Abstract
Purpose: To investigate the neuroprotective activity of the aqueous extract of Sutherlandia frutescens (SF) against 1-methyl-4-phenylpyridinium (MPP+)-induced toxicity in SH-SY5Y neuroblastoma cells.
 Methods: SH-SY5Y neuroblastoma cells were divided into different treatment groups: untreated cells, cells treated with MPP+ alone (2 mM), cells pretreated with SF (20 μg) prior to MPP+ (2 mM) treatment and cells treated with SF (20 μg) alone. Twenty-four hours after treatment with MPP+, cell viability was assessed by MTT assay, and changes in cell morphology, intracellular reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP) as well as caspases 3/7 and 9 activities were determined.
 Results: Treatment of SH-SY5Y cells with MPP+ alone significantly altered cellular morphology, increased ROS production (p = 0.005), induced a significant loss of MMP (p = 0.0011) and caused significant apoptotic cell death, via the activation of caspases 3/7 and 9 (p ≤ 0.0359). These effects were however significantly (p ≤ 0.0359) attenuated in cells pre-treated with the aqueous leaf extract of SF, indicating the possible neuroprotective activity of the SF extract.
 Conclusion: The results of this study suggest that the aqueous leaf extract of SF may be neuroprotective against MPP+-induced toxicity via apoptotic cell death and inhibition of ROS production. Further mechanistic studies are required to validate the results of the present study using additional PD models, different extract preparations and active compounds derived from SF.
 Keywords: Parkinson’s disease, MPP+, Sutherlandia frutescens, Reactive oxygen species, Apoptosis, Neurodegeneration
Highlights
Parkinson’s disease (PD) is the second most common, age-dependent neurodegenerative disorder resulting from continuous loss of dopaminergic neurons in the pars compacta of the substantia nigra of the midbrain (SNpc)
P < 0.0005 vs untreated SH-SY5Y cells; *p < 0.05 vs SH-SY5Y cells treated with MPP+ only
The MTT assay results (Figure 1C) showed that treatment with MPP+ alone caused a significant decrease in cell viability (p < 0.0001), while pretreatment with Sutherlandia frutescens (SF) before exposure to MPP+ resulted in significant increase (p = 0.0008,) in cell viability
Summary
Parkinson’s disease (PD) is the second most common, age-dependent neurodegenerative disorder resulting from continuous loss of dopaminergic neurons in the pars compacta of the substantia nigra of the midbrain (SNpc). This study investigated the mechanisms by which SF could potentially provide neuroprotection against MPP+-induced cell death in the SH-SY5Y human neuroblastoma cell line used as a model of PD. The TB dye only enters non-viable cells through their damaged cell membranes while healthy viable cells exclude this dye For this assay, the SH-SY5Y cells were seeded at a density of 1.1 x 105 /ml in 60 mm dishes and treated as previously described. The SH-SY5Y cells were seeded at a density of 1.2 x 105 /mL in 100 mm dishes and treated as previously described. SH-SY5Y cells were seeded at a density of 1.2 x 105 cells /mL in 100 mm dishes and treated as earlier described. SH-SY5Y cells were seeded at a density of 1.0 x 104 cells/well in white-walled 96well plates and treated as earlier described.
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