Sonic hedgehog promotes autophagy of vascular smooth muscle cells
Sonic hedgehog promotes autophagy of vascular smooth muscle cells
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DOI：
10.1152/ajpheart.00160.2012
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发表时间：
2012-12-01
影响因子：
4.8
通讯作者：
Zheng, Xi-Long
中科院分区：
文献类型：
--
作者：
Li, Haijie;Li, Jingjing;Zheng, Xi-Long
李辉，李军，李毅，辛格平，曹磊，徐磊，李东，王勇，谢志，桂勇，郑旭。Sonic hedgehog基因促进血管平滑肌细胞自噬。[J] .中国生物医学工程学报，2016,31(2):557 - 557。首次发表于2012年9月28日；doi: 10.1152 / ajpheart.00160.2012。-Sonic hedgehog （Shh）是一种形态因子，在动脉粥样硬化病变中被重新表达。它还能刺激血管平滑肌细胞（SMCs）的增殖。血管SMC的自噬可以促进SMC的存活并增加斑块的稳定性。本研究的目的是探讨Shh是否诱导血管SMCs自噬。我们的研究表明，Shh蛋白和微管相关蛋白1轻链3 (LC3)-II在小鼠颈总动脉内膜病变的SMCs中升高。在培养的小鼠主动脉SMCs中，重组小鼠Shh刺激LC3-II水平。通过四环素调控的表达诱导系统，野生型小鼠Shh在人主动脉SMCs中过表达，时间依赖性地增加LC3-II水平，并刺激蛋白激酶B （AKT）磷酸化。AKT抑制剂IV （AKTI IV）预处理可抑制AKT磷酸化和LC3-II的升高。免疫染色和透射电镜检测发现，Shh诱导的自噬体的形成被AKTI IV抑制。在巴霉素A1、(2S, 3S)-反式环氧琥珀酰-l -乙酰氨基-3-甲基丁烷乙酯、胃抑素A或自噬相关基因7 (ATG7) siRNA的存在下，Shh进一步增加SMC LC3-II。此外，Shh诱导SMC增殖，不仅可以被AKTI IV抑制，还可以被Shh受体抑制剂环巴胺抑制。3-甲基腺嘌呤（3-MA）、巴菲霉素A1或ATG7 siRNA抑制自噬导致细胞增殖受到抑制。用3-MA、AKTI IV或环巴胺治疗可抑制小鼠颈总动脉新生内膜的形成。综上所述，我们的研究结果表明Shh诱导血管SMCs自噬涉及AKT激活，提示自噬在Shh诱导的细胞反应中起作用。
Li H, Li J, Li Y, Singh P, Cao L, Xu L, Li D, Wang Y, Xie Z, Gui Y, Zheng X. Sonic hedgehog promotes autophagy of vascular smooth muscle cells. Am J Physiol Heart Circ Physiol 303: H1319-H1331, 2012. First published September 28, 2012; doi: 10.1152/ajpheart.00160.2012.-Sonic hedgehog (Shh) is a morphogen critically involved in development that is reexpressed in atherosclerotic lesions. It also stimulates proliferation of vascular smooth muscle cells (SMCs). Autophagy in vascular SMCs is known to promote SMC survival and increase plaque stability. The aim of this study was to investigate whether Shh induces autophagy of vascular SMCs. Our study showed that both Shh protein and microtubule-associated protein 1 light chain 3 (LC3)-II were increased in SMCs within neointimal lesions of mouse common carotid arteries. In cultured mouse aortic SMCs, recombinant mouse Shh stimulated LC3-II levels. Overexpression of wild-type mouse Shh through the tetracycline-regulated expression-inducible system in human aortic SMCs time-dependently increased the levels of LC3-II and also stimulated protein kinase B (AKT) phosphorylation. Pretreatment with AKT inhibitor IV (AKTI IV) inhibited AKT phosphorylation and the increase in LC3-II. Shh-induced autophagy was further confirmed by the formation of autophagosomes as detected by immunostaining and transmission electron microscopy, which was inhibited by AKTI IV. Shh further increased SMC LC3-II in the presence of bafilomycin A1, (2S, 3S)-trans-epoxysuccinyl-L-leucylamido-3-methylbutane ethyl ester, and pepstatin A or siRNA for the autophagy-related gene 7 (ATG7). In addition, Shh induced SMC proliferation, which was inhibited not only by AKTI IV but also by cyclopamine, an inhibitor of Shh receptor. Inhibition of autophagy with 3-methyladenine (3-MA), bafilomycin A1, or ATG7 siRNA resulted in inhibition of cell proliferation. Treatment with 3-MA, AKTI IV, or cyclopamine inhibited neointima formation in mouse common carotid arteries. Taken together, our results have shown that Shh induces autophagy of vascular SMCs involving AKT activation, suggesting a role of autophagy in Shh-induced cellular responses.