ABSTRACT
Trigonelline-loaded water-soluble chitosan nanoparticles (Trigo-WSCS NPs) were prepared for the treatment of glioblastoma (targeting C6 glioma cells) and also evaluated its biocompatibility with rat adrenal pheochromocytoma cells (PC12 cells). WSCS-Trigo NPs characteristics were determined using UV-Visible spectrophotometer, FTIR, XRD, TEM, DLS, and Zeta potential. Trigo-WSCS NPs were noted to have a spherical shape, with an average size of 356 nm. Trigo-WSCS NPs zeta potential was 30.9 mv, which expresses its good stability. The WSCS-Trigo NPs considerably inhibited the growth of rat C6 glioma cells and exhibited an IC50 concentration of 34 µg/mL. Further, Trigo-WSCS NPs were biocompatible with PC12 cells in terms of enhancing neurite growth and differentiation. In conclusion, Trigo-WSCS NPs could act as an antitumor drug for the treatment of glioblastoma as suggested by the in vitro studies.
Subject(s)
Alkaloids/chemistry , Alkaloids/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Chitosan/chemistry , Drug Carriers/chemistry , Nanoparticles/chemistry , Adrenal Gland Neoplasms/metabolism , Alkaloids/administration & dosage , Animals , Antineoplastic Agents/administration & dosage , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Chitosan/administration & dosage , Drug Carriers/administration & dosage , Drug Carriers/chemical synthesis , Drug Liberation , Flow Cytometry , Glioblastoma/drug therapy , Glioblastoma/metabolism , Glioblastoma/pathology , Glioma/drug therapy , Glioma/metabolism , Glioma/pathology , Materials Testing , Microscopy, Fluorescence , Nanoparticles/administration & dosage , Nanoparticles/analysis , Particle Size , Pheochromocytoma/metabolism , RatsABSTRACT
Asiaticoside (AS), a triterpenoid saponin isolated from the Indian medicinal herb Centella asiatica is known to exert a neuroprotective effect by attenuating the neurobehavioral, neurochemical and pathological changes in animal models. However, its potential neuroprotection in rotenone-induced hemiparkinsonism which implicates phospholipid-mediated neurotransmission remains unclear. Therefore, we have investigated the neuroprotective effects of AS in rat model of ROT-infused hemiparkinsonism with respect to phosphoinositides-assisted cytodynamics and synaptic function. Adult male Sprague-Dawley rats (250-300g) were distributed randomly into 6 groups, with 6 rats in each group: Sham control, Vehicle control (DMSO-0.1%), ROT-infused group (6µg/µl/kg), AS-treated group (50mg/kg/day), Drug (AS) control and Levodopa (l-DOPA)-treated group (6mg/kg/day). At the end of the experimental period, the rats were sacrificed after performing behavioral analyses and the striatum regions were dissected out. Phosphoinositides (PI) are involved in intrinsic membrane signals that regulate intracellular membrane trafficking vesicle and endocytosis. We have assessed mRNA and protein expressions of genes involved in PI-mediated signaling and also in synaptic function (PI3K, PDK 1, PEBP, Stx 1A and TH) in addition to the levels of neurotransmitters and the enzymatic antioxidant profile. AS caused an improved working memory and motor co-ordination in the ROT group. It alters the levels of neurotransmitters (p<0.01), the expression of mRNA and protein assessed which were significantly affected (P<0.001) by rotenone, thus exhibiting its intervention in the progression of neurodegeneration. We demonstrate that AS can mediate distinct function in PI-assisted vesicle endocytosis, cytoprotective signaling and in the synaptic function thereby mitigating the ROT-infused hemiparkinsonism, however, its specific regulatory role remains to be unraveled.
Subject(s)
Disease Models, Animal , Oxidative Stress/drug effects , Parkinsonian Disorders/prevention & control , Phosphatidylinositols/metabolism , Rotenone/pharmacology , Synapses/drug effects , Triterpenes/pharmacology , Animals , Behavior, Animal/drug effects , Blotting, Western , Chromatography, High Pressure Liquid , Male , Maze Learning , Parkinsonian Disorders/chemically induced , Parkinsonian Disorders/metabolism , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Rotarod Performance Test , Synapses/metabolismABSTRACT
Parkinson's disease (PD), a progressive neurodegeneration, is characterized by loss of dopaminergic neurons in the substantia nigra (SN) and loss of motor co-ordination. Impaired metabolism of major lipids such as phospholipids which play regulatory roles in cellular functions and signaling has been implicated in the pathology of PD. We aim to investigate the striatal phospholipids (PLs) in hemiparkinsonism infused by rotenone in rats. As there are no cost-effective modes of PL, we have utilized dye-lipid complex technique for the first time in PD models for screening and also for semi-quantifying (individually) the levels of the deregulated PL in brain samples. Rats were divided into 2 groups: i. control and ii. ROT-infused which received intracranial injection of Rotenone (6 µg/µl; flow rate 0.2 µl/min). At the end of experimental period of 14 days, the striatum was dissected out for the analyses of PLs. Dye-based detection of PL and two-dimensional thin-layer chromatographic analyses of PL were performed. Detection of dye-PL complex was possible for phosphatidyl choline (PC), phosphatidyl inositol (PI), and spingomyelin (SM) (but not for phosphatidyl ethanolamine-PE) using dyes viz victoria blue B, toluidine blue and ammonium ferrothiocyanate, respectively. Two-dimensional analyses of phospholipids confirmed the dye-PL complex and depicted significant reduction (p < 0.05) on semi-quantitative assessment, in the striatum of control and hemiparkinsonic rats. We suggest a low level of PLs esp of PI in striatum of rats using a simple dye-detection that was validated by HR-LCMS. The finding implies that a critical role is being played by these PLs (PC, PI and SM) mainly PI (p < 0.001), in rotenone infused hemiparkinsonism, thus deserving wider but simpler investigations to detect and identify their role in parkinsonism.
