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. 2021 May 14:15:651471.
doi: 10.3389/fnins.2021.651471. eCollection 2021.

Ameliorative Role of Cerium Oxide Nanoparticles Against Fipronil Impact on Brain Function, Oxidative Stress, and Apoptotic Cascades in Albino Rats

Norhan Elshony  1 , Atef M K Nassar  2 , Yasser S El-Sayed  3 , Dalia Samak  3 , Ahmed Noreldin  4 , Lamiaa Wasef  1 , Hamida Saleh  3 , Yaser H A Elewa  5   6 , Shereen E Tawfeek  7   8 , Abdullah A Saati  9 , Gaber El-Saber Batiha  1 , Michał Tomczyk  10 , Masakazu Umezawa  11 , Hazem M Shaheen  1
Affiliations

Ameliorative Role of Cerium Oxide Nanoparticles Against Fipronil Impact on Brain Function, Oxidative Stress, and Apoptotic Cascades in Albino Rats

Norhan Elshony et al. Front Neurosci. .

Abstract

Fipronil (FIP) is an N-phenylpyrazole insecticide that is used extensively in public health and agriculture against a wide range of pests. Exposure to FIP is linked to negative health outcomes in humans and animals including promoting neuronal cell injury, which results in apoptosis through the production of reactive oxygen species (ROS). Therefore, the purpose of the current study was to investigate the neuroprotective effects of cerium oxide nanoparticles (CeNPs) on neuronal dysfunction induced by FIP in albino rats. Male rats were randomly classified into four groups: control, FIP (5 mg/kg bwt), CeNPs (35 mg/kg bwt), and FIP + CeNPs (5 (FIP) + 35 (CeNPs) mg/kg bwt), which were treated orally once daily for 28 consecutive days. Brain antioxidant parameters, histopathology, and mRNA expression of genes related to brain function were evaluated. The results revealed oxidative damage to brain tissues in FIP-treated rats indicated by the elevated levels of malondialdehyde (MDA) and nitric oxide (NO) levels and reduced activities of antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPx). On the other hand, the FIP's group that was treated with CeNPs showed decrease in MDA and NO levels and increase in SOD and GPx enzymes activity. Besides, FIP-treated rats showed decreased butyrylcholinesterase (BuChE) activity in comparison to the FIP + CeNPs group. Moreover, FIP caused up-regulation of the expression of neuron-specific enolase (NSE), caspase-3, and glial fibrillary acidic protein (GFAP) but down-regulation of B-cell lymphoma-2 (BCL-2) expression. But the FIP + CeNPs group significantly down-regulated the GFAP, NSE, and caspase-3 and up-regulated the gene expression of BCL-2. Additionally, the FIP-treated group of rats had clear degenerative lesions in brain tissue that was reversed to nearly normal cerebral architecture by the FIP + CeNPs treatment. Immunohistochemical examination of brain tissues of rats-treated with FIP showed abundant ionized calcium-binding adaptor molecule 1 (Iba-1) microglia and caspase-3 and apoptotic cells with nearly negative calbindin and synaptophysin reaction, which were countered by FIP + CeNPs treatment that revealed a critical decrease in caspase-3, Iba-1 reaction with a strong calbindin positive reaction in most of the Purkinje cells and strong synaptophysin reaction in the cerebrum and cerebellum tissues. Based on reported results herein, CeNPs treatment might counteract the neurotoxic effect of FIP pesticide via an antioxidant-mediated mechanism.

