RESUMO
Vascular calcification is extremely common and associated with major adverse cardiovascular events. Fibroblast growth factor (FGF) 21 has been identified as a potent metabolic regulator and a protector of the cardiovascular system. In this study, we aimed to investigate the effect of FGF21 on calcification of vascular smooth muscle cell (VSMC) and its mechanism. FGF21 inhibited beta-glycerophosphate (BGP) induced mineralization in VSMCs as determined by calcium concentration and Alizarin Red S. FGF21 suppressed BGP-induced BMP2/Smad signaling pathway components as well as osteoblast differentiation markers. FGF21 and Noggin could synergistically inhibit BGP-induced BMP2/Smad pathway expressions and calcification. Taken together, FGF21 inhibits vascular calcification in vitro by modulating BMP2/Smad signaling pathway.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Fatores de Crescimento de Fibroblastos/farmacologia , Miócitos de Músculo Liso/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Proteínas Smad/metabolismo , Calcificação Vascular/prevenção & controle , Animais , Western Blotting , Proteína Morfogenética Óssea 2/genética , Cálcio/metabolismo , Células Cultivadas , Expressão Gênica/efeitos dos fármacos , Músculo Liso Vascular/citologia , Miócitos de Músculo Liso/metabolismo , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/genética , Proteínas Smad/genética , Calcificação Vascular/genética , Calcificação Vascular/metabolismoRESUMO
Vascular calcification is prevalent and associated with adverse outcome without available therapy. The benefits of fibroblast growth factor (FGF)-21 on metabolism and atherosclerosis make it a promising therapeutic agent for vascular calcification. We investigated the effects of FGF21 on vascular smooth muscle cell (VSMC) calcification by culturing rat VSMCs in a calcifying medium for 9days. FGF21 markedly attenuated mineral deposition and apoptosis at the indicated time points. In the presence of FGF21, the expression levels of osteoblastic protein including bone morphogenic protein-2, alkaline phosphatase(ALP), runt-related transcription factor(RUNX)-2 and nuclear factor-kappa B ligand (RANKL) were down-regulated, whereas the expression of osteoprotegerin (OPG) increased. Knockdown of OPG significantly impaired inhibition of FGF21 on apoptosis and the expression of pro-apoptotic genes including caspase-3 and Bax and osteoblastic -promoting markers including ALP, RUNX-2 and RANKL. Furthermore, FGF21 facilitated the phosphoryl of AKT but suppressed P38, while OPG knockdown attenuated the effects. LY29400 (inhibitor of PI3K) abrogated the activation of PI3K/AKT and SB203580 (inhibitor of P38) abolished the inhibition of FGF21 on P38, while alteration was observed in the expression of RUNX-2. FGF21 inhibited VSMCs calcification via OPG/RANKL system, and through P38 andPI3K/AKT pathways.
Assuntos
Fatores de Crescimento de Fibroblastos/metabolismo , Osteoprotegerina/metabolismo , Ligante RANK/metabolismo , Calcificação Vascular/metabolismo , Calcificação Vascular/patologia , Animais , Apoptose , Células Cultivadas , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVES: There is increasing evidence that dual antiplatelet therapy (DAPT) when compared with single antiplatelet therapy may improve venous graft patency after coronary artery bypass graft. However, it is not yet known whether postoperative administration of DAPT may increase the potential risk of bleeding, especially in the early postoperative period. METHODS: We searched studies on PubMed, Embase, Web of Science and the Cochrane Central Register of Controlled Trials. Relative risk (RR) was pooled with 95% confidence intervals (CIs) for dichotomous data. Prior subgroup analyses were performed to look for potential heterogeneity. RESULTS: Thirteen studies involving 23 591 participants were included. Our meta-analysis showed that DAPT does not increase the risk of major bleeding (randomized controlled trials group: RR = 1.28, 95% CI 0.95-1.71; cohort studies group: RR = 0.99, 95% CI 0.66-1.51) and minor bleeding (randomized controlled trials group: RR = 1.15, 95% CI 0.73-1.81; cohort studies group: RR = 0.84, 95% CI 0.37-1.93) when compared with single antiplatelet therapy. Meanwhile, DAPT does not increase the incidence of major bleeding events during hospitalization (randomized controlled trials group: RR = 1.27, 95% CI 0.91-1.78; cohort studies group: RR = 0.50, 95% CI 0.12-2.09). Sensitivity analyses showed that our results are stable, and there was no evidence of publication bias. CONCLUSIONS: DAPT does not increase the risk of major bleeding and minor bleeding when compared with single antiplatelet therapy. Postoperative administration of DAPT is considered to be safe in patients after coronary artery bypass graft, even in the early postoperative period.
