Synergic pro-apoptotic effects of Ferulic Acid and nanostructured lipid carrier in glioblastoma cells assessed through molecular and Delayed Luminescence studies

Abstract

Herein, we assessed the effect of Ferulic Acid (FA), a natural antioxidant with anti-cancer effect, on the human glioblastoma cells through molecular and Delayed Luminescence (DL) studies. DL, a phenomenon of ultra-week emission of optical photons, was used to monitor mitochondrial assessment. The effect of FA loaded in nanostructured lipid carriers (NLCs) was also assessed. To validate NLCs as a drug delivery system for glioblastoma treatment, particular attention was focused on their effect. We found that free FA induced a significant decrease in c-Myc and Bcl-2 expression levels accompanied by the apoptotic pathway activation. Blank NLCs, even if they did not induce cytotoxicity and caspase-3 cleavage, decreased Bcl-2, ERK1/2, c-Myc expression levels activating PARP-1 cleavage. The changes in DL intensity and kinetics highlighted a possible effect of nanoparticle matrix on mitochondria, through the involvement of the NADH pool and ROS production that, in turn, activates ERK1/2 pathways. All the effects on protein expression levels and on the activation of apoptotic pathway appeared more evident when the cells were exposed to FA loaded in NLCs. We demonstrated that the observed effects are due to a synergic pro-apoptotic influence exerted by FA, whose bioavailability increases in the glioblastoma cells, and NLCs formulation.Glioblastoma multiforme (GBM), also known as grade IV astrocytoma, represents the most prevalent and aggressive brain cancer. It is characterized by glial cells and has finger-like tentacles that infiltrate the brain, which make them very difficult to remove with surgical procedures. GBM exhibits a high level of resistance to conventional chemotherapy and radiotherapy, also due to the existence of blood-brain barrier (BBB), glioma stem cells and complex network of multiple modified signalling pathways 1 . The most frequent aberrant expression is represented by the dysregulation of extracellular signal-regulated protein kinase (ERK), which is associated with poor survival of the patients. The ERK isoforms (p42/44 or ERK1/ERK2) by interacting with specific phosphorylation substrates, play a pivotal role in the control of several cellular processes involved in proliferation, as well as activation of transcription factors, apoptosis and the control of cellular process 2,3 . In addition, the transcription factor c-Myc has been recognized as an important regulator of stem cell biology implicated with GBM malignancy and stemness 4 , as it contributes to proliferation, growth and survival of GBM stem cells 5 . GBM has been also related to the impairment of mitochondrial metabolic capacity, which leads to the alteration in energy production 6,7 and is characterized by an overexpression of Bcl-2 8 . This protein can regulate transition pores permeability of the outer mitochondrial membrane and block pro-apoptotic proteins 9 . Furthermore, it has been identified a novel www.nature.com/scientificreports www.nature.com/scientificreports/ interaction between Bcl-2 and (ADP-ribose) polymerase (PARP) 10 , t...

Amyloid-Beta induces different expression pattern of tissue transglutaminase and its isoforms on Olfactory Ensheathing Cells: modulatory effect of Indicaxanthin

“…In 100 µM Indicaxanthin-treated cells, TG2 staining was slightly higher than controls and it was widely localized in the cytosol. In contrast, in the cells exposed to Aβ(1-42) and Aβ (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35), Indicaxanthin pre-treatment was able to decrease the number of positive cells for TG2, when compared with the controls. Speci cally, in Indicaxanthin treated OECs and subsequently exposed to Aβ(1-42), TG2 was localized into the nucleus.…”

“…Previous our researches demonstrated that TG2 is overexpressed in OECs exposed to Aβ(1-42) and its toxic fragment Aβ (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35) and that the treatment with some Growth Factors (GFs) was able to restore its levels to control values [18]. In particular, TG2, a calcium-dependent protein with transamidanting activity, is involved in AD, inducing the formation of insoluble amyloid aggregates that can alter the properties of several proteins [2].…”

“…In OECs exposed to Aβ (35 − 25), a light modi cation between TG2-L and TG2-S expression levels was observed. The pretreatment with Indicaxanthin was able to counteract the oxidative damage following the exposure of the cells to full native peptide of Aβ and its toxic fragment Aβ (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35), restoring the expression levels of total TG2 to control values. Furthermore, CLSM analysis performed on single cell showed that TG2 in OECs pre-treated with Indicaxanthin alone was localized in the cytosol.…”

“…In the control cells a low staining for TG2 was found and the protein was prevalently localized in the cytosol. A more intense staining for TG2 both in Aβ(1-42) and Aβ (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35) treated OECs was observed widely in the cytosol, when compared with the controls. In particular, the positivity of the cells for TG2 appeared more evident in Aβ(25-35) treated ones, when compared with the controls and Aβ(1-42) treated OECs.…”

“…Since Nestin, marker of neural precursors, is co-expressed in pluripotent stem cells with cyclin D 1 [26], marker of cellular proliferation, the effect of Indicaxanthin pre-treatment on their levels was tested. Furthermore, its effect on OECs exposed to Aβ(1-42), Aβ (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35) and Aβ…”