CXCR4 antagonist AMD3100 attenuates epileptic activity by enhancing inhibitory neurotransmission in rats
10.3760/cma.j.cn115354-20210815-00513
- VernacularTitle:CXCR4拮抗剂AMD3100通过增强抑制性神经传递减弱大鼠痫性活动
- Author:
Yusong ZHANG
1
;
Zhiguo CHEN
;
Zishan YANG
;
Lei LI
Author Information
1. 郑州大学附属肿瘤医院（河南省肿瘤医院）病理科，河南省肿瘤病理和人工智能诊断医学重点实验室，郑州 450000
- Keywords:
Epilepsy;
Gama-aminobutyric acid;
C-X-C chemokine receptor type 4;
AMD3100
- From:
Chinese Journal of Neuromedicine
2022;21(1):6-12
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigative the molecular mechanism of C-X-C chemokine receptor type 4 (CXCR4) antagonist AMD3100 in epileptic seizure.Methods:(1) Animal experiment: 36 adult male SD rats were randomly divided into control group (Con, n=12), epilepsy group (Epi, n=12), Epi+AMD3100 group ( n=12). Experimental epilepsy rat models in the Epi group were induced by intraperitoneal injection of pentrazole (PTZ, 40 mg/kg); rats in the Epi+AMD3100 group were given intraperitoneal injection of PTZ (40 mg/kg) 20 min after lateral intracerebroventricular injection of 5 μL (5 mg/mL) AMD3100; rats in the Con group were given intraperitoneal injection of normal saline. Racine grading was used to evaluate the levels of epileptic seizure and the latency of epileptic seizure was recorded in rats from each group. EEG was used to record the abnormal discharges of brain neurons in rats from each group. The content of γ -aminobutyric acid (GABA) in the hippocampus was detected by ELISA kit; γ -aminobutyric acid A receptor α1 subunit ( GABAAR α1) mRNA levels of hippocampal neurons in each group were detected by real-time fluorescent quantitative PCR (qRT-PCR). (2) Cell experiment: the hippocampal neurons from 1-d-old SD rats were primarily cultured; 7 d after cultivation, these cells were divided into control group, epilepsy group and AMD3100 group; the cellular epileptic models in the epileptic group were induced by magnesium-free external fluid; neurons in the AMD3100 group were cultured in magnesium-free external solution containing 10 nmol/L AMD3100 for 3 h, and then changed to Neurobasal medium for further culture; cells in the control group were cultured with Neurobasal medium. Whole cell patch clamp technique was used to detect the spontaneous inhibitory postsynaptic currents (sIPSCs) after AMD3100 (10 nmol/L) perfusion. Results:(1) Animal experiment: the seizure latency in Epi+AMD3100 group was significantly shorter than that in Epi group ([663.30±74.84] s vs. [164.40±17.20] s, t=6.490, P<0.001). The frequency of seizures>grading 4 in the Epi+AMD3100 group was significantly decreased as compared with that in the Epi group (3.75±0.39 vs. 9.00±0.73, t=4.680, P<0.001). ELISA results showed that GABA content in the 3 groups was significantly different ( F=17.850, P<0.001): that in the Epi group was significantly lower than that in the Con group, and that in the Epi+AMD3100 group was significantly higher than that in Epi group ( P<0.05). The qRT-PCR results showed that GABAAR α1 mRNA content among the 3 groups was significantly different ( F=14.400, P<0.001): that in the Epi group was significantly lower than that in the Con group, and that in the Epi+AMD3100 group was significantly higher than that in the Epi group ( P<0.05). EEG results showed that the discharge frequency of rats in the Epi+AMD3100 group was lower than that in Epi group; there was no significant difference in EEG power among the 3 groups ( F=3.220, P<0.001), but the EEG power in the Epi+AMD3100 group was lower than that in Epi group and control group. (2) Cell experiment: patch clamp technique showed that the average frequency and amplitude of sIPSCs in the 3 groups were statistically significant ( F=13.670, P<0.001; F=10.920, P<0.001). As compared with those in the control group and epilepsy group, the average frequency and amplitude of sIPSCs in AMD3100 group were significantly increased ( P<0.05). Conclusion:CXCR4 antagonist AMD3100 can reduce seizure frequency by enhancing inhibitory neurotransmission.
