RESUMO
Based on network pharmacology, molecular docking, and in vitro experimental verification, this study aims to explore the effect of Albiziae Cortex-Tribuli Fructus combination on HSC-LX2 pyroptosis. Specifically, the targets of Albiziae Cortex, Tribuli Fructus, and hepatic fibrosis were retrieved from an online database and CNKI, and "drug-component-target" network and "drug-component-target-disease" network were constructed. Protein-protein interaction(PPI) network was established based on STRING. Metascape was employed for Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment, and the mechanism of Albiziae Cortex-Tribuli Fructus combination against liver fibrosis was predicted. Molecular docking was used to verify some of the results of network pharmacology, and in vitro experiment was carried out to further verify the above conclusions. According to the results of network pharmacological analysis, 25 active components and 439 targets of Albiziae Cortex-Tribuli Fructus combination and 152 anti-liver fibrosis targets were screened out, including nucleotide-binding oligomerization domain and leucine-rich-repeat-and pyrin-domain-containing 3(NLRP3) and caspase-1. The key targets were involved in 194 KEGG pathways in which the NOD-like receptor signaling pathway topped. The binding common targets were related to pyroptosis. The results of in vitro experiment showed that the pair-containing serum reduced the proliferation rate of HSC-LX2 and the content of reactive oxygen species(ROS), interleukin-18(IL-18), and interleukin-1ß(IL-1ß)(P<0.05). Western blot and qRT-PCR suggested that the protein and gene expression of NLRP3, caspase-1, α-smooth muscle actin(α-SMA), and gasdermin D(GSDMD) in HSC-LX2 increased after Angâ ¡ stimulation, and the expression decreased after the intervention of pair-containing serum(P<0.05). In summary, the pair-containing serum can inhibit the classic pathway of pyroptosis, which may be the anti-liver fibrosis mechanism. This is consistent with the predicted results of network pharmacology.
Assuntos
Medicamentos de Ervas Chinesas , Células Estreladas do Fígado , Humanos , Farmacologia em Rede , Simulação de Acoplamento Molecular , Proteína 3 que Contém Domínio de Pirina da Família NLR , Caspase 1/genética , Fibrose , Medicamentos de Ervas Chinesas/farmacologiaRESUMO
BACKGROUND: Current evidence suggests that liver fibrosis is reversible even at late stages. Pyroptosis is reportedly regulated by classical and non-classical pathways and is also involved in the activation of the human hepatic stellate cell line LX2. This study sought to identify regulatory pathways that pyroptosis of HSC during AngII-ROS-induced HSC activation and provides novel insights for anti-fibrosis therapy by targeting HSC. MATERIALS AND METHODS: All experiments were conducted in HSC-LX2. The expressions of α-SMA, NLRP3, Caspases-1, -4, -5, ASC and GSDMD-N were detected in HSC-LX2 cells induced with AngII by Western blot and qRT-PCR. CCK8 was used to detect cell proliferation and activity. 2'-7'dichlorofluorescin diacetate (DCFH-DA) was used to measure ROS generation. An LDH assay kit was used to detect LDH released from damaged cells, and ELISA was used to quantify IL-18 and IL-1ß levels. RESULTS: After AngII stimulation, HSC-LX2 cell viability, ROS, LDH, IL-18, and IL-1ß were increased compared with Control group. At the same time, the protein and mRNA levels of α-SMA, NLRP3, Caspases-1, -4, -5, ASC and GSDMD-N were increased. In addition, after NAC and NSA treatment, LDH, IL-18 and IL-1ß levels and the protein and mRNA expression of α-SMA, Caspases-4 and -5, and GSDMD-N were decreased. CONCLUSION: HSC-LX2 pyroptosis induced by AngII-ROS is mediated by the classical pathway involving NLRP3/Caspase-1 and the non-classical pathway involving Caspases-4 and -5. Our results provide compelling evidence that AngII could activate Caspases-4 and -5 by producing ROS.
