Cannabidiol Promotes Endothelial Cell Survival by Heme Oxygenase-1-Mediated Autophagy
Cannabidiol Promotes Endothelial Cell Survival by Heme Oxygenase-1-Mediated Autophagy
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DOI：
10.3390/cells9071703
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发表时间：
2020-07-01
期刊：
影响因子：
6
通讯作者：
Hinz, Burkhard
中科院分区：
文献类型：
--
作者：
Boeckmann, Sabine;Hinz, Burkhard
大麻二酚（CBD），一种非精神活性大麻素，已被报道在内皮细胞中介导抗氧化、抗炎和抗血管生成作用。本研究探讨CBD对血红素氧合酶-1（HO-1）表达的影响及其在调节人脐静脉内皮细胞（HUVEC）代谢、自噬和凋亡过程中的功能作用。浓度高达10 μ M CBD显示HO-1 mRNA和蛋白质的浓度依赖性增加和HO-1调节转录因子核因子红细胞2相关因子2（Nrf 2）的增加。CBD诱导的HO-1的表达并没有被大麻素激活受体（CB 1，CB 2，瞬时受体电位香草素1）的拮抗剂降低，但被活性氧（ROS）清除剂N-乙酰-L-半胱氨酸（NAC）。HUVEC与6 μ M CBD的孵育导致代谢活性增加，而10 μ M CBD导致代谢活性降低和诱导凋亡，如通过增强的caspase-3切割所证明的。此外，CBD引发了自噬标志物LC 3A/B-II的浓度依赖性增加。CBD诱导的LC 3A/B-II水平和半胱天冬酶-3裂解均被NAC降低。巴弗洛霉素A对自噬的抑制（1）导致6 μ M CBD诱导凋亡，10 μ M CBD的促凋亡作用进一步增加。另一方面，用锡原卟啉IX（SnPPIX）抑制HO-1活性或用Nrf 2 siRNA敲低HO-1表达与CBD介导的自噬和细胞凋亡的减少有关。总之，我们的数据首次显示，ROS介导的HO-1在内皮细胞中的表达是CBD介导保护性自噬的机制，然而，在较高的CBD浓度下，它不再能阻止诱导细胞凋亡的细胞死亡。
Cannabidiol (CBD), a non-psychoactive cannabinoid, has been reported to mediate antioxidant, anti-inflammatory, and anti-angiogenic effects in endothelial cells. This study investigated the influence of CBD on the expression of heme oxygenase-1 (HO-1) and its functional role in regulating metabolic, autophagic, and apoptotic processes of human umbilical vein endothelial cells (HUVEC). Concentrations up to 10 mu M CBD showed a concentration-dependent increase of HO-1 mRNA and protein and an increase of the HO-1-regulating transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2). CBD-induced HO-1 expression was not decreased by antagonists of cannabinoid-activated receptors (CB1, CB2, transient receptor potential vanilloid 1), but by the reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine (NAC). The incubation of HUVEC with 6 mu M CBD resulted in increased metabolic activity, while 10 mu M CBD caused decreased metabolic activity and an induction of apoptosis, as demonstrated by enhanced caspase-3 cleavage. In addition, CBD triggered a concentration-dependent increase of the autophagy marker LC3A/B-II. Both CBD-induced LC3A/B-II levels and caspase-3 cleavage were reduced by NAC. The inhibition of autophagy by bafilomycin A(1)led to apoptosis induction by 6 mu M CBD and a further increase of the proapoptotic effect of 10 mu M CBD. On the other hand, the inhibition of HO-1 activity with tin protoporphyrin IX (SnPPIX) or knockdown of HO-1 expression by Nrf2 siRNA was associated with a decrease in CBD-mediated autophagy and apoptosis. In summary, our data show for the first time ROS-mediated HO-1 expression in endothelial cells as a mechanism by which CBD mediates protective autophagy, which at higher CBD concentrations, however, can no longer prevent cell death inducing apoptosis.