The Modulation of NMDA and AMPA/Kainate Receptors by Tocotrienol-Rich Fraction and Α-Tocopherol in Glutamate-Induced Injury of Primary Astrocytes.

Astrocytes are known as structural and supporting cells in the central nervous system (CNS). Glutamate, as a main excitatory amino acid neurotransmitter in the mammalian central nervous system, can be excitotoxic, playing a key role in many chronic neurodegenerative diseases. The aim of the current study was to elucidate the potential of vitamin E in protecting glutamate-injured primary astrocytes. Hence, primary astrocytes were isolated from mixed glial cells of C57BL/6 mice by applying the EasySep® Mouse CD11b Positive Selection Kit, cultured in Dulbecco's modified Eagle medium (DMEM) and supplemented with special nutrients. The IC20 and IC50 values of glutamate, as well as the cell viability of primary astrocytes, were assessed with 100 ng/mL, 200 ng/mL, and 300 ng/mL of tocotrienol-rich fraction (TRF) and alpha-tocopherol (α-TCP), as determined by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The mitochondrial membrane potential (MMP) detected in primary astrocytes was assessed with the same concentrations of TRF and α-TCP. The expression levels of the ionotropic glutamate receptor genes (Gria2, Grin2A, GRIK1) were independently determined using RT-PCR. The purification rate of astrocytes was measured by a flow-cytometer as circa 79.4%. The IC20 and IC50 values of glutamate were determined as 10 mM and 100 mM, respectively. Exposure to 100 mM of glutamate in primary astrocytes caused the inhibition of cell viability of approximately 64.75% and 61.10% in pre- and post-study, respectively (p < 0.05). Both TRF and α-TCP (at the lowest and highest concentrations, respectively) were able to increase the MMP to 88.46% and 93.31% pre-treatment, and 78.43% and 81.22% post-treatment, respectively. Additionally, the findings showed a similar pattern for the expression level of the ionotropic glutamate receptor genes. Increased extracellular calcium concentrations were also observed, indicating that the presence of vitamin E altered the polarization of astrocytes. In conclusion, α-TCP showed better recovery and prophylactic effects as compared to TRF in the pre-treatment of glutamate-injured primary astrocytes.

Lethal and Sublethal Effects of Essential Oils From Artemisia khorassanica and Vitex pseudo-negundo Against Plodia interpunctella (Lepidoptera: Pyralidae).

Plodia interpunctella (Hübner, 1813) is a polyphagous and key pest of different stored products worldwide. The lethal and sublethal effects of essential oils of Artemisia khorassanica Podl. and Vitex pseudo-negundo (Hausskn) were studied on P. interpunctella The chemical constituents of the essential oils were also assessed using gas chromatography-mass spectrometry. Assays showed that the fumigant toxicity of A. khorassanica (LC50: 9.60 µl/liter air) was higher than V. pseudo-negundo (LC50: 23.05 µl/liter air). Moreover, the speed of mortality caused by A. khorassanica oil (LT50: 2.07 h) was higher than V. pseudo-negundo (LT50: 3.11 h). To assess the sublethal effects of the essential oils, adult moths were exposed to the LC30 of each essential oil, and life table parameters and energy contents of the surviving P. interpunctella were studied. Exposure to sublethal concentration of A. khorassanica negatively affected the life table of P. interpunctella, and also the protein, lipid, and glycogen contents of the larvae that came from treated adults. Vitex pseudo-negundo also affected lipid, protein, and glycogen contents of P. interpunctella The intrinsic rate of increase (rm), finite rate of increase ([Formula: see text]), and doubling time (DT) were not significantly different between control and V. pseudo-negundo treatment. According with these results, both tested essential oils, especially one extracted from A. khorassanica, have potential applications for the integrated management of P. interpunctella.

A Validated HPTLC-Densitometric Method for Assay of Aucubin in Vitexagnus-castusL.

Vitexagnus-castusL. is a medicinal plant which is used in several dosage forms for women hormonal disorders and standardized according to the iridoids or flavonoids content. Aucubin, an iridoid glycoside, considered as a marker in some formulations. In this research, a thin layer chromatographic method with densitometric detection has been developed for quantitative determination of aucubin in chaste tree fruits. Chromatographic separation was performed using silica gel high performance thin-layer chromatography (HPTLC) plates with ethyl acetate-methanol-water 77 : 15 : 8 as mobile phase. Chromatograms were visualized using p-dimethylaminobenzaldehyde as reagent. Aucubin RF-value was about 0.5 and spots were scanned at 580 nm through a mercury lamp. By using this method, the amount of aucubin was found 43.5 mg/100 g of dried plant fruits. The method was validated for selectivity, linearity (r(2) = 0.997, 20-100 µg/mL), precision (intra-day < 4.9, inter-day < 7.2) and accuracy measured via determination of recovery (95-98%). The limit of detection and limit of quantization were found 6.6 and 20 µg/mL, respectively. This methodology was found to be precise with respect to the validation parameters. It is simple and convenient and could be applicable to the routine determination of aucubin in different Vitexagnus-castusL. samples.