RESUMEN
This study explores the effect of apigenin(APG), oxymatrine(OMT), and APG+OMT on the proliferation of non-small cell lung cancer cell lines and the underlying mechanisms. Cell counting kit-8(CCK-8) assay was used to detect the vitality of A549 and NCI-H1975 cells, and colony formation assay to evaluate the colony formation ability of the cells. EdU assay was employed to examine the proliferation of NCI-H1975 cells. RT-qPCR and Western blot were performed to detect the mRNA and protein expression of PLOD2. Molecular docking was carried out to explore the direct action ability and action sites between APG/OMT and PLOD2/EGFR. Western blot was used to study the expression of related proteins in EGFR pathway. The viability of A549 and NCI-H1975 cells was inhibited by APG and APG+OMT at 20, 40, and 80 μmol·L~(-1) in a dose-dependent manner. The colony formation ability of NCI-H1975 cells was significantly suppressed by APG and APG+OMT. The mRNA and protein expression of PLOD2 was significantly inhibited by APG and APG+OMT. In addition, APG and OMT had strong binding activity with PLOD2 and EGFR. In APG and APG+OMT groups, the expression of EGFR and proteins in its downstream signaling pathways was significantly down-regulated. It is concluded that APG in combination with OMT could inhibit non-small lung cancer, and the mechanism may be related to EGFR and its downstream signaling pathways. This study lays a new theoretical basis for the clinical treatment of non-small cell lung cancer with APG in combination with OMT and provides a reference for further research on the anti-tumor mechanism of APG in combination with OMT.
Asunto(s)
Humanos , Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Apigenina , Simulación del Acoplamiento Molecular , Alcaloides , Quinolizinas , ARN Mensajero , Receptores ErbBRESUMEN
Background Cluster of differentiation 40 ligand (CD40L) and rosuvastatin (RSV) affect atherosclerotic plaque stability, but little is known about their roles in extracellular matrix (ECM) production. We investigated the effects of CD40L and RSV on pre-existing advanced plaques. Methods and results Pre-existing advanced plaques were induced in apolipoprotein E-knockout (ApoE-/-) mice by the surgical placement of carotid constrictive silastic collars. Two weeks after surgery, mice were divided into the following treatment groups: control, empty adenovirus, CD40L adenovirus, CD40L adenovirus + RSV, and RSV. Mice received adenovirus via two tail-vein injections (2 × 109 pfu each) and/or RSV via intragastric administration (5 mg/kg; daily for 4 weeks). Mice in the CD40L adenovirus group exhibited increased plaque disruption rates, increased relative plaque macrophage and lipid content, reduced plaque collagen content, and increased local inflammation compared to the other treatment groups, but no significant differences in plaque area were observed among the groups. Notably, in the atherosclerotic plaques of the CD40L adenovirus group, both the mRNA and protein expression of prolyl-4-hydroxylase alpha 1 (P4Hα1) was significantly decreased, leading to a consequent decrease in the protein expression of collagen types I and III. Treatment with RSV decreased the serum levels of CD40L in a lipid-independent fashion and attenuated the effects of CD40L overexpression, particularly with respect to P4Hα1 downregulation. Conclusions CD40L destabilized advanced plaques in the carotid arteries of ApoE-/- mice, in part by decreasing P4Hα1 expression, and consequently collagen expression. These destabilizing effects were attenuated by RSV.
Asunto(s)
Apolipoproteínas E/deficiencia , Ligando de CD40/genética , Placa Aterosclerótica/tratamiento farmacológico , Procolágeno-Prolina Dioxigenasa/genética , Rosuvastatina Cálcica/farmacología , Animales , Apolipoproteínas E/genética , Ligando de CD40/sangre , Modelos Animales de Enfermedad , Regulación hacia Abajo/efectos de los fármacos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Lípidos/sangre , Masculino , Ratones , Ratones Noqueados para ApoE , Placa Aterosclerótica/genética , Placa Aterosclerótica/patología , Procolágeno-Prolina Dioxigenasa/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
The aim of this paper was to find out the active components of Epimedium brevicornum using network pharmacology, and find the potential targets and mechanisms. The TCMSP database was used to screen the active ingredients, and TTD and DrugBank databases were used to predict the potential targets with the literature mining. The pathway annotation was used to enrich and analyze the active ingredients and potential targets of E. brevicornum. The results showed that E. brevicornum had34 potential target active ingredients, including 21 flavones components, such as icariin, epimedin A, epimedin B, epimedin C, Yinyanghuo A, Yinyanghuo C and so on, 2 lignans involved in (+)-cycloolivil and olivil, 3 sterols consisting of sitosterol, 24-epicampesterol and poriferast-5-en-3beta-ol. The main predicted targets included Ptgs2, NCOA6, RANK, OPG, WNT9B, PTH1R, BMPs, SMAD4A and so on. There were 88 signaling pathways involved in 10 signaling pathways which was related to inflammation, such as NF-kappa B signaling pathway, T cell receptor signaling pathway, IL-17 signaling pathway and 10 pathways which was related to cancer included breast cancer, bladder cancer, pancreatic cancer and so on, and estrogen related signaling pathways included estrogen signaling pathway. This laid the foundation for the discovery of the active components of Epimedium and the study on its mechanism of action.