Assuntos
Ponte de Artéria Coronária , Inibidores da Agregação Plaquetária/uso terapêutico , Cuidados Pós-Operatórios/métodos , Hemorragia Pós-Operatória/epidemiologia , Saúde Global , Humanos , IncidênciaRESUMO
Nanoscale delivery systems have been widely investigated to overcome the penetration barrier of stratum corneum for effective transcutaneous application. The aim of this study is the development of effective vesicular formulations of ovalbumin and saponin which are able to promote penetration through the skin layers. Three kinds of vesicular formulations have been investigated as carriers, including liposomes, transfersomes and ethosomes, in which cholesterol and/or cationic lipid stearylamine are incorporated. The impact of membrane composition variations on the protein entrapment has been evaluated for each vesicle type. Formulations were characterized for particle size, polydispersity and encapsulation efficiency. The best formulations for each type of vesicle were subjected to in vivo transdermal immunization in mice. Among the three kinds of vesicular carrier, ethosomal nano carrier not only showed the best stability over a two months' storage, but also enabled the highest increase in the titer of serum antibody. In this regard, cationic nano-ethosomes can be considered as a promising vesicular carrier for transdermal vaccines. Meanwhile, we have developed a simple method to determine encapsulation efficiency of vesicular systems, which has potential application as a high throughput screening for vesicular formulations.
Assuntos
Sistemas de Liberação de Medicamentos , Nanopartículas , Ovalbumina/administração & dosagem , Saponinas/administração & dosagem , Administração Cutânea , Aminas/química , Animais , Química Farmacêutica/métodos , Colesterol/química , Portadores de Fármacos/química , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Feminino , Lipídeos/química , Lipossomos , Camundongos , Ovalbumina/farmacocinética , Ovalbumina/farmacologia , Tamanho da Partícula , Saponinas/farmacocinética , Saponinas/farmacologia , Pele/metabolismo , Absorção CutâneaRESUMO
OBJECTIVES: Vascular calcification is a dysfunction of the vasculature. Recent findings indicate that fibroblast growth factor21 (FGF21), a protector of the cardiovascular system, is related to the mineral deposition of bone and enhances the osteogenic activity of bone morphogenic protein (BMP)-2. In this study, we explored whether FGF21 suppresses vascular calcification. METHODS: A calcifying model was established by culturing primary rat vascular aortic smooth muscle cells (VSMCs) in a beta-glycerophosphate (BGP)-containing calcifying medium for 14 days. In addition, recombinant human FGF21 was applied to protect against VSMC calcification. RESULTS: In the presence of BGP, the expression levels of osteoblastic genes, including alkaline phosphatase (ALP), BMP-2 and runt-related transcription factor (RUNX)-2, were significantly upregulated on day 3, an effect that was maintained through day 14 (P < 0.001). A concomitant increase in ALP protein expression was observed through day 9 (P < 0.05). The incubation of VSMCs with calcifying medium for 14 days increased ALP activity (P < 0.05) and led to the formation of visible calcium nodules over the course of the protocol. ß-klotho expression was unaltered in BGP-induced VSMCs for the 14-day culture period. The culturing of VSMCs with calcifying medium led to opposing trends in the expression of FGFRs, namely, an increase in FGFR1 and FGFR4 mRNA levels (P < 0.001) and a decrease in FGFR2 and FGFR3 mRNA levels (P < 0.01). Reduced mineral deposition, in combination with decreased ALP activity (P < 0.001) and ALP protein expression (P < 0.001), was noted in VSMCs treated with varying doses of FGF21 and BGP in a dose-dependent manner. In addition, FGF21 downregulated osteoblastic-promoting gene expression, including ALP (P < 0.001), BMP-2 (P < 0.001) and RUNX-2 (P < 0.001). Furthermore, FGF21 enhanced ß-klotho expression (P < 0.05) and increased FGFR1 and FGFR3 mRNA levels (P < 0.001). FGFR-1 inhibitor SU5402 blocked partial inhibition of FGF21 on the expression of BMP-2 (P < 0.001) and RUNX-2 (P < 0.05). Furthermore, FGF21 suppressed the phosphorylation of P38, while P38 inhibitor, SB203580, attenuated the downregulation of RUNX-2 (P < 0.05). CONCLUSIONS: These data demonstrate FGF21 attenuates VSMC calcification in vitro via an FGF21/FGFR1/3/ß-klotho/P38MAPK/RUNX-2 signalling pathway.