Abstract
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.
Highlights
In the last two decades, attention to nanomaterials, especially metal nanomaterials, has grown significantly due to their interesting and unique p roperties[1,2,3,4,5]
About the neuroprotective properties of CuNPs green-synthesized by plants, the neuroprotective effects of Achillea biebersteinii leaf aqueous extract synthesized CuNPs against methamphetamine-induced cell death in PC12 were investigated by Wang et al (2020)
Copper nanoparticles were prepared and synthesized in a watered environment using C. sativus leaf extract as decreasing and stabilizing agents. We investigated their potential against methadone-induced cell death in PC12
Summary
Crocus sativus L. leaf aqueous extract on methadone‐induced cell death in adrenal phaeochromocytoma (PC12) cell line. 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. Based on the neuroprotective properties of C. sativus and CuNPs, we have attempted to determine the CuNPs effects comprising C. sativus leaf aqueous extract on cellular and molecular variables like viability and cell death index in methadone-induced PC12 cell death In these cells, the activity of caspase-3, inflammatory cytokine concentration, and mitochondrial membrane potential were examined to effectively figure out the functioning mechanism of the CuNPs. Experimental Material. For Cu(NO3)[2], C. sativus leaf aqueous extract, CuNPs solutions in the RPMI1640 water cell cultural environment were first dissolved in DMSO and added to the cultural environment with a final volume of 0.1%. The data obtained at the end of the study were fed to the SPSS-22 program and the Tukey post-hoc test (p ≤ 0.01) after the one-sided ANOVA test
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