Department of Neurosurgery, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, Medical College of Henan University, Zhengzhou, 450003, Henan, China.
Department of Neurosurgery, Xi'an No. 1 Hospital, No. 30 South Street Powder Lane, Beilin District, Xi'an, 710002, Shaanxi, China.
Sci Rep. 2020 Jul 15;10(1):11631. doi: 10.1038/s41598-020-68142-8.
In this research, we prepared and formulated a neuroprotective supplement (copper nanoparticles in aqueous medium utilizing Crocus sativus L. Leaf aqueous extract) for determining its potential against methadone-induced cell death in PC12. The results of chemical characterization tests i.e., FE-SEM, FT-IR, XRD, EDX, TEM, and UV-Vis spectroscopy revealed that the study showed that copper nanoparticles were synthesized in the perfect way possible. In the TEM and FE-SEM images, the copper nanoparticles were in the mean size of 27.5 nm with the spherical shape. In the biological part of the present research, the Rat inflammatory cytokine assay kit was used to measure the concentrations of inflammatory cytokines. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) test was used to show DNA fragmentation and apoptosis. Caspase-3 activity was assessed by the caspase activity colorimetric assay kit and mitochondrial membrane potential was studied by Rhodamine123 fluorescence dye. Also, the cell viability of PC12 was measured by trypan blue assay. Copper nanoparticles-treated cell cutlers significantly (p ≤ 0.01) decreased the inflammatory cytokines concentrations, caspase-3 activity, and DNA fragmentation and they raised the cell viability and mitochondrial membrane potential in the high concentration of methadone-treated PC12 cells. The best result of neuroprotective properties was seen in the high dose of copper nanoparticles i.e., 4 µg. According to the above results, copper nanoparticles containing C. sativus leaf aqueous extract can be used in peripheral nervous system treatment as a neuroprotective promoter and central nervous system after approving in the clinical trial studies in humans.
在这项研究中，我们制备并配方了一种神经保护补充剂（铜纳米粒子在含有藏红花叶水提物的水介质中），以确定其对甲氨蝶呤诱导的 PC12 细胞死亡的潜在作用。化学特性测试的结果，即 FE-SEM、FT-IR、XRD、EDX、TEM 和 UV-Vis 光谱表明，研究表明铜纳米粒子以可能的最佳方式合成。在 TEM 和 FE-SEM 图像中，铜纳米粒子的平均尺寸为 27.5nm，呈球形。在本研究的生物学部分，使用大鼠炎症细胞因子试剂盒测量炎症细胞因子的浓度。末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）试验用于显示 DNA 片段化和细胞凋亡。通过 caspase 活性比色法试剂盒评估 caspase-3 活性，通过 Rhodamine123 荧光染料研究线粒体膜电位。此外，通过台盼蓝测定法测量 PC12 的细胞活力。铜纳米粒子处理的细胞碎片显著（p≤0.01）降低了炎症细胞因子浓度、caspase-3 活性和 DNA 片段化，并且提高了高浓度甲氨蝶呤处理的 PC12 细胞中的细胞活力和线粒体膜电位。铜纳米粒子含有藏红花叶水提物的最佳神经保护特性出现在高剂量（4μg）。根据上述结果，含有藏红花叶水提物的铜纳米粒子可作为神经保护促进剂用于外周神经系统治疗，并可在人体临床试验中得到批准后用于中枢神经系统。
