RESUMO
Context: Scutellaria baicalensis Georgi (SBG) may relieve bone cancer pain (BCP) by regulating cell proliferation, angiogenesis, and apoptosis. Objective: The mechanism of SBG in the treatment of BCP remains to be further explored. Methods: The active compounds and targets of SBG were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and SwissTargetPrediction databases. BCP-related targets were screened from NCBI and GeneCards databases. Additionally, Cytoscape software was applied to construct network diagrams, and OmicShare platform was used to enrich Gene Ontology (GO) and pathways. Finally, the verification of active compounds and core targets was performed based on quantitative real-time PCR (qRT-PCR). Results: Interestingly, we identified baicalein and wogonin as the main active components of SBG. A total of 41 SBG targets, including VEGFA, IL6, MAPK3, JUN and TNF, were obtained in the treatment of BCP. In addition, pathways in cancer may be an essential way of SBG in the treatment of BCP. Experimental verification had shown that baicalein and wogonin were significantly related to BCP core targets. Conclusions: The active components of SBG have been clarified, and the mechanism of the active components in treating BCP has been predicted and verified, which provides an experimental and theoretical basis for the in-depth elucidation of the pharmacodynamics material basis and mechanism of SBG.
Assuntos
Dor do Câncer , Neoplasias , Scutellaria baicalensis , Farmacologia em Rede , Medicina Tradicional ChinesaRESUMO
Bone cancer pain (BCP) is the most common type of pain in cancer patients, during which microglia cells were activated. A previous study showed BAM8-22 had the ability to alleviate BCP via inhibiting microglia activation while the mechanism was not clear. This study aims to investigate the specific mechanism of BAM8-22 inhibiting microglia activation. This study was mainly investigated in BCP mice or LPS-treated microglia BV-2 cells. The behavior tests of mice were performed at 0, 1, 2, 12, and 24 h after BAM8-22 treatment. The expression of miR-184 and CX3CR1 mRNAs was detected by quantitative RT-PCR. The expression of CX3CR1 protein and microglia activation marker, Iba-1, was measured by western blot analysis. The levels of TNF-α and IL-1ß were detected by ELISA. Dual-luciferase assay was performed to verify the combination between miR-184 and CX3CR1. After BAM8-22 treatment, increased miR-184 level was observed in both BCP mice and LPS-treated BV-2 cells, with the downregulated expression of Iba-1 and inflammatory cytokines, namely the inhibition of microglia activation. The inhibition of miR-184 reversed the inhibitory effect of BAM8-22 on microglia activation. Further, in vitro studies showed that miR-184 bound to the 3'UTR of CX3CR1 and inhibited microglia activation via repressing CX3CR1 expression. What's more, the suppression of CX3CR1 expression eliminated the reversal effect of the miR-184 inhibitor on BAM8-22-induced microglia activation and decreased Iba-1 expression and pro-inflammatory cytokine secretion. In BCP models, miR-184 was upregulated by BAM8-22 and the elevated level of miR-184 bound to the 3'UTR region of CX3CR1 and repressed CX3CR1 expression, thus inhibiting the microglia activation, suggesting the potential application of miR-184/CX3CR1 for BCP treatment.
Assuntos
Anti-Inflamatórios/farmacologia , Receptor 1 de Quimiocina CX3C/genética , MicroRNAs/metabolismo , Microglia/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Animais , Receptor 1 de Quimiocina CX3C/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Células Cultivadas , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolissacarídeos/toxicidade , Masculino , Camundongos , MicroRNAs/genética , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Microglia/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Propofol has been found to play an important role in hepatic ischemia/reperfusion (I/R) injury with the antioxidant effects. However, the molecular mechanism of propofol in hepatic I/R injury has not been fully understood. Male Sprague-Dawley rats were randomly assigned into Sham group, hepatic I/R group, and propofol treatment group. I/R injury was attained by ischemia for 1 h and reperfusion for 2 h. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity were detected. QSG-7701 cells were cultured in hypoxia condition for 15 h and then in reoxygenation condition for 6 h to imitate hypoxia/reoxygenation (H/R) injury in vitro. Real-time RT-PCR and Western blot were performed to determine the expression of miR-133a-5p and MAPK6. Luciferase reporter assay was used to determine the regulation of miR-133a-5p on MAPK6. Propofol significantly reduced the activities of serum AST and ALT induced by hepatic I/R injury in rats. Propofol increased the level of miR-133a-5p and decreased the expression of MAPK6 in vivo and in vitro. Luciferase reporter assay showed that MAPK6 was a target of miR-133a-5p. Knockdown of miR-133a-5p abrogated the effect of propofol on the upregulation of MAPK6 induced by H/R. MAPK6 overexpression promoted the cell apoptosis induced by H/R which could be attenuated by propofol. Finally, we found that miR-133a-5p reversed the protective effect of propofol in rats with hepatic I/R injury. Propofol showed protective roles for hepatic I/R injury in vivo and H/R injury in vitro, which involved with miR-133a-5p regulating the expression of MAPK6.
