neuronal like cell line pc  (ATCC)

Journal: RSC Advances

Article Title: Comparative evaluation of donepezil-loaded polymeric and liposomal nanoparticles for Alzheimer's disease: biocompatibility, drug release kinetics, and cellular uptake study

doi: 10.1039/d6ra00929h

Figure Lengend Snippet: Effects of donepezil control solution, donepezil-loaded polymeric NPs, and donepezil-loaded DPPC-based liposomes treatment on ECV-304 and PC-12 cells. Cells were treated with increasing concentrations (3.125–50 µg mL −1 ) of each formulation in a time-dependent manner. All formulations maintained cell viability above 80% at concentrations up to 50 µg mL −1 in both cell lines, indicating acceptable biocompatibility. A Two-way ANOVA followed by Tukey's test was applied at each time point independently to compare the effect of different formulations across different concentrations within each cell line. **Significant with respect to the control ( p < 0.01), as shown by two-way ANOVA. The data is reported as mean ± SEM ( n = 3).

Article Snippet: The human endothelial-like cell line ECV-304 and the immortalized rat-derived neuronal-like cell line PC-12 were purchased from ATCC.

Techniques: Control, Liposomes, Formulation

Journal: RSC Advances

Article Title: Comparative evaluation of donepezil-loaded polymeric and liposomal nanoparticles for Alzheimer's disease: biocompatibility, drug release kinetics, and cellular uptake study

doi: 10.1039/d6ra00929h

Figure Lengend Snippet: In vitro cellular uptake of Rhodamine labelled DPPC-based liposomes (red) and FITC-labelled PCL/PVA nanoparticles (green) by ECV-304 and PC-12 cells after 24 hours of incubation period. Co-localization of the red fluorescence (liposomes) or green fluorescence (PCL/PVA NPs) in proximity to the blue DAPI-stained nuclei confirms the association of nanoparticles with ECV-304 and PC-12 cells, demonstrating either internalization or surface binding. Fluorescence microscopy was performed at 50× magnification; scale bar = 25 µm (consistent across all images).

Article Snippet: The human endothelial-like cell line ECV-304 and the immortalized rat-derived neuronal-like cell line PC-12 were purchased from ATCC.

Techniques: In Vitro, Liposomes, Incubation, Fluorescence, Staining, Binding Assay, Microscopy

Journal: RSC Advances

Article Title: Comparative evaluation of donepezil-loaded polymeric and liposomal nanoparticles for Alzheimer's disease: biocompatibility, drug release kinetics, and cellular uptake study

doi: 10.1039/d6ra00929h

Figure Lengend Snippet: Quantitative analysis of cellular uptake of rhodamine-labelled DPPC-based liposomes and FITC-labelled PCL/PVA NPs along with their respective free dye controls by ECV-304 and PC-12 cell lines. Both formulations showed significantly higher uptake than their respective free dye controls. PCL/PVA nanoparticles exhibited significantly higher uptake than liposomes in ECV-304 cells. Results are expressed as mean ± SD. Statistical significance was determined using a one-way ANOVA followed by Tukey's test to compare liposomes and nanoparticles uptake against their respective controls and denoted as follows: P < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

Article Snippet: The human endothelial-like cell line ECV-304 and the immortalized rat-derived neuronal-like cell line PC-12 were purchased from ATCC.

Techniques: Liposomes

Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling

doi: 10.1186/s12974-019-1406-7

Figure Lengend Snippet: Effects of Apelin 13 on the cytotoxicity and apoptosis induced by I/R in PC12 cells. a PC12 cells viability is assessed by measuring the CCK8 reduction. Results were shown as fold of control. b Effects of Apelin 13 on the levels of LDH in PC12 cell subjected to I/R. c Effects of Apelin 13 on PC12 cells apoptosis induced by I/R. PC12 cells were subjected to I/R with or without Apelin 13 pretreatment, then double stained with Annexin V/propidium iodide (PI). d Effects of Apelin 13 on the expression levels of Bax, Bcl-2, cleaved PARP, cleaved caspase-3, and cleaved caspase-9 in PC12 cell subjected to I/R. e ROS level is measured by a DHE kit using a laser confocal microscope. The effects of Apelin 13 on the levels of MDA ( f ), CAT ( g ), SOD ( h ), 8-Oxo-dG ( i ), IL-1( j ), TNF-α( k ), and COX-2 ( l ) were measured as described in the “ ” section. The columns and error bars were represented as means ± SD. ## P < 0.01 vs. the control group; * P < 0.05, ** P < 0.01 vs. the I/R treatment group

Article Snippet: The neuron-like rat pheochromocytoma cell line PC12 cells is obtained from American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM containing 10% fetal bovine serum and antibiotics (penicillin, 100 IU/ml; streptomycin, 100 μg/ml) at 37 °C in 5% CO 2 in a humidified incubator.

Techniques: Control, Staining, Expressing, Microscopy

Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling

doi: 10.1186/s12974-019-1406-7

Figure Lengend Snippet: Effects of Apelin 13 on expression of Nrf2-related antioxidant enzymes in PC12 cells subjected to I/R. a PC12 cells were pretreated with Apelin 13 for 6 h, then subjected to I/R. The expression levels of Nrf2, NQO-1, and HO-1 in cytoplasm were measured by western bolting as the method motioned in the “ ” section part. b The expression level of Nrf2 in nuclear is measured using the nuclear extract. c Effects of Apelin 13 on the activity of ARE determined by ARE luciferase report assay. d The cells were treated with Nrf2-specific siRNA (40 nM) or scrb siRNA for 48 h and Nrf2 expression level in cytoplasm was measured by western bolting. e Effects of siNrf2 on the protein expression levels of NQO-1, HO-1, and SOD. TNF-α ( f ), ROS ( g ), apoptosis rate ( h ), and cell viability ( i ) were measured after different treatments. The columns and error bars were represented as means ± SD. ## P < 0.01 vs. the control group; ** P < 0.01 vs. the I/R treatment group. && P < 0.05 vs. the scrb control RNA group

Article Snippet: The neuron-like rat pheochromocytoma cell line PC12 cells is obtained from American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM containing 10% fetal bovine serum and antibiotics (penicillin, 100 IU/ml; streptomycin, 100 μg/ml) at 37 °C in 5% CO 2 in a humidified incubator.