Assuntos
Proteína 3 que Contém Domínio de Pirina da Família NLR , Piroptose , Angiotensina II/farmacologia , Caspase 1/metabolismo , Humanos , Inflamassomos/metabolismo , Interleucina-18/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , RNA Mensageiro , Espécies Reativas de Oxigênio/metabolismoRESUMO
BACKGROUND: Evidence suggests that cytoglobin (Cygb) may function as a tumor suppressor gene. METHODS: We immunohistochemically evaluated the expression of Cygb, phosphatidylinositol-3 kinase (PI-3K), phosphorylated (p)-Akt, Interleukin-6 (IL-6), tumor necrosis factor-α (TNFα) and vascular endothelial growth factor (VEGF) in 88 patients with 41 high-grade gliomas and 47 low-grade gliomas. Intratumoral microvessel density (IMD) was also determined and associated with clinicopathological factors. RESULTS: Low expression of Cygb was significantly associated with the higher histological grading and tumor recurrence. A significant negative correlation emerged between Cygb expression and PI3K, p-Akt, IL-6, TNFα or VEGF expression. Cygb expression was negatively correlated with IMD. There was a positive correlation between PI3K, p-Akt, IL-6, TNFα and VEGF expression with IMD.High histologic grade, tumor recurrence, decreased Cygb expression, increased PI3K expression, increased p-Akt expression and increased VEGF expression correlated with patients' overall survival in univariate analysis. However, only histological grading and Cygb expression exhibited a relationship with survival of patients as independent prognostic factors of glioma by multivariate analysis. CONCLUSIONS: Cygb loss may contribute to tumor recurrence and a worse prognosis in gliomas. Cygb may serve as an independent predictive factor for prognosis of glioma patients.
Assuntos
Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Glioma/metabolismo , Glioma/patologia , Globinas/metabolismo , Recidiva Local de Neoplasia/metabolismo , Adolescente , Adulto , Idoso , Análise de Variância , Citoglobina , Feminino , Humanos , Interleucina-1/metabolismo , Estimativa de Kaplan-Meier , Masculino , Microvasos/patologia , Pessoa de Meia-Idade , Análise Multivariada , Gradação de Tumores , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Modelos de Riscos Proporcionais , Proteínas Proto-Oncogênicas c-akt/metabolismo , Estudos Retrospectivos , Transdução de Sinais , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto JovemRESUMO
At different temperatures, the binding of naphthol green B (NGB) to bovine serum albumin (BSA) was studied by the fluorescence spectroscopy, three-dimensional fluorescence spectrum, synchronous fluorescence spectrum and ultra-violet spectrum. It was shown that this compound has a quite strong ability to quench the fluorescence from BSA. After analyzing the fluorescence quenching data according to Sterm-Volmer equation and Lineweaver-Burk equation, it was found that BSA had reacted with naphthol green B and formed a new compound, the quenching action was due to static fluorescence quenching, and the action force was electrostatic interaction. According to the Lineweaver-Burk equation and thermodynamic equation, the average value of the binding constant (KLe: 1.411 x 10(5) L x mol(-1)), the thermodynamic parameters (DeltaHtheta: -5.707 kJ x mol(-1), DeltaGtheta: -30.25 kJ x mol(-1) and DeltaStheta: 79.95 J x K(-1)) and the amounts of binding sites (1.258) were obtained, providing important information for the research on the configuration modification of BSA because of the added naphthol green B, biological effects in a living body, and the coloration mechanism of naphthol green B.
Assuntos
Corantes/química , Compostos Férricos/química , Naftalenossulfonatos/química , Naftóis/química , Soroalbumina Bovina/química , Animais , Bovinos , Ligação Proteica , Espectrometria de FluorescênciaRESUMO
OBJECTIVE: To study the reconstructive effect of the dissociate bone flap to repair the macrosis depressed skull fracture on the frontal and orbit part. METHODS: The coronal scalp flap was elevated and dissociate bone flap was expanding to the 2cm width beside the edge of depressed skull fracture. The first step was to extract the dissociate bone flap and make there is an area for operating . Then extract free bone fragments, and elevate the depressed orbital lamina and use the biological glue to stick it to its position. The free fragments extracted were stacked into a whole one and it to its position in use of the biological glue on the dissociate bone flap. The uneven inner table should was smoother with bon-wax. The prosthetic dissociate bone flap was put back on its position and fixation. RESULTS: From January 2000 to December 2004, 17 cases of the macrosis depressed skull fracture on the frontal and orbit part undertaken plastic surgery by the dissociate bone flap to treat the macrosis depressed skull fracture and obtained excellent curative effect. CONCLUSIONS: Using dissociate bone flap to treat the mocrosis depressed skull fracture on the frontal and orbit part can avoid the complication of the traditional operation, and make the method become a plastic surgical operation.