Asunto(s)
Epimedium/metabolismo , Epimedium/clasificación , Estrógenos , Flavonoides , Transducción de SeñalRESUMEN
Endoplasmic reticulum stress (ERs) has been regarded as an important cause for the pathogenesis of non-small-cell lung cancer (NSCLC). ß-elemene is an active component in the essential oil extracted from a medicinal herb, Curcuma wenyujin, and has been reported to be effective against non-small-cell lung cancer (NSCLC). However, the potential effect and underlying mechanisms of ß-elemene on regulating ERs to inhibit NSCLC are still unclear. In the present study, A549 cells and Lewis tumor-bearing C57BL/6J mice were established to evaluate this effect. Visualsonics Vevo 2100 Small Animal Dedicated High-frequency Color Ultrasound was performed to observe tumor volume in vivo. 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) was used to evaluate cell vitality of A549 cells. Furthermore, western blotting (WB), immunohistochemistry (IHC) and quantitative reverse transcription polymerase chain reaction (q-PCR) were applied to detect the ERs-related proteins. Flow cytometry was also applied to detect cell apoptosis and assay kit for reactive oxygen species (ROS) generation. Our results showed that ß-elemene inhibited lung cancer tumor growth and cell vitality in a dose- and time-dependent manner. Not only that, ß-elemene could up-regulate ERs-related proteins like PERK, IRE1α, ATF6, ATF4, CHOP and down-regulate the Bcl-2 expression. More importantly, ERs inhibitor 4-PBA, IRE1α inhibitor STF-083010, ATF6 inhibitor Anti-ATF6 and PERK inhibitor GSK2656157 can all reduce the amplitude of protein expression changes and apoptosis rates, then weaken the anti-tumor effect of ß-elemene. Therefore, the present in vivo and in vitro study revealed that the anti-NSCLC effect of ß-elemene is closely related to the activation of ERs through PERK/IRE1α/ATF6 pathway, and this might be beneficial for clinical therapy of NSCLC.
Asunto(s)
Factor de Transcripción Activador 6/metabolismo , Apoptosis/efectos de los fármacos , Carcinoma de Pulmón de Células no Pequeñas/patología , Estrés del Retículo Endoplásmico/efectos de los fármacos , Endorribonucleasas/metabolismo , Neoplasias Pulmonares/patología , Proteínas Serina-Treonina Quinasas/metabolismo , Sesquiterpenos/farmacología , eIF-2 Quinasa/metabolismo , Células A549 , Animales , Antineoplásicos Fitogénicos/farmacología , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Supervivencia Celular/efectos de los fármacos , Humanos , Neoplasias Pulmonares/metabolismo , Ratones Endogámicos C57BL , Modelos Biológicos , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Factores de Tiempo , Factor de Transcripción CHOP/metabolismoRESUMEN
The present study reports a case of electrical storm occurring in a 43-year-old woman with dilated cardiomyopathy. The patient suffered from a cardiac electrical storm, with 98 episodes of ventricular tachycardia rapidly degenerating to ventricular fibrillation in hospital. The patient was converted with a total of 120 defibrillations. Recurrent ventricular tachycardia/fibrillation was initiated by premature ventricular beats. The patient did not respond to the use of amiodaronum. However, the administration of esmolol stabilized the patient's heart rhythm. A moderate dose of the ß-blocker esmolol, administered as an 0.5-mg intravenous bolus injection followed by an infusion at a rate of 0.15 mg/kg/min, inhibited the recurrence of ventricular fibrillation and normalized the electrocardiographic pattern. The results suggest that esmolol may be able to improve the survival rate of patients with electrical storm in dilated cardiomyopathy and should be considered as a primary therapy in the management of cardiac electrical storms.
