Knockdown of estrogen-related receptor α inhibits valve interstitial cell calcification in vitro by regulating heme oxygenase 1
Knockdown of estrogen-related receptor α inhibits valve interstitial cell calcification in vitro by regulating heme oxygenase 1
复制标题
雌激素相关受体α的敲低通过调节血红素氧合酶1抑制体外瓣膜间质细胞钙化
DOI：
10.1096/fj.202001588rr
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发表时间：
2020-11-13
期刊：
影响因子：
4.8
通讯作者：
Wang, Jian'an
中科院分区：
文献类型：
--
作者：
Hu, Wangxing;Wu, Rongrong;Wang, Jian'an
钙化性主动脉瓣疾病（CAVD）是成人最常见的心脏瓣膜病。CAVD的细胞机制尚不清楚，但越来越多的证据表明，人瓣膜间质细胞（hVIC）的成骨分化在CAVD中起着重要作用。因此，我们旨在研究雌激素相关受体α（ERR α）在hVIC成骨分化中的功能。我们发现，与正常对照组相比，CAVD样本中ERR α的水平显著升高。此外，ERR α在体外hVIC成骨分化过程中显著上调。进行功能获得和功能丧失实验以鉴定ERR α在体外hVIC钙化中的功能。内源性ERR α的抑制会减弱hVIC的钙化，而hVIC中ERR α的过表达会促进这一过程。RNA测序结果表明，血红素加氧酶-1（Hmox 1）是ERR α的下游靶点，这一点进一步通过蛋白质印迹得到证实。此外，我们还发现，通过shHmox 1下调Hmox 1有效地逆转了ERR α shRNA诱导的hVIC钙化抑制。ChIP-qPCR和荧光素酶分析表明，ERR α对Hmox 1有负调控作用。我们发现，Hmox 1或其底物的过表达显着抑制体外hVIC钙化。总之，我们发现ERR α的敲低可以通过上调Hmox 1来抑制hVIC钙化，并且ERR α和Hmox 1是治疗CAVD的潜在靶点。
Calcific aortic valve disease (CAVD) is the most common valvular heart disease in adults. The cellular mechanisms of CAVD are still unknown, but accumulating evidence has revealed that osteogenic differentiation of human valve interstitial cells (hVICs) plays an important role in CAVD. Thus, we aimed to investigate the function of estrogen-related receptor alpha (ERR alpha) in the osteogenic differentiation of hVICs. We found that the level of ERR alpha was significantly increased in CAVD samples compared to normal controls. In addition, ERR alpha was significantly upregulated during hVIC osteogenic differentiation in vitro. Gain- and loss-of-function experiments were performed to identify the function of ERR alpha in hVIC calcification in vitro. Inhibition of endogenous ERR alpha attenuated hVIC calcification, whereas overexpression of ERR alpha in hVICs promoted this process. RNA sequencing results suggested that heme oxygenase-1 (Hmox1) was a downstream target of ERR alpha, which was further confirmed by western blotting. Additionally, we also found that downregulation of Hmox1 by shHmox1 efficiently reversed the inhibition of calcification induced by ERR alpha shRNA in hVICs. ChIP-qPCR and luciferase assays indicated that Hmox1 was negatively regulated by ERR alpha. We found that overexpression of Hmox1 or its substrates significantly inhibited hVIC calcification in vitro. In conclusion, we found that knockdown of ERR alpha can inhibit hVIC calcification through upregulating Hmox1 and that ERR alpha and Hmox1 are potential targets for the treatment of CAVD.