Therapeutic Effect of Padina arborescens Extract on a Cell System Model for Parkinson’s Disease
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Reagent Treatments
2.2. Rat Primary Microglia and Astrocyte Preparation and Culture
2.3. In Vitro Kinase Assay
2.4. Western Blot Analysis
2.5. Messenger RNA (mRNA) Isolation and Complementary DNA (cDNA) Synthesis
2.6. Quantitative-Polymerase Chain Reaction (qPCR) Analysis
2.7. Enzyme-Linked Immunosorbent Assays (ELISAs) of Cytokines and Neurotropic Factors
2.8. Measurement of Cellular NO Levels
2.9. Detection of Cellular ROS
2.10. Seed-Accelerated Fibrilization of α-Synuclein
2.11. Thioflavin T Assay
2.12. Sandwich ELISA for α-Synuclein
2.13. Analysis of Lysosomal Activity
2.14. Cathepsin D Activity
2.15. Culturing of Worms
2.16. Lifespan Assay of C. elegans
2.17. Data Estimation and Statistical Analyses
3. Results
3.1. PA_EXT Inhibits LRRK2 Activity in BV2 Cells In Vitro
3.2. PA_EXT Decreases TNF-α Expression by Inhibiting LRRK2 Activity in BV2 Cells
3.3. PA_EXT Reduces the Gene Expression of TNF-α and iNOS in Rat Primary Microglia
3.4. PA_EXT Alleviates Oxidative Stress in Rat Primary Astrocytes
3.5. PA_EXT Blocks α-Synuclein Aggregation and Promotes Its Clearance in Differentiated SH-SY5Y (dSH) Cells
3.6. PA_EXT Prolongs the Lifespan of C. elegans Damaged by RTN
4. Discussion
5. Conclusions
6. Patents
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antibody | Manufacturer and Catalog Number |
---|---|
Anti-pS1292 LRRK2 | ab203181; Abcam, Cambridge, UK |
Anti-pS935 LRRK2 | ab133450; Abcam |
Anti-LRRK2 (N241A/34) | 75-253; NeuroMab, Davis, CA, USA |
Anti-β-actin | sc-47778; Santa Cruz, Dallas, TX, USA |
Anti-α-synuclein (Clone 42) | 610787; BD Bioscience, San Jose, CA, USA |
Peroxidase-conjugated AffiniPure goat anti-mouse IgG (H + L) | 115-035-003; Jackson Immunoresearch Laboratories, Inc., West Grove, PA, USA |
Peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H + L) | 115-035-144; Jackson Immunoresearch Laboratories, Inc. |
Genes | Sequence (5′-3′) | ||
---|---|---|---|
TNF-α | Rat | Forward | ACTGAACTTCGGGGTGATTG |
Reverse | GCTTGGTGGTTTGCTACGAC | ||
iNOS (NOS2) | Forward | CACCTTGGAGTTCACCCAGT | |
Reverse | ACCACTCGTACTTGGGATGC | ||
β-actin | Forward | CAGGGTGTGATGGTGGGTATGG | |
Reverse | AGTTGGTGACAATGCCGTGTTC | ||
TNF-α | Mouse | Forward | CCGATGGGTTGTACCTTGTC |
Reverse | GCTGGGTAGAGAATGGATGAACA | ||
β-actin | Forward | TGTTACCAACTGGGACGACA | |
Reverse | TCTCAGCTGTGGTGGTGAAG |
Group | Mean | S.E.M | N |
---|---|---|---|
Vehicle | 16.05 | 0.28 | 100 |
RTN | 9.38 | 0.47 | 100 |
RTN + PA_EXT | 9.7 | 0.4433 | 100 |
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Ho, D.H.; Kim, H.; Nam, D.; Seo, M.K.; Park, S.W.; Kim, D.-K.; Son, I. Therapeutic Effect of Padina arborescens Extract on a Cell System Model for Parkinson’s Disease. NeuroSci 2024, 5, 301-314. https://doi.org/10.3390/neurosci5030024
Ho DH, Kim H, Nam D, Seo MK, Park SW, Kim D-K, Son I. Therapeutic Effect of Padina arborescens Extract on a Cell System Model for Parkinson’s Disease. NeuroSci. 2024; 5(3):301-314. https://doi.org/10.3390/neurosci5030024
Chicago/Turabian StyleHo, Dong Hwan, Hyejung Kim, Daleum Nam, Mi Kyoung Seo, Sung Woo Park, Dong-Kyu Kim, and Ilhong Son. 2024. "Therapeutic Effect of Padina arborescens Extract on a Cell System Model for Parkinson’s Disease" NeuroSci 5, no. 3: 301-314. https://doi.org/10.3390/neurosci5030024