TRPV5 attenuates abdominal aortic aneurysm in mice by regulating KLF4-dependent phenotype switch of aortic vascular smooth muscle cells.
TRPV5 attenuates abdominal aortic aneurysm in mice by regulating KLF4-dependent phenotype switch of aortic vascular smooth muscle cells.
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DOI：
10.1016/j.abb.2020.108724
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发表时间：
2020-12
影响因子：
3.9
通讯作者：
Shuo Wang;X. Tian;Dan Liu;Xiao-lin Zhang;Chenghui Yan;Yaling Han
中科院分区：
文献类型：
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作者：
Shuo Wang;X. Tian;Dan Liu;Xiao-lin Zhang;Chenghui Yan;Yaling Han
腹主动脉瘤（AAA）是一种致命的血管疾病，具有隐匿的症状。然而，其发展背后的机制仍不清楚。瞬时受体电位香草酸 (TRPV) 家族对心血管疾病具有重要的保护作用，但 TRPV5 在 AAA 中的作用尚未见报道。在这项研究中，ApoE−/−小鼠腹腔注射AAV-GFP或AAV-TRPV5。 30天后，小鼠进一步使用渗透泵给予血管紧张素II（Ang II，1.44 mg/kg/天）诱导AAA模型或盐水28天（即建立Saline + AAV-GFP、Saline + AAV-TRPV5、Ang II + AAV-GFP和Ang II + AAV-TRPV5组）。与对照组相比，28天时血管超声检测Ang II+AAV-TRPV5组AAA发生率和腹主动脉最大直径明显下降。同时，通过使用 Masson 和弹性蛋白染色，在 Ang II + AAV-TRPV5 组中观察到较少的胶原蛋白和弹性蛋白降解。此外，通过免疫组织化学在第 28 天收集的腹主动脉中观察到更多的 α-SMA 和更少的 MMP2。 在体外，用 Ang II (1 μM) 处理原代小鼠血管平滑肌细胞 (VSMC) 以诱导表型转换。 Sh-TRPV5和AdTRPV5用于转染VSMC。 PCR和Western blotting用于检测收缩标记物的表达，包括α-SMA和SM-22α。结果表明，α-SMA和SM-22α的mRNA和蛋白水平在Ang II的刺激下降低，但可以通过TRPV5过表达而减弱。细胞划痕实验表明，Ang II治疗组VSMCs的迁移能力增加，并且TRPV5过表达可以改善VSMCs的迁移能力。最重要的是，VSMC 在 Ang II 刺激下从收缩表型转变为分泌表型，但可以通过 TRPV5 过表达来挽救。此外，TRPV5 过表达抑制了 VSMC 中 Ang II 处理诱导的 KLF4 表达增加。数据表明TRPV5可以抑制AAA形成，并通过下调KLF4在VSMC表型转换中发挥关键作用，表明TRPV5可以作为治疗AAA的新策略。
Abdominal aortic aneurysm (AAA) is a fatal vascular disease with insidious symptoms. However, the mechanism behind its development remains unclear. The transient receptor potential vanilloid (TRPV) family has crucial protective effects against cardiovascular diseases, but the role of TRPV5 in AAA has yet to be reported. In this study, ApoE−/−mice were intraperitoneally injected with AAV-GFP or AAV-TRPV5. After 30 days, mice were further administered with angiotensin II (Ang II, 1.44 mg/kg/day) by using osmotic pumps to induce the AAA model or Saline for 28 days, (i.e., Saline + AAV-GFP, Saline + AAV-TRPV5, Ang II + AAV-GFP and Ang II + AAV-TRPV5 groups were established). Compared with the control group, the incidence of AAA and the maximal diameter of the abdominal aorta markedly decreased in Ang II + AAV-TRPV5, which was detected by vascular ultrasound at 28 day. Meanwhile, less collagen and elastin degradation were observed in the Ang II + AAV-TRPV5 group by using Masson and Elastin stains. Moreover, more α-SMA and less MMP2 was observed in the abdominal aortas collected at 28 day by immunohistochemistry.In vitro, primary mouse vascular smooth muscle cells (VSMCs) were treated with Ang II (1 μM) to induce phenotype switch. Sh-TRPV5 and AdTRPV5 were used to transfect VSMCs. PCR and Western blotting were used to access the expression of contractile marker, including α-SMA and SM-22α. The results showed that the mRNA and protein level of α-SMA and SM-22α were decreased under the stimulation of Ang II, but could be attenuated by TRPV5 overexpression. The cell scratch assay demonstrated that the migration ability of VSMCs was increased in Ang II treated group and could be ameliorated by TRPV5 overexpression. Above all, VSMCs transformed from the contractile into secretory phenotype under Ang II stimuli, but could be rescued by TRPV5 overexpression. Furthermore, TRPV5 overexpression suppressed the increased expression of KLF4 induced by Ang II treatment in VSMCs. The data demonstrated that TRPV5 could inhibit AAA formation and play a critical role in the VSMC phenotype switch by downregulating KLF4, suggesting TRPV5 as a new strategy for treating AAA.