Keywords: apoptotic cascades; cerium oxide nanoparticles; fipronil; neurotoxicity; oxidative stress.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
TEM imaging (A) and X-ray EDA pattern of CeNPs nanoparticles (B).
FIGURE 2
FIGURE 2
The effects of FIP, CeNPs, and FIP + CeNPs repetitive doses on lipid peroxidation and antioxidant enzyme activities of the brain tissue of rats. The data was demonstrated as mean ± SEM (n = 7). Means (columns) labeled with different letters (A–D) were significantly different at p ≤ 0.05). CTR, Control; FIP, Fipronil; CeNPs, cerium oxide nanoparticle; MDA, Malondialdehyde; SOD, Superoxide dismutase; GPx, Glutathione peroxidase; NO, Nitric oxide.
FIGURE 3
FIGURE 3
Brain butyrylcholinesterase activity (μmoL/min/mL/mg protein) in control and FIP, CeNPs and FIP + CeNPs-treated groups. Means (charts) of with different label letters (A–D) were varied significantly at p ≤ 0.05. FIP, Fipronil; CeNPs, cerium nanoparticle, and CTR: Control (n = 7).
FIGURE 4
FIGURE 4
Histopathological examination of rat cerebrum. (A) Negative control group. (B) CeNPs group. (C–E) FIP group showing perivascular lymphocytic cuffing (thick arrow in C), neuronal necrosis (thin arrows in D), and neuronophagia (arrows in E). (F) FIP treated with CeNPs. Scale bar = 50 μm.
FIGURE 5
FIGURE 5
Histopathological examination of rat cerebellum. (A) Negative control group with the three layers of the cerebellar cortex from outside to inside; molecular layer (ML), Purkinje cell layer (PL), granule cell layer (GL). (B) CeNPs group. (C) FIP group showing shrunken Purkinje cells (arrows). (D) FIP treated with CeNPs. Scale bar = 50 μm.
FIGURE 6
FIGURE 6
Histochemical staining of rat cerebellum by periodic acid Schiff (PAS). (A) Negative control group with the three layers of the cerebellar cortex from outside to inside; molecular layer (ML), Purkinje cell layer (PL), granule cell layer (GL). (B) CeNPs group. (C) FIP group showing shrunken Purkinje cells (arrows). (D) FIP treated with CeNPs. Normal Purkinje cells (arrowheads), necrotic Purkinje cells (thick arrows), and degenerated Purkinje cells (thin arrow). Scale bar = 50 μm.
FIGURE 7
FIGURE 7
Immunohistochemical staining of rat cerebrum by caspase-3. (A) Negative control group. (B) CeNPs group. (C) FIP group. (D) FIP treated with CeNPs. Notice caspase-3 positive apoptotic neuronal cell nuclei (arrows) with numerous numbers in (C), and few number in (A,B,D). Scale bar = 50 μm.
FIGURE 8
FIGURE 8
Immunohistochemical staining of rat cerebellum by caspase-3. (A) Negative control group. (B) CeNPs group. (C) FIP group. (D) FIP treated with CeNPs. Notice caspase-3 positive apoptotic neuronal cell nuclei (arrows) in all cerebellar cortex layers: molecular layer (ML), Purkinje cell layer (PL), granule cell layer (GL). Numerous number was observed in (C), and few numbers in (A,B,D). Scale bar = 50 μm.
FIGURE 9
FIGURE 9
Immunohistochemical staining of rat cerebrum by ionized calcium-binding adapter molecule 1 (Iba-1). (A) Negative control group. (B) CeNPs group. (C) FIP group. (D) FIP treated with CeNPs. Notice Iba-1 positive microglia (arrows) with numerous numbers in (C), and few numbers in (A,B,D). Scale bar = 50 μm.
FIGURE 10
FIGURE 10
Immunohistochemical staining of rat cerebellum by Iba-1. (A) Negative control group. (B) CeNPs group. (C) FIP group. (D) FIP treated with CeNPs. Notice cerebellar cortex layers: molecular layer (ML), Purkinje cell layer (PL), granule cell layer (GL). Numerous numbers of Iba-1 positive microglia (arrows) was observed in (C), and few number in (A,B,D). Scale bar = 50 μm.
FIGURE 11
FIGURE 11
Immunohistochemical staining of rat cerebellum by calbindin. (A) Negative control group. (B) CeNPs group. (C) FIP group. (D) FIP treated with CeNPs. Notice cerebellar cortex layers: molecular layer (ML), Purkinje cell layer (PL), granule cell layer (GL). Purkinje cells showed calbindin positive reaction (arrows) in (A,B,D). Negative reaction (arrow heads) was observed in all Purkinje cells in (C), and some Purkinje cells in (D). Scale bar = 50 μm.
FIGURE 12
FIGURE 12
Immunohistochemical staining of rat cerebrum by synaptophysin. (A) Negative control group. (B) CeNPs group. (C) FIP group. (D) FIP treated with CeNPs. Scale bar = 50 μm.
FIGURE 13
FIGURE 13
Immunohistochemical staining of rat cerebellum by synaptophysin. (A) Negative control group. (B) CeNPs group. (C) FIP group. (D) FIP treated with CeNPs. Scale bar = 50 μm.
FIGURE 14
FIGURE 14
mRNA relative expression of brain function genes. (A) NSE, neuron-specific enolase. (B) GFAP, glial fibrillary acidic protein in brain tissue of male albino rat exposed to FIP and intoxicated with CeNPs. The expression of transcripts was normalized to GAPDH. The data is demonstrated as mean ± SEM (n = 7). FIP, Fipronil; CeNPs, cerium nanoparticle.
FIGURE 15
FIGURE 15
mRNA relative expression of apoptosis-related genes (A) BCL-2 and (B) caspase-3 in the brain tissues of male albino rats exposed to FIP and intoxicated with CeNPs. The expression of transcripts was normalized to GAPDH. The data is demonstrated as mean ± SEM (n = 7). FIP, Fipronil; CeNPs, cerium nanoparticle.
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