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. 2021 Sep;29(9):1043-1049.
doi: 10.1016/j.jsps.2021.07.011. Epub 2021 Jul 15.

Neuroprotective effects of Bhilawanol and Anacardic acid during glutamate-induced neurotoxicity

Fadwa Al Mughairbi  1 , Rukhsana Nawaz  1 , Faisal Khan  2 , Amina Hassan  1 , Nailah Mahmood  1 , Heba Tageldeen Ahmed  1 , Alia Alshamali  1 , Sagheer Ahmed  3 , Asma Bashir  4
Affiliations

Neuroprotective effects of Bhilawanol and Anacardic acid during glutamate-induced neurotoxicity

Fadwa Al Mughairbi et al. Saudi Pharm J. 2021 Sep.

Abstract

Bhilawanol (Bh) and anacardic acid (AA) are two lipid-soluble compounds mostly found in the nut of Semecarpus anacardium (SA). This herb has many medicinal properties including enhancing learning and memory, yet its active compounds have not been studied for neuroprotective effects. We investigated the neuroprotective effects of Bh and AA against glutamate induced cell death in the adrenal pheochromocytoma cell line of rats (PC12 cells). Cell viability, toxicity and calcium influx were determined by MTT assay, LDH release assay and Fluo-3 imaging while apoptosis was assayed by caspase-3 and Bcl-2 gene expression. Our results showed that Bh and AA treatments significantly increased cell viability, reduced cell toxicity and calcium influx in PC12 cells in addition to suppressing the reactive oxygen species. Furthermore, AA treatment decreased caspase-3 expression level whereas both Bh and AA enhanced the expression of anti-apoptotic gene Bcl-2 in PC12 cells. Both compounds potently inhibited acetylcholinesterase enzyme (AChE) in a dose and time dependent manner. These findings suggest that the traditional use of SA may be explained on the basis of both Bh and AA showing neuroprotective potential due to their effects on enhancing cell viability, reducing cell toxicity most probably by reducing excessive calcium influx and suppression of ROS as well as by decreasing the expression of proapoptotic caspase 3 gene and increasing the expression of antiapoptotic gene Bcl2. Traditional use in enhancing learning and memory was justified in part by inhibition of AChE.

Keywords: Anacardic acid; Bcl-2; Bhilawanol; Caspase-3; LDH assay; MTT assay; Semecarpus anacardium.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Effects of Bhilawanol and anacardic acid on PC12 cells viability against glutamate toxicity using MTT assay. A) Glutamate toxicity on cell viability. B) Protective effect of Bhilawanol and C) Anacardic acid. Inset showing chemical structure of compound. # p < 0.05 and * p < 0.01 when compared with Glu treated cells and ^ showed significance p < 0.001 between control and glutamate group.
Fig. 2
Fig. 2
Effects of Bhilawanol (Bh) and anacardic acid (AA) on PC12 cell toxicity assessed by LDH assay.*p < 0.01 when compared with Glu treated cells.
Fig. 3
Fig. 3
Effect of AA and Bh on ROS and Calcium influx. A) ROS level in PC12 cells were detected by DCF dye. B) Intracellular level of calcium in PC12 cells was detected by measuring fluorescence of Fluo-3 dye. ^ indicates the p < 0.001 compared with control and * p < 0.001 when compared with Glu treated cells.
Fig. 4
Fig. 4
Bar graph represents % inhibition of compounds on AChE using Ellman’s method. ^ showed significance p < 0.001 compared to control and * indicated the p value < 0.001 as compared to the donepezil group.
Fig. 5
Fig. 5
Expression of apoptotic genes in PC12 cells after treatment. A) Effect of Bh on gene expression. B) Effect of AA on Gene Expression. 1-Control, 2- Glutamate, 3-MK801, 4- Glutamate + Compound, 5- Compound alone.
See this image and copyright information in PMC

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