成纤维细胞生长因子21在体外可减轻血管平滑肌细胞的钙化。
Fibroblast growth factor 21 attenuates calcification of vascular smooth muscle cells in vitro.
作者信息
Cao Fangying, Wang Shaoping, Cao Xiangrong, Liu Xiaoxiao, Fu Kun, Hao Peng, Liu Jinghua
机构信息
Department of Cardiology, Beijing Anzhen Hospital, Beijing Institute of Heart, Lung and Blood Vessel Diseases, Capital Medical University, Beijing, China.
Department of Cardiac Surgery, Beijing Anzhen Hospital, Beijing Institute of Heart, Lung and Blood Vessel Diseases, Capital Medical University, Beijing, China.
出版信息
J Pharm Pharmacol. 2017 Dec;69(12):1802-1816. doi: 10.1111/jphp.12826. Epub 2017 Oct 4.
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. β-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 β-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/β-klotho/P38MAPK/RUNX-2 signalling pathway.
目的
血管钙化是血管系统的一种功能障碍。最近的研究结果表明，成纤维细胞生长因子21（FGF21）作为心血管系统的一种保护因子，与骨骼的矿物质沉积有关，并增强骨形态发生蛋白（BMP）-2的成骨活性。在本研究中，我们探讨了FGF21是否能抑制血管钙化。
方法
通过在含β-甘油磷酸（BGP）的钙化培养基中培养原代大鼠血管主动脉平滑肌细胞（VSMC）14天，建立钙化模型。此外，应用重组人FGF21来预防VSMC钙化。
结果
在存在BGP的情况下，成骨基因的表达水平，包括碱性磷酸酶（ALP）、BMP-2和 runt相关转录因子（RUNX）-2，在第3天显著上调，这种作用一直持续到第14天（P < 0.001）。在第9天观察到ALP蛋白表达伴随性增加（P < 0.05）。用钙化培养基培养VSMC 14天增加了ALP活性（P < 0.05），并在实验过程中导致可见钙结节的形成。在为期14天的培养期内，BGP诱导的VSMC中β-klotho表达未改变。用钙化培养基培养VSMC导致FGFRs表达出现相反的趋势，即FGFR1和FGFR4 mRNA水平增加（P < 0.001），而FGFR2和FGFR3 mRNA水平降低（P < 0.01）。在用不同剂量的FGF21和BGP处理的VSMC中，观察到矿物质沉积减少，同时ALP活性（P < 0.001）和ALP蛋白表达（P < 0.001）呈剂量依赖性降低。此外，FGF21下调促进成骨的基因表达，包括ALP（P < 0.001）、BMP-2（P < 0.001）和RUNX-2（P < 0.001）。此外，FGF21增强β-klotho表达（P < 0.05），并增加FGFR1和FGFR3 mRNA水平（P < 0.001）。FGFR-1抑制剂SU5402阻断了FGF21对BMP-2（P < 0.001）和RUNX-2（P < 0.05）表达的部分抑制作用。此外，FGF21抑制P38的磷酸化，而P38抑制剂SB203580减弱了RUNX-2的下调（P < 0.05）。
结论
这些数据表明，FGF21通过FGF21/FGFR1/3/β-klotho/P38MAPK/RUNX-2信号通路在体外减弱VSMC钙化。
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