Rifampicin Prevents SH-SY5Y Cells from Rotenone-Induced Apoptosis via the PI3K/Akt/GSK-3β/CREB Signaling Pathway

Abstract

In addition to its original application for treating tuberculosis, rifampicin has multiple potential neuroprotective effects in chronic neurodegenerative diseases including Parkinson's disease (PD) and Alzheimer's disease. Inflammatory reactions and the PI3K/Akt pathway are strongly implicated in dopaminergic neuronal death in PD. This study aims to investigate whether rifampicin protects rotenone-lesioned SH-SY5Y cells via regulating PI3K/Akt/GSK-3β/CREB pathway. Rotenone-treated SH-SY5Y cells were used as the cell model to investigate the neuroprotective effects of rifampicin. Cell viability and apoptosis of SH-SY5Y cells were determined by CCK-8 assay and flow cytometry, respectively. The expression of Akt, p-Akt, GSK-3β, p-GSK-3β, CREB and p-CREB were measured by Western blot. Our results showed that the cell viability and level of phospho-CREB significantly decreased in SH-SY5Y cells exposed to rotenone when compared to the control group. Both the cell viability and the expression of phospho-CREB in cells pretreated with rifampicin were higher than those of cells exposed to rotenone alone. Moreover, pretreatment of SH-SY5Y cells with rifampicin enhanced phosphorylation of Akt and suppressed activity of GSK-3β. The addition of LY294002, a PI3K inhibitor, could suppress phosphorylation of Akt and CREB and activate GSK-3β, resulting in abolishment of neuroprotective effects of rifampicin on cells exposed to rotenone. Rifampicin provides neuroprotection against dopaminergic degeneration, partially via the PI3K/Akt/GSK-3β/CREB signaling pathway. These findings suggest that rifampicin could be an effective and promising neuroprotective candidate for treating PD.

Caspase-4 mediates cytoplasmic accumulation of TDP-43 in the primate brains

“…Tunicamycin has been reported to increase caspase-4 activity [ 18 , 58 ] and reduce the viability of SH-SY5Y cells [ 16 , 61 ]. We therefore measured cell viability of the cultured SH-SY5Y cells using CCK-8, a widely used assay that can sensitively measure the viability of a variety of cultured cells including SH-SY5Y cells [ 10 , 56 ]. We found that inhibition of caspase-4 via siRNA-caspase-4 also significantly increased cell viability after tunicamycin treatment in parallel with reduction of cytoplasmic accumulation of endogenous TDP-43 (Fig.…”

“…It utilizes tetrazolium-8-[2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium] monosodium salt that produces a water-soluble formazan dye in living cells. The amount of this dye is directly proportional to the number of living cells, offering a sensitive assay to detect viability of many cell lines including SH-SY5Y cells [ 10 , 56 ]. Briefly, SH-SY5Y cells were transfected with siRNA for 48 h and then treated with tunicamycin (1 μg/ml) for 12 h. An equal number of 100 µl SH-SY5Y cell suspension (1 × 10 4 cells/100 µl/well) was dispensed in a 96-well plate.…”

In Vitro and In Vivo Neuroprotective Effects of Stellettin B Through Anti-Apoptosis and the Nrf2/HO-1 Pathway

“…SB exerts its neuroprotective activity via both anti-apoptotic and anti-oxidative stress pathways, and in this regard, PI3K/Akt, ERK, and P38 have been widely discussed as relevant anti-apoptotic pathways in previous PD-related research. The PI3K/Akt pathways play a crucial role in dopaminergic cell survival in PD [27,45], and, therefore, molecules such as rifampicin and resveratrol that act via PI3K/Akt pathways have been investigated as potential therapeutic agents in in vitro models of PD [46,47,48]. Wu et al reported that treatment with 10 μM rifampicin could significantly inhibit rotenone-induced cell apoptosis via activation of the PI3K/Akt pathway [48], and systemic treatment with rifampicin was neuroprotective in a murine PD model [49].…”

Hydrogen exerts neuroprotection by activation of the miR‐21/PI3K/AKT/GSK‐3β pathway in an in vitro model of traumatic brain injury

“…Therefore, GSK‐3β is considered a therapeutic target for a variety of nervous system diseases. Some studies using in vivo or in vitro models indicate that GSK‐3β may regulate neuronal apoptosis through activation of NF‐κB, β‐catenin or CREB signalling pathways . Other studies report that GSK‐3β can activate Nrf2 signalling to attenuate β‐amyloid‐induced oxidative stress in Alzheimer's disease models .…”