Blocking of P2X7r Reduces Mitochondrial Stress Induced by Alcohol and Electronic Cigarette Exposure in Brain Microvascular Endothelial Cells.
Blocking of P2X7r Reduces Mitochondrial Stress Induced by Alcohol and Electronic Cigarette Exposure in Brain Microvascular Endothelial Cells.
复制标题
DOI：
10.3390/antiox11071328
复制
发表时间：
2022-07-06
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
在人类和动物模型中的研究表明，长期酒精/电子烟（e-Cig）暴露会影响线粒体功能，并损害脑微血管内皮细胞（BMVEC）的屏障功能。识别慢性酒精/电子烟暴露诱导BMVEC中线粒体损伤的信号通路对于保护血脑屏障（BBB）至关重要。为了解决这个问题，我们用乙醇（ETH）[100 mM]、乙醛（ALD）[100 μM]或电子烟（e-Cig）[35 ng/mL的1.8%或0%尼古丁]条件培养基处理人BMVEC [hBMVEC（D3细胞系）]，并通过Seahorse分析仪测量显示线粒体氧化磷酸化（OXPHOS）降低。海马数据进一步补充了线粒体OXPHOS蛋白的表达检测蛋白质印迹。我们还观察到由于ETH、ALD或1. 8% e-Cig暴露引起的线粒体膜电位的破坏，ATP的胞质逃逸及其细胞外释放。此外，使用qPCR测量，ETH、ALD或1. 8% e-Cig处理导致嘌呤能P2 X7 r和TRPV 1通道基因表达升高。我们还证明了P2 X7 r拮抗剂A804598（10 μM）在恢复线粒体氧化磷酸化水平和阻止细胞外ATP释放方面的保护作用。在使用跨内皮电阻的BBB功能测定中，我们表明阻断P2 X7 r通道增强屏障功能。总之，我们确定了由ETH，ALD和1.8%电子烟引起的线粒体损伤的潜在共同途径，这些途径允许新的保护性干预措施。我们正在进一步研究P2 X7调控途径与线粒体健康之间的潜在联系。
Studies in both humans and animal models demonstrated that chronic alcohol/e-cigarette (e-Cig) exposure affects mitochondrial function and impairs barrier function in brain microvascular endothelial cells (BMVECs). Identification of the signaling pathways by which chronic alcohol/e-Cig exposure induces mitochondrial damage in BMVEC is vital for protection of the blood–brain barrier (BBB). To address the issue, we treated human BMVEC [hBMVECs (D3 cell-line)] with ethanol (ETH) [100 mM], acetaldehyde (ALD) [100 μM], or e-cigarette (e-Cig) [35 ng/mL of 1.8% or 0% nicotine] conditioned medium and showed reduced mitochondrial oxidative phosphorylation (OXPHOS) measured by a Seahorse analyzer. Seahorse data were further complemented with the expression of mitochondrial OXPHOS proteins detected by Western blots. We also observed cytosolic escape of ATP and its extracellular release due to the disruption of mitochondrial membrane potential caused by ETH, ALD, or 1.8% e-Cig exposure. Moreover ETH, ALD, or 1.8% e-Cig treatment resulted in elevated purinergic P2X7r and TRPV1 channel gene expression, measured using qPCR. We also demonstrated the protective role of P2X7r antagonist A804598 (10 μM) in restoring mitochondrial oxidative phosphorylation levels and preventing extracellular ATP release. In a BBB functional assay using trans-endothelial electrical resistance, we showed that blocking the P2X7r channel enhanced barrier function. In summary, we identified the potential common pathways of mitochondrial injury caused by ETH, ALD, and 1.8% e-Cig which allow new protective interventions. We are further investigating the potential link between P2X7 regulatory pathways and mitochondrial health.
登录
查看更多内容
影响因子：
33.9
作者：
Gentzke, Andrea S.;Creamer, MeLisa;King, Brian A.
通讯作者：
King, Brian A.
炎症过程中的核苷酸信号传导。
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
5.6
作者：
Burnstock G
通讯作者：
Burnstock G
影响因子：
5.6
作者：
Dosch M;Gerber J;Jebbawi F;Beldi G
通讯作者：
Beldi G
影响因子：
11.4
作者：
Kaisar MA;Villalba H;Prasad S;Liles T;Sifat AE;Sajja RK;Abbruscato TJ;Cucullo L
通讯作者：
Cucullo L