Qiao Hui, Li Yun, Xu Zhendong, Li Wenxian, Fu Zhijian, Wang Yuezhi, King Alexander, Wei Huafeng
From the Department of Anesthesiology and Critical Care, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania (H.Q., Y.L., Z.X., A.K., H.W.); Department of Anesthesiology, The Eye Ear Nose and Throat Hospital of Fudan University, Shanghai, People's Republic of China (H.Q., W.L.); Department of Pain Medicine, Provincial Hospital Affiliated with Shandong University, Jinan, People's Republic of China (Y.L., Z.F.); Department of Anesthesiology, First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, People's Republic of China (Z.X.); and Department of Gerontology, Huashan Hospital of Fudan University, Shanghai, People's Republic of China (Y.W.).
Anesthesiology. 2017 Sep;127(3):490-501. doi: 10.1097/ALN.0000000000001730.
In human cortical neural progenitor cells, we investigated the effects of propofol on calcium homeostasis in both the ryanodine and inositol 1,4,5-trisphosphate calcium release channels. We also studied propofol-mediated effects on autophagy, cell survival, and neuro- and gliogenesis.
The dose-response relationship between propofol concentration and duration was studied in neural progenitor cells. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase release assays. The effects of propofol on cytosolic calcium concentration were evaluated using Fura-2, and autophagy activity was determined by LC3II expression levels with Western blot. Proliferation and differentiation were evaluated by bromodeoxyuridine incorporation and immunostaining with neuronal and glial markers.
Propofol dose- and time-dependently induced cell damage and elevated LC3II expression, most robustly at 200 µM for 24 h (67 ± 11% of control, n = 12 to 19) and 6 h (2.4 ± 0.5 compared with 0.6 ± 0.1 of control, n = 7), respectively. Treatment with 200 μM propofol also increased cytosolic calcium concentration (346 ± 71% of control, n = 22 to 34). Propofol at 10 µM stimulated neural progenitor cell proliferation and promoted neuronal cell fate, whereas propofol at 200 µM impaired neuronal proliferation and promoted glial cell fate (n = 12 to 20). Cotreatment with ryanodine and inositol 1,4,5-trisphosphate receptor antagonists and inhibitors, cytosolic Ca chelators, or autophagy inhibitors mostly mitigated the propofol-mediated effects on survival, proliferation, and differentiation.
These results suggest that propofol-mediated cell survival or neurogenesis is closely associated with propofol's effects on autophagy by activation of ryanodine and inositol 1,4,5-trisphosphate receptors.
在人皮质神经祖细胞中，我们研究了丙泊酚对兰尼碱和肌醇1,4,5 -三磷酸钙释放通道钙稳态的影响。我们还研究了丙泊酚介导的对自噬、细胞存活以及神经发生和胶质发生的影响。
在神经祖细胞中研究了丙泊酚浓度与作用持续时间之间的剂量反应关系。通过3 -（4,5 -二甲基噻唑 - 2 -基）- 2,5 -二苯基四氮唑溴盐和乳酸脱氢酶释放试验测定细胞活力。使用Fura - 2评估丙泊酚对胞质钙浓度的影响，通过蛋白质印迹法检测LC3II表达水平来确定自噬活性。通过溴脱氧尿苷掺入以及用神经元和胶质细胞标志物进行免疫染色来评估增殖和分化。
丙泊酚剂量和时间依赖性地诱导细胞损伤并提高LC3II表达，在200 μM作用24小时（为对照组的67±11%，n = 12至19）和6小时（与对照组的0.6±0.1相比为2.4±0.5，n = 7）时最为显著。用200 μM丙泊酚处理也增加了胞质钙浓度（为对照组的346±71%，n = 22至34）。10 μM的丙泊酚刺激神经祖细胞增殖并促进神经元细胞命运，而200 μM的丙泊酚损害神经元增殖并促进胶质细胞命运（n = 12至20）。与兰尼碱和肌醇1,4,5 -三磷酸受体拮抗剂及抑制剂、胞质钙螯合剂或自噬抑制剂共同处理大多减轻了丙泊酚介导的对存活、增殖和分化的影响。
这些结果表明，丙泊酚介导的细胞存活或神经发生与丙泊酚通过激活兰尼碱和肌醇1,4,5 -三磷酸受体对自噬的影响密切相关。
