Ping Yang1, You Qin2, Yong Zhu3, Feng Li4, Shuai-Shuai Xia5, Bin Zhou3, Qin Wang3, Jun Lu3, Liang Li6, Hui-Yong Huang7. 1. Department of Psychiatry, Hunan Brain Hospital, Clinical Medical School of Hunan University of Chinese Medicine, NO.427, Middle Furong Road, Changsha, Hunan Province, 410007, China; Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, NO.300, Xueshi Road, Changsha, Hunan Province, 410208, China. 2. Institute of Chinese Materia Medica, Hunan Academy of Chinese Medicine, NO. 8, Yuehua Road, Changsha, Hunan Province, 410013, China. 3. Department of Psychiatry, Hunan Brain Hospital, Clinical Medical School of Hunan University of Chinese Medicine, NO.427, Middle Furong Road, Changsha, Hunan Province, 410007, China. 4. Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, NO.300, Xueshi Road, Changsha, Hunan Province, 410208, China; School of Dentistry, University of California Los Angeles, 405 Hilgard Ave, Los Angeles, CA, 90095, USA. 5. Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, NO.300, Xueshi Road, Changsha, Hunan Province, 410208, China. 6. Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, NO.300, Xueshi Road, Changsha, Hunan Province, 410208, China. Electronic address: liliang@hnucm.edu.cn. 7. Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, NO.300, Xueshi Road, Changsha, Hunan Province, 410208, China. Electronic address: huanghy@hnucm.edu.cn.
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.
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.