Laboratory of Molecular Neurobiology, Academy of Biology and Biotechnology, Southern Federal University, pr. Stachki 194/1, Rostov-on-Don, 344090, Russia.
Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London, WC1N 3BG, UK.
Mol Neurobiol. 2018 Jan;55(1):96-102. doi: 10.1007/s12035-017-0721-1.
Photodynamic therapy (PDT) leads to production of reactive oxygen species (ROS) and cell destruction due to oxidative stress. We used photodynamic effect of photosensitizer radachlorin to unravel the effect of photo-induced oxidative stress on the calcium signal and lipid peroxidation in primary culture of cortical neurons and astrocytes using live cell imaging. We have found that irradiation in presence of 200 nM of radachlorin induces calcium signal in primary neurons and astrocytes. Photo-induced neuronal calcium signal depends on internal calcium stores as it was still observed in calcium-free medium and could be blocked by depletion of endoplasmic reticulum (ER) stores with inhibitor of sarco-endoplasmic reticulum Ca ATPase (SERCA) thapsigargin. Both inhibitors of phospholipase C activity U73122 and water-soluble analogue of vitamin E Trolox suppressed calcium response activated by PDT. We have also observed that the photodynamic effect of radachlorin induces lipid peroxidation in neurons and astrocytes. This data demonstrate that lipid peroxidation induced by PDT in neurons and astrocytes leads to activation of phospholipase C that results in production of inositol 1,4,5-trisphosphate (IP3).
光动力疗法（PDT）会导致活性氧（ROS）的产生和细胞破坏，这是由于氧化应激引起的。我们使用光敏剂拉达霉素的光动力效应，使用活细胞成像技术，在皮质神经元和星形胶质细胞的原代培养物中研究光诱导氧化应激对钙信号和脂质过氧化的影响。我们发现，在 200 nM 拉达霉素存在的情况下照射会诱导原代神经元和星形胶质细胞产生钙信号。光诱导的神经元钙信号依赖于细胞内钙库，因为在无钙培养基中仍然可以观察到这种信号，并且可以通过内质网（ER）储存抑制剂肌浆内质网 Ca ATP 酶（SERCA） thapsigargin 耗尽来阻断。两种 PLC 活性抑制剂 U73122 和维生素 E 水溶性类似物 Trolox 均可抑制 PDT 激活的钙反应。我们还观察到，拉达霉素的光动力效应会在神经元和星形胶质细胞中诱导脂质过氧化。这些数据表明，PDT 在神经元和星形胶质细胞中诱导的脂质过氧化会导致 PLC 的激活，从而产生肌醇 1,4,5-三磷酸（IP3）。