Techniques: Expressing, Western Blot, Activity Assay, Luciferase, Control

Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling

doi: 10.1186/s12974-019-1406-7

Figure Lengend Snippet: Effects of Apelin 13 were through AMPK and GSK-3β. a Apelin 13 increased the phosphorylation levels of AMPK and GSK-3β in a dose-dependent manner. b The cells were treated with AMPK-specific siRNA (40 nM) or scrb siRNA for 48 h, then treated with Apelin 13 and I/R, followed by western blotting. c Effects of siAMPK on the cell viability. d PC12 cells were transfected with the empty vector pcDNA3 or vectors encoding HA-tagged wild-type GSK-3β (WT-GSK3β-HA). Upon transfection, PC12 cells were exposed to different treatments as indicated. e Nrf2 nuclear expression levels after transfected with GSK-3β. f Effects of GSK-3β-HA on the cell viability. The columns and error bars were represented as means ± SD. ## P < 0.01 vs. the control group; ** P < 0.01 vs. the I/R treatment group. && P < 0.05 vs. the scrb control RNA group

Article Snippet: The neuron-like rat pheochromocytoma cell line PC12 cells is obtained from American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM containing 10% fetal bovine serum and antibiotics (penicillin, 100 IU/ml; streptomycin, 100 μg/ml) at 37 °C in 5% CO 2 in a humidified incubator.

Techniques: Phospho-proteomics, Western Blot, Transfection, Plasmid Preparation, Expressing, Control

Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling

doi: 10.1186/s12974-019-1406-7

Figure Lengend Snippet: The activation effects of Apelin 13 on AMPK is through AR/Gα/PLC/IP3/CaMKK. PC12 cells were pretreated with Apelin 13 with or without Gα q inhibitor Gp2A (10 μM) and Gα i inhibitor pertussis toxin (PTX) (200 ng/ml), then exposed to the indicated conditions. Cell viability ( a ) and ROS levels ( b ) were measured using relative kit. c Gp2A and PTX abolished the effects of Apelin 13 on the activation of AMPK/GSK-3β/Nrf2 pathway. d Apelin 13 induced the expression of PLC, IP3, and CaMKK in a dose-dependent manner. e PC12 cells were pretreated with Apelin 13 with or without CaMKK inhibitor STO-609 (1 μg/ml) and IP3 inhibitor 2-APB (1 μg/ml), then exposed to the indicated conditions. The phosphorylation of AMPK and GSK-3β ( e ) and the expression of CaMKK ( f ) were determined by western blotting. g PC12 cells were pretreated with Apelin 13 with or without apelin receptor inhibitor F13A (1 μM), then exposed to the indicated conditions. F13A inhibited the expression of PLC, IP3, CaMKK ( g ) and the phosphorylation of AMPK and GSK-3β ( h ) and increased the ROS levels ( i ) in PC12 cells. The columns and error bars were represented as means ± SD. ## P < 0.01 vs. the control group; ** P < 0.01 vs. the I/R treatment group. && P < 0.05 vs. the Apelin 13 treatment group

Article Snippet: The neuron-like rat pheochromocytoma cell line PC12 cells is obtained from American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM containing 10% fetal bovine serum and antibiotics (penicillin, 100 IU/ml; streptomycin, 100 μg/ml) at 37 °C in 5% CO 2 in a humidified incubator.

Techniques: Activation Assay, Expressing, Phospho-proteomics, Western Blot, Control

Journal: Journal of Neuroinflammation

Article Title: Neuroprotective effect of Apelin 13 on ischemic stroke by activating AMPK/GSK-3β/Nrf2 signaling

doi: 10.1186/s12974-019-1406-7

Figure Lengend Snippet: AR/Gα/PLC/IP3/CaMKK is involved in the activation of AMPK/GSK-3β/Nrf2 pathway in vivo . a The effects of Apelin 13 on the expression of CaMKK and IP3 in the brain. b In ARKO rats, the AMPK/GSK-3β/Nrf2 pathway was inhibited and the expression levels of PLC, IP3, CaMKK, and Nrf2 were also decreased. The levels of SOD ( c ), ROS ( d ), infarct volume ratio ( e ), caspase 3 ( f ) IL-1β ( g ), and TNF-α ( h ) were measured as before. The columns and error bars were represented as means ± SD. ## P < 0.01 vs. the sham group; ** P < 0.01 vs. the I/R treatment group. && P < 0.05 vs. the WT rats. i Potential mechanism underlying the neuro-protective effects of Apelin 13 on I/R-induced brain injuries in rats and PC12 cells. Apelin 13 protected the brain against ischemic stroke-induced oxidative stress and inflammation through AMPK-mediated inhibitory phosphorylation of GSK-3β downstream of AR/G-coupled receptors pathway, and further induced Nrf2-mediated antioxidant proteins expressions

Article Snippet: The neuron-like rat pheochromocytoma cell line PC12 cells is obtained from American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM containing 10% fetal bovine serum and antibiotics (penicillin, 100 IU/ml; streptomycin, 100 μg/ml) at 37 °C in 5% CO 2 in a humidified incubator.

Techniques: Activation Assay, In Vivo, Expressing, Phospho-proteomics