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Pinellia Total Alkaloids Modulate the GABAergic System in Hippocampal Formation on Pilocarpine-Induced Epileptic Rats

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Abstract

Objective

To investigate the neuromodulatory effect of pinellia total alkaloids (PTA) on the gamma-aminobutyric acidergic (GABAergic) system in epileptic rats, and preliminarily evaluate the anti-epileptic effect of PTA.

Methods

Ninety-one male Sprague-Dawley rats were randomized to a control group (n=17) or an epileptic group (n=74) using computer-generated random numbers. Status epilepticus (SE) was induced with pilocarpine in the epileptic group. Epileptic rats that survived SE were randomly divided into 4 groups, namely an epilepsy group (n=13), a topiramate (TPM, 60 mg/kg) group (n=12), a high-dose PTA (800 mg/kg) group (n=12), and a low-dose PTA (400 mg/kg) group (n=10). Treatments were given intragastrically once daily for 14 days. The control group and epilepsy group received normal saline. Spontaneous recurrent seizures (SRSs) were monitored 8-h daily for 7 days after treatment. Then, the hippocampal formation tissues were collected. GABA level was measured using enzyme-linked immunosorbent assay. Protein and mRNA expression levels of glutamate decarboxylase 65 (GAD65), GABA transporter-1 (GAT-1), GABA transaminase (GABA-T), and GABAA receptor (GABAAR) α4, α5, γ2 and δ subunits were measured using Western-blotting analysis and quantitative polymerase chain reaction.

Results

PTA lowered the incidence and frequency of SRS (both doses vs. the TPM group, P>0.05). Compared with the epilepsy group, PTA increased the levels of GABA (both doses P<0.01) and GAD65 (mRNA, 800 mg/kg, P<0.01), and suppressed the levels of GAT-1 (mRNA, 800 mg/kg, P<0.01; 400 mg/kg, P<0.05), GABA-T (mRNA, both doses P<0.01), and GABAAR δ subunit (protein, 800 mg/kg, P<0.05) and γ2 subunit (protein, both doses P<0.01). PTA upregulated the low-expressed mRNA levels of GABAAR α5 subunit (400 mg/kg, P<0.01), δ subunit (800 mg/kg, P<0.05), and γ2 subunit (400 mg/kg, P<0.05).

Conclusions

PTA regulated the GABAergic system through modulating GABA levels and the expression levels of GAD65, GAT-1, GABA-T, and GABAAR α4, α5, γ2 and δ subunits. PTA may exert anti-epileptic effects on the pilocarpine-induced epilepsy model.

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Authors and Affiliations

  1. The First Clinical Medical School, Guangzhou University of Chinese Medicine, Guangzhou, 510406, China

    Chu-xin Deng & Yi Chen

  2. Cerebropathy Center, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510406, China

    Zhi-bing Wu & Zheng-miao Yu

Authors
  1. Chu-xin Deng
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  2. Zhi-bing Wu
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  3. Yi Chen
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  4. Zheng-miao Yu
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Contributions

Deng CX wrote the manuscript. Deng CX and Wu ZB designed the study. Deng CX and Chen Y conducted the experiments. Deng CX, Wu ZB, and Yu ZM conducted the data analysis. All authors read and approved the manuscript.

Corresponding author

Correspondence to Zhi-bing Wu.

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All authors have no conflict of interest to disclose.

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Deng, Cx., Wu, Zb., Chen, Y. et al. Pinellia Total Alkaloids Modulate the GABAergic System in Hippocampal Formation on Pilocarpine-Induced Epileptic Rats. Chin. J. Integr. Med. 26, 138–145 (2020). https://doi.org/10.1007/s11655-019-2944-7

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