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Ziwuliuzhu Acupuncture Modulates Glu/GABA��?Gln Metabolic Loop Abnormalities in Insomniac Rats

Ao Huang, Dajin Pi, Yiliu Chen, Pin-Hsuan Lee, Yusen Huang, Zuying Hu, Zifeng Zhuang, Peng Qing*, Canghuan Zhao*

Ziwuliuzhu acupuncture is widely considered an effective treatment for insomnia in clinics, but little is known about its possible mechanisms. This study investigated the therapeutic effect of Ziwuliuzhu acupuncture on insomnia and its regulatory mechanism on the Glutamic acid (Glu)/γ-aminobutyric acid (GABA) glutamine (Gln) metabolic loop in a rat model of insomnia. Hematoxylin and eosin staining was used to observe the pathological tissue in the hypothalamus. The levels of the neurotransmitters Glu and GABA in the hypothalamus were determined by high performance liquid chromatography (HPLC) Mass Spectrometry (MS)/MS. Immunohistochemistry (IHC) was used to detect the expression of the GABAA receptor in the hypothalamus. The expression levels of glutamate decarboxylase (GAD65/67) and Glutamine Synthetase (GS) in the hypothalamus were determined by Western blotting. Compared with the model group, the Ziwuliuzhu acupuncture groups showed decreased Glu levels (p<0.001) and GABAA receptor expression (p<0.01), increased GABA levels (p<0.05), and a decreased Glu/GABA ratio. In comparison to the model group, Ziwuliuzhu acupuncture increased the protein expression of GAD65 (p<0.001) and GAD67 (p<0.05) in the hypothalamus and reduced the expression of GS (p<0.01). Glu/GABA-Gln metabolism may be regulated by Ziwuliuzhu acupuncture to produce sedative and hypnotic effects, affecting Glu and GABA synthesis and decomposition, as well as restoring the excitatory/inhibitory balance between Glu and GABA.