protein network of Radix Ophiopogonis, Schisandra chinensis and proteins ApoE-/- mice were randomly divided into control group, model group, low, medium, high dose group and atorvastatin calcium group. Except the control group, other groups were fed with H10540 high fat diet for 12 weeks. From the 4th week, the atrovastatin calcium group was given atrovastatin calcium liquid 6 mg/kg by gavage. The low, medium and high dose groups were administed 4.68, 2.34 and 1.17 g/kg, respectively, once a day by gavage for 8 weeks. The oil red staining was applied to observe the pathological changes of atherosclerotic aortic wall. Western blot was subjected to detect the expression change of mitogen activated protein kinases p38 (p38), ATP binding cassette subfamily G member 1 (ABCG1), Toll like receptor 4 (TLR4), heat shock protein 90 alpha family class a member 1 (HSP90AA1), MMP-9 and arachidonate 5-lipoxygenase (ALOX5) in liver tissue, as well as nuclear factor related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) in brain tissue.@*Results@#It was found that eleven components were interacted with 37 proteins, forming a protein interaction network with 48 nodes and 190 boundaries without isolated nodes. Compared to the model group, the level of p-p38/p38 (2.12 ± 0.12, 1.76 ± 0.11, 1.69 ± 0.10 vs. 2.45 ± 0.16), TLR4 (1.98 ± 0.10, 1.64 ± 0.11, 1.55 ± 0.12 vs. 2.68 ± 0.06), HSP90AA1 (1.79 ± 0.10, 1.66 ± 0.09, 1.59 ± 0.11 vs. 2.06 ± 0.07), MMP9 (1.84 ± 0.11, 1.75 ± 0.12, 1.66 ± 0.08 vs. 2.68 ± 0.10) in liver tissue of low, medium and high dose groups significantly decreased, the level of ABCG1 (0.53 ± 0.08, 0.78 ± 0.09, 0.81 ± 0.10 vs. 0.45 ± 0.04), ALOX5 (0.59 ± 0.04, 0.67 ± 0.09, 0.88 ± 0.07 vs. 0.47 ± 0.02) in liver tissue of low, medium and high dose groups significantly increased (P<0.05). The expression of Nrf2 (1.62 ± 0.12, 1.32 ± 0.09, 1.14 ± 0.06 vs. 2.12 ± 0.08) in cytoplasm of brain tissue significantly decreased, and Nrf2 (1.12 ± 0.09, 1.61 ± 0.07, 1.68 ± 0.11 vs. 1.07 ± 0.08) in cell nucleus of brain tissue significantly increased. The expression of HO-1 (1.16 ± 0.09, 1.73 ± 0.11, 1.82 ± 0.08 vs. 1.05 ± 0.04) in brain tissue significantly increased.@*Conclusions@#Network pharmacology and molecular biology were used to elucidate the molecular mechanism of the combination of Schisandra chinensis and Ophiopogon japonicus in the prevention and treatment of atherosclerosis, also to validate the related mechanism via Nrf2 pathway, which provided a reference for the further study of the combined prescription.
RESUMO
Objective:To excavate the mechanism of the combination of Radix Ophiopogonis and Schisandra chinensis to treatatherosclerosisbased on network pharmacology to discuss its mechnism.Methods:This paper excavated the associated proteins with Radix Ophiopogonis and Schisandra chinensis from the TCMGeneDIT database, and constructed the multicomponent protein network of Radix Ophiopogonis, Schisandra chinensis and proteins ApoE-/- mice were randomly divided into control group, model group, low, medium, high dose group and atorvastatin calcium group. Except the control group, other groups were fed with H10540 high fat diet for 12 weeks. From the 4th week, the atrovastatin calcium group was given atrovastatin calcium liquid 6 mg/kg by gavage. The low, medium and high dose groups were administed 4.68, 2.34 and 1.17 g/kg, respectively, once a day by gavage for 8 weeks. The oil red staining was applied to observe the pathological changes of atherosclerotic aortic wall. Western blot was subjected to detect the expression change of mitogen activated protein kinases p38 (p38), ATP binding cassette subfamily G member 1 (ABCG1), Toll like receptor 4 (TLR4), heat shock protein 90 alpha family class a member 1 (HSP90AA1), MMP-9 and arachidonate 5-lipoxygenase (ALOX5) in liver tissue, as well as nuclear factor related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) in brain tissue.Results:It was found that eleven components were interacted with 37 proteins, forming a protein interaction network with 48 nodes and 190 boundaries without isolated nodes. Compared to the model group, the level of p-p38/p38 (2.12 ± 0.12, 1.76 ± 0.11, 1.69 ± 0.10 vs. 2.45 ± 0.16), TLR4 (1.98 ± 0.10, 1.64 ± 0.11, 1.55 ± 0.12 vs. 2.68 ± 0.06), HSP90AA1 (1.79 ± 0.10, 1.66 ± 0.09, 1.59 ± 0.11 vs. 2.06 ± 0.07), MMP9 (1.84 ± 0.11, 1.75 ± 0.12, 1.66 ± 0.08 vs. 2.68 ± 0.10) in liver tissue of low, medium and high dose groups significantly decreased, the level of ABCG1 (0.53 ± 0.08, 0.78 ± 0.09, 0.81 ± 0.10 vs. 0.45 ± 0.04), ALOX5 (0.59 ± 0.04, 0.67 ± 0.09, 0.88 ± 0.07 vs. 0.47 ± 0.02) in liver tissue of low, medium and high dose groups significantly increased ( P<0.05). The expression of Nrf2 (1.62 ± 0.12, 1.32 ± 0.09, 1.14 ± 0.06 vs. 2.12 ± 0.08) in cytoplasm of brain tissue significantly decreased, and Nrf2 (1.12 ± 0.09, 1.61 ± 0.07, 1.68 ± 0.11 vs. 1.07 ± 0.08) in cell nucleus of brain tissue significantly increased. The expression of HO-1 (1.16 ± 0.09, 1.73 ± 0.11, 1.82 ± 0.08 vs. 1.05 ± 0.04) in brain tissue significantly increased. Conclusions:Network pharmacology and molecular biology were used to elucidate the molecular mechanism of the combination of Schisandra chinensis and Ophiopogon japonicus in the prevention and treatment of atherosclerosis, also to validate the related mechanism via Nrf2 pathway, which provided a reference for the further study of the combined prescription.
