Chaihu-Longgu-Muli decoction relieves epileptic symptoms by improving autophagy in hippocampal neurons

J Ethnopharmacol. 2020 Sep 15:259:112990. doi: 10.1016/j.jep.2020.112990. Epub 2020 May 19.

Abstract

Ethnopharmacological relevance: Chaihu-Longgu-Muli decoction (CLMD) is a well-known ancient formula in traditional Chinese medicine (TCM) to relieve disorder, clear away heat, tranquilize the mind and allay excitement. It has been used for the therapy of neuropsychiatric disorders such as epilepsy, dementia, insomnia, anxiety, and depression for several centuries in China.

Aim of the study: This paper is based on the assumption that the mechanism by which CLMD relieves epileptic symptoms in rats is associated with improving autophagy. Several experimental methods are designed to testify the hypothesis.

Materials and methods: The lithium-pilocarpine-induced epilepsy model was established in rats. The seizure frequency was recorded. Morphology and number of autophagosomes in hippocampal dentate gyrus was detected with a transmission electron microscope (TEM). Expression of Beclin-1, microtubule-associated proteins 1A/1B light chain 3 (LC3), and mammalian target of rapamycin (mTOR) in dentate gyrus was measured by immunofluorescence assay, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western-blotting.

Results: CLMD could significantly relieve the seizure frequency and improve autophagy in hippocampal dentate gyrus. Meanwhile, the level of Beclin-1 and LC3B decreased significantly, while mTOR increased remarkably after medical intervention.

Conclusions: CLMD could improve autophagy in hippocampal dentate gyrus due to epilepsy, especially at high dose. The mechanism may be related to upregulated expression of mTOR and downregulated expression of Beclin-1 and LC3B.

Keywords: Autophagy; Chaihu-Longgu-Muli decoction (CLMD); Epilepsy; Hippocampus; Status epilepticus.

MeSH terms

  • Animals
  • Anticonvulsants / pharmacology*
  • Autophagosomes / drug effects
  • Autophagosomes / metabolism
  • Autophagosomes / ultrastructure
  • Autophagy / drug effects*
  • Autophagy-Related Proteins / genetics
  • Autophagy-Related Proteins / metabolism
  • Behavior, Animal / drug effects
  • Disease Models, Animal
  • Drugs, Chinese Herbal / pharmacology*
  • Epilepsy / chemically induced
  • Epilepsy / drug therapy*
  • Epilepsy / metabolism
  • Epilepsy / pathology
  • Hippocampus / drug effects*
  • Hippocampus / metabolism
  • Hippocampus / physiopathology
  • Hippocampus / ultrastructure
  • Lithium Chloride
  • Male
  • Neurons / drug effects*
  • Neurons / metabolism
  • Neurons / pathology
  • Pilocarpine
  • Rats, Sprague-Dawley
  • Signal Transduction

Substances

  • Anticonvulsants
  • Autophagy-Related Proteins
  • Drugs, Chinese Herbal
  • chaihu-longgu-muli decoction
  • Pilocarpine
  • Lithium Chloride