Neural-Induced Human Adipose Tissue-Derived Stem Cells Conditioned Medium Ameliorates Rotenone-Induced Toxicity in SH-SY5Y Cells

Abstract

Parkinson’s disease (PD) is an age-related neurodegenerative disease (NDD) characterized by the degenerative loss of dopaminergic neurons in the substantia nigra along with aggregation of α-synuclein (α-syn). Neurogenic differentiation of human adipose-derived stem cells (NI-hADSCs) by supplementary factors for 14 days activates different biological signaling pathways. In this study, we evaluated the therapeutic role of NI-hADSC-conditioned medium (NI-hADSC-CM) in rotenone (ROT)-induced toxicity in SH-SY5Y cells. Increasing concentrations of ROT led to decreased cell survival at 24 and 48 h in a dose- and time-dependent manner. Treatment of NI-hADSC-CM (50% dilution in DMEM) against ROT (0.5 μM) significantly increased the cell survival. ROT toxicity decreased the expression of tyrosine hydroxylase (TH). Western blot analysis of the Triton X-100-soluble fraction revealed that ROT significantly decreased the oligomeric, dimeric, and monomeric phosphorylated Serine129 (p-S129) α-syn, as well as the total monomeric α-syn expression levels. ROT toxicity increased the oligomeric, but decreased the dimeric and monomeric p-S129 α-syn expression levels. Total α-syn expression (in all forms) was increased in the Triton X-100-insoluble fraction, compared to the control. NI-hADSC-CM treatment enhanced the TH expression, stabilized α-syn monomers, reduced the levels of toxic insoluble p-S129 α-syn, improved the expression of neuronal functional proteins, regulated the Bax/Bcl-2 ratio, and upregulated the expression of pro-caspases, along with PARP-1 inactivation. Moreover, hADSC-CM treatment decreased the cell numbers and have no effect against ROT toxicity on SH-SY5Y cells. The therapeutic effects of NI-hADSC-CM was higher than the beneficial effects of hADSC-CM on cellular signaling. From these results, we conclude that NI-hADSC-CM exerts neuroregenerative effects on ROT-induced PD-like impairments in SH-SY5Y cells.

Neuroprotective effects of a Wnt antagonist in quinolinic acid-induced excitotoxicity in N18D3 cells

“…The cells were cultured and prepared for a western blot assay following our previous methods (15)(16)(17)(18). Wnt exposed to 30 mM QUIN, the cell survival rate was 43.5 ± 2.0055% (**p < 0.01).…”

Neuroprotective Effects of the Neural-Induced Adipose-Derived Stem Cell Secretome against Rotenone-Induced Mitochondrial and Endoplasmic Reticulum Dysfunction

“…Rotenone (ROT), a lipophilic piscicidal compound isolated from the roots of the subtropical plant species of Lonchocarpus and Derris suppresses the flow of electrons from the iron–sulfur centers in mitochondrial electron transport chain complex I. ROT reproduces PD-like impairments, such as decreased tyrosine hydroxylase, increased phosphorylation and aggregation of α-syn, and imbalanced autophagy degradation, which induces apoptotic death in SH-SY5Y neuroblastoma cells [ 15 , 16 ]. Treatments for PD mainly focus on restoring mitochondrial function and subsequently relieving motor symptoms, such as tremors, bradykinesia, and rigidity.…”

Systemic Rotenone Administration Causes Extra-Nigral Alterations in C57BL/6 Mice

“…Rotenone administered at low dosages induces degeneration of the dopaminergic nigrostriatal pathway and causes motor deficits [ 12 , 13 , 14 , 15 , 16 ]. Rotenone-mediated dopaminergic degeneration occurs via the inhibition of the mitochondria electron transport chain complex I, which results in the formation and accumulation of reactive oxygen species, leading to oxidative stress [ 9 , 17 ]. Studies of human post-mortem brain tissues indicate that oxidative stress and accumulation of reactive oxygen species are critical events in the pathogenesis of PD [ 18 , 19 ].…”