The Effects of Hesperidin on BDNF/TrkB Signaling Pathway and Oxidative Stress Parameters in the Cerebral Cortex of the Utero-placental Insufficiency Fetal Rat Model

Abstract

Introduction: Uteroplacental Insufficiency (UPI) produces critical neurodevelopmental problems affecting the Intrauterine Growth Restricted (IUGR) in offspring. This study aimed to investigate the possible neuroprotective roles of Hesperidin (Hes) on the fetal cerebral cortex of the UPI rat model.

Methods: In this experimental study, 40 pregnant Wistar rats (age: ∼40 days, Mean±SD weight: 180±10 g) were randomly divided into 5 groups (n= 8/group). The study groups included control (normal saline, orally), UPI+NS (uterine vessel ligation+normal saline, orally), UPI+HES25, UPI+HES50, and UPI+HES100 (uterine vessel ligation+25, 50 and 100 mg/kg Hes, orally). After being anesthetized by ketamine and xylazine, UPI was induced by permanent bilateral closure of the uterine vessels on Gestation Day (GD) 18. From GD15, the Hes/NS-treated groups received Hes/normal saline until GD21. On GD21, the uterus, placenta, and fetus were dissected out and weighed. The oxidative stress parameters, including Catalase (CAT) activity, Malondialdehyde (MDA), and Total Antioxidant Capacity (TAC) were measured in the fetal cerebral cortex. The expression of Brain-Derived Neurotrophic Factor (BDNF) and Tropomyosin Receptor Kinase B (TrkB) was assessed by RT qPCR methods. The obtained data were analyzed by Analysis of Variance (ANOVA) and Tukey's post hoc test.

Results: The present study findings identified a significant difference in the uterine and fetus weight in Hes-treated mothers (P< 0.05). In the fetus, Hes reduced MDA, and increased CAT activity and TAC (P<0.001 in the UPI+Hes100 group, compared to the UPI+NS group). UPI reduced BDNF and TrkB mRNA expression, compared to the control group (P<0.05). Also, Significant increases in BDNF and TrkB mRNA expression were observed after administrating Hes in the fetal cerebral cortex of the UPI rat model, in a dose-dependent manner (P<0.05).

Conclusion: Hes, as a neuroprotective and antioxidant agent, accelerates BDNF-TrkB signaling pathway and suppresses oxidative stress parameters in the cerebral cortex of the UPI rat model.

Keywords: Brain-derived neurotrophic factor; Hesperidine; Intrauterine growth retardation; Oxidative stress.

Figure 1.

The effect of Hesperidin (H) A: Weight of the uterus; B: The fets; and C: The placenta of the rat (gram). Data are presented as Mean±SD of ten animals. P<0.05 compared to the control group presented by (*), the P<0.05 compared to the UPI+NS group presented by (+); ⁺ P<0.05 and **P<0.001. Abdollahi, H., et al. (2021).

Figure 2.

The oxidative stress parameters in the cerebral cortex The effect of Hes on A: Catalase (CAT, U/ml); B: Malondialdehyde (MDA, μmol/L); and C: Total antioxidant capacity (TAC, mM) levels. Data are presented as Mean±SD. P<0.05 compared to the control group presented by (*), the P<0.05 compared to the UPI+NS group presented by (+), the P<0.05 compared to the UPI+CA25 group presented by (#) and the P<0.05 compared to the UPI+CA50 group presented by ($). ^{+, $} P<0.05; ^{**, ++, ##} P<0.001.

Figure 3.

Gel electrophoresis and effect of hesperidin (Hes) on BDNF and TrkB genes expression in the cerebral cortex of the UPI rat model A: Gel electrophoresis for β2m, BDNF and TrkB genes; B:The BDNF; and C: TrkB genes expression levels were estimated by the quantitative real-time PCR in Hes-treated groups (UPI+Hes25, UPI+Hes50 and UPI+Hes100). Expression data relative to those of the reference gene are presented as Mean±SEM. Statistical significance was tested using the one-way ANOVA. P<0.05 compared to the control group presented by (*), the P<0.05 compared to the UPI+NS group presented by (+), the P<0.05 compared to the UPI+CA100 group presented by (#); ^*,+ P<0.05; ^{**, ++, ##} P< 0.01.