RESUMEN
CD40L and statins exhibit pro-inflammatory and anti-inflammatory effects, respectively. They are both pleiotropic and can regulate extracellular matrix (ECM) degeneration in an atherosclerotic plaque. Statins can decrease both the CD40 expression and the resulting inflammation. However, the effects of CD40L and stains on atherosclerotic plaque ECM production and the underlying mechanisms are not well established. Moreover, prolyl-4-hydroxylase α1 (P4Hα1) is involved in collagen synthesis but its correlations with CD40L and statins are unknown. In the present study, CD40L suppressed P4Hα1 expression in human aortic smooth muscle cells (HASMCs) in a dose- and time-dependent manner, with insignificant changes in MMP2 expression and negative enzymatic activity of MMP9. CD40L increased TRAF6 expression, JNK phosphorylation, NF-κB nuclear translocation as well as DNA binding. Furthermore, silencing TRAF6, JNK or NF-κB genes abolished CD40L-induced suppression of P4Hα1. Lower NF-κB nuclear import rates were observed when JNK or TRAF6 silenced HASMCs were stimulated with CD40L compared to HASMCs with active JNK or TRAF6. Together, these results indicate that CD40L suppresses P4Hα1 expression in HASMCs by activating the TRAF6-JNK- NF-κB pathway. We also found that rosuvastatin inhibits CD40L-induced activation of the TRAF6-JNK- NF-κB pathway, thereby significantly rescuing the CD40L stimulated P4Hα1 inhibition. The results from this study will help find potential targets for stabilizing vulnerable atherosclerotic plaques.
Asunto(s)
Aorta/efectos de los fármacos , Ligando de CD40/efectos de los fármacos , Regulación hacia Abajo/fisiología , Matriz Extracelular/metabolismo , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Músculo Liso Vascular/efectos de los fármacos , Rosuvastatina Cálcica/farmacología , Aorta/citología , Aorta/metabolismo , Ligando de CD40/fisiología , Células Cultivadas , Humanos , MAP Quinasa Quinasa 4/metabolismo , Músculo Liso Vascular/citología , Músculo Liso Vascular/metabolismo , FN-kappa B/metabolismo , Fosforilación , Factor 6 Asociado a Receptor de TNF/metabolismoRESUMEN
MicroRNAs have become recognized as key players in the development of malignancy. They are a family of small non-coding RNAs (22 nt~30 nt) that can negatively regulate the expression of cancer-related genes by sequence selective targeting of mRNAs, leading to either mRNA translational repression or degradation. Lung cancer is the leading cause of cancer-related death worldwide with a substantially low survival rate. In this study, we analyzed the expression profile of miR-34c-3p in non-small cell lung cancer (NSCLC) tissues and cell lines, as its participation in some other types of cancer has been shown by previous reports. We found that miR-34c-3p was downregulated both in NSCLC tissues and cell lines. Overexpression of miR-34c-3p suppressed cell proliferation and colony formation and also limited migration and invasion in A549 cells. Furthermore, our results also shown miR-34c-3p reduction was associated with increased PAC1 expression levels in which miR-34c-3p downregulated PAC1 expression by recognizing and binding to specific binding sites in PAC1 3'-UTR. Taken together, our study implicates important roles of miR-34c-3p in NSCLC pathogenesis and implicates its potential application in cancer therapy.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/patología , Proliferación Celular , Regulación Neoplásica de la Expresión Génica/genética , Neoplasias Pulmonares/patología , MicroARNs/genética , Transducción de Señal , Anciano , Western Blotting , Carcinoma de Pulmón de Células no Pequeñas/genética , Línea Celular Tumoral , Movimiento Celular/genética , Fosfatasa 2 de Especificidad Dual/metabolismo , Femenino , Humanos , Inmunohistoquímica , Neoplasias Pulmonares/genética , Sistema de Señalización de MAP Quinasas/fisiología , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Reacción en Cadena en Tiempo Real de la Polimerasa , Transcriptoma , TransfecciónRESUMEN
In the current study, finite element analyses were conducted to examine the biomechanical capability of a newly design dynamic stabilization system, FlexPLUS, to restore the load transmission of degenerated intervertebral L4-L5 lumbar motion segment spine under compression. Detailed three-dimensional FE models of L4-L5 motion segment and the FlexPLUS were developed. Compressive loading up to 1000N was applied to the intact L4-L5 model, the L4-L5 models with slight and moderate degenerated disc, and the implanted L4-L5 model. Further more, the load transmission characteristics of Dynesys and a rigid rod was also simulated for comparison. The resultant load-displacement curves and the load transferred through annulus under various conditions were compared. The predicted axial displacement of L4 top surface against applied compressive force of the intact L4-L5 model agreed well with experimental data. The predicted results showed that degenerated disc has significant effect on the lumbar segment load bearing capacity. Not only the stiffness of the segment was greatly increased, the uniform nature of the disc stress distribution was also altered. The FlexPLUS can effectively reduce the disc loading of degenerated model. Although the non-uniform load distribution pattern through annulus was not improved, the overall stress magnitude was greatly reduced to the level of intact model for grade II degeneration.