Levistilide A inhibits angiogenesis in liver fibrosis via vascular endothelial growth factor signaling pathway
Levistilide A inhibits angiogenesis in liver fibrosis via vascular endothelial growth factor signaling pathway
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Levistilide A通过血管内皮生长因子信号通路抑制肝纤维化中的血管生成
DOI：
10.1177/1535370217701005
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发表时间：
2017-03
影响因子：
3.2
通讯作者：
Cheng-Hai Liu
中科院分区：
文献类型：
--
作者：
Zhi-Min Zhao;Hong-Liang Liu;Xin Sun;Tao Guo;Li Shen;Yan-Yan Tao;Cheng-Hai Liu
当归（Angelica sinensis，Danggui）的Levistilide A（C24 H28 O 4，分子量= 380.48）具有抑制肝星状细胞增殖的作用。本研究探讨了来维昔单抗A对与血管生成相关的肝纤维化的影响，特别是对肝窦内皮细胞特征性变化的影响。用TGF-β1激活LX-2细胞，用内皮细胞生长素诱导人肝窦内皮细胞（humanhepaticsinusoidendothelialcells，HHSECs）。使用甲基噻唑二苯基四唑溴化物测定法检测细胞活力;通过荧光探针法观察F-肌动蛋白;使用EdU试剂盒检查细胞增殖;使用管形成测定法和转基因斑马鱼模型评估抗血管生成活性。为了验证体内结果，大鼠每周两次皮下注射CCl 4，持续6周以复制肝纤维化模型，然后从第4周起给予10 mL/kg生理盐水、4 mg/kg索拉非尼、3和6 mg/kg来维司他胺A治疗3周。天狼星红染色检测胶原沉积; vWF标记和X线二维成像检测肝脏微血管;扫描电镜观察肝窦开窗; Western blotting检测I型胶原、α-SMA、CD 31、血管内皮生长因子（VEGF）和VEGF-R2。我们的研究结果表明，levistilide A衰减LX-2细胞活化和HHSEC增殖。HHSECs在Matrigel中形成管状结构的能力受到抑制，转基因斑马鱼中功能性血管的数量减少。在体内实验中，levistilide A减少胶原沉积和新的微血管的数量，改善窦毛细血管化，并下调CD 31，VEGF和VEGF-R2的表达。这些发现表明，Levistilide A可以通过VEGF信号通路减轻肝窦毛细血管化，从而抑制肝纤维化，这是通过抗血管生成来实现的。在本研究中，我们进一步研究了levistilide A抗肝纤维化与血管生成有关的机制，特别是对肝窦内皮细胞特征性变化的影响。采用肝星状细胞、肝窦内皮细胞模型和四氯化碳诱导的肝纤维化模型。这些结果表明，levistilide A可以抑制肝纤维化，通过血管内皮生长因子信号通路，通过减轻血窦毛细血管化的抗血管生成。证实了来维司他胺A的抗肝纤维化作用，并对其具体机制进行了探讨。这些结果表明，levistilide A可能是一个很有潜力的通过抗血管生成治疗肝纤维化的药物，这种作用将在其他纤维化动物模型研究或临床试验中得到验证。
Levistilide A (C24H28O4, molecular weight = 380.48) derived fromAngelica sinensis(Danggui) has been reported to inhibit hepatic stellate cell proliferation. This study investigated the effects of levistilide A on liver fibrosis relating to angiogenesis, particularly on the characteristic change in liver sinusoidal endothelial cells. LX-2 cells were activated by TGF-β1, and the human hepatic sinusoidal endothelial cells (HHSECs) were induced by endothelial cell growth supplement. Cell viability was detected using a methylthiazoldiphenyl–tetrazolium bromide assay; F-actin was visualized through the fluorescence probe method; cell proliferation was examined using the EdU kit; antiangiogenesis activity was assessed using the tube formation assay and transgenic zebrafish model. To verify the resultsin vivo, rats were subcutaneously injected with CCl4twice a week for six weeks to duplicate the liver fibrosis model and then treated with 10 mL/kg of normal saline, 4 mg/kg of sorafenib, and 3 and 6 mg/kg of levistilide A for three weeks from the fourth week. Collagen deposition was detected through Sirius Red staining; liver microvasculature was examined through vWF labeling and X-ray 2D imaging; sinusoidal fenestrations were observed through scanning electron microscopy; collagen I, α-SMA, CD31, vascular endothelial growth factor (VEGF), and VEGF-R2 were detected through Western blotting. Our results indicated that levistilide A attenuated LX-2 cell activation and HHSEC proliferation. The ability of HHSECs to form tubelike structures in Matrigel was inhibited, and the number of functional vessels in transgenic zebrafish decreased. Inin vivoexperiments, levistilide A reduced collagen deposition and the number of new microvessels; ameliorated sinusoid capillarization; and downregulated the expression of CD31, VEGF, and VEGF-R2. These findings suggest that levistilide A can inhibit liver fibrosis through antiangiogenesis by alleviating sinusoid capillarization via the VEGF signaling pathway.Impact statementLevistilide A has been reported to inhibit hepatic stellate cell (HSC) proliferation. In this study, we further investigated the mechanisms of levistilide A on liver fibrosis relating to angiogenesis, particularly on the characteristic change in liver sinusoidal endothelial cells. The cell models of HSC and liver sinusoidal endothelial cell and CCl4induced liver fibrosis model were used. These results suggest that levistilide A can inhibit liver fibrosis through antiangiogenesis by alleviating sinusoid capillarization via the vascular endothelial growth factor signaling pathway. The effect of levistilide A on liver fibrosis was confirmed, and its detailed mechanism was also discussed. These findings suggest that levistilide A may be a great potential drug for treating liver fibrosis through antiangiogenesis, and this effect will be verified in other fibrotic animal model studies or by clinical trials.
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影响因子：
3.1
作者：
Hua Hong;Eun-Hee Kim;Ho J. Lee;Yoon Jae Kim;Jong Joon Lee;K. Hahm
通讯作者：
Hua Hong;Eun-Hee Kim;Ho J. Lee;Yoon Jae Kim;Jong Joon Lee;K. Hahm
影响因子：
13.5
作者：
DeLeve, Laurie D.
通讯作者：
DeLeve, Laurie D.
影响因子：
19.7
作者：
Hidezo Mori;K. Hyodo;E. Tanaka;M. Uddin-Mohammed;A. Yamakawa;Y. Shinozaki;Hiroe Nakazawa;Y. Tanaka;T. Sekka;Y. Iwata;S. Handa;K. Umetani;H. Ueki;T. Yokoyama;K. Tanioka;M. Kubota;H. Hosaka;N. Ishikawa;M. Ando
通讯作者：
Hidezo Mori;K. Hyodo;E. Tanaka;M. Uddin-Mohammed;A. Yamakawa;Y. Shinozaki;Hiroe Nakazawa;Y. Tanaka;T. Sekka;Y. Iwata;S. Handa;K. Umetani;H. Ueki;T. Yokoyama;K. Tanioka;M. Kubota;H. Hosaka;N. Ishikawa;M. Ando
DOI：
10.1159/000352074
发表时间：
2014
期刊：
Chemical immunology and allergy
影响因子：
--
作者：
G. Marone;F. Granata
通讯作者：
G. Marone;F. Granata
影响因子：
13.5
作者：
Straub, Adam C.;Stolz, Donna B.;Barchowsky, Aaron
通讯作者：
Barchowsky, Aaron