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ATAS-EMIT Activates the 5-HT/AC/cAMP Pathway to Ameliorate Insomnia in PCPA Insomniac Rats
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DOI：10.2147/NSS.S536579.png)
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Abstract
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Liangxian Liu,1 Zheng Zuo,2 Zuhong Wang,1 Li Li,1 Yongjiang Fang,1 Chunyan Guo,1 Xuejiao Pan,1 Xiao Xiao1 1Department of Acupuncture, Kunming Municipal Hospital of Traditional Chinese Medicine, Kunming, Yunnan, People’s Republic of China; 2Second Clinical Medical College, Yunnan University of Traditional Chinese Medicine, Kunming, Yunnan, People’s Republic of China Correspondence: Li Li, Department of Acupuncture, Kunming Municipal Hospital of Traditional Chinese Medicine, 25 Dongfeng Dong Lu, Kunming, Yunnan, 663100, People’s Republic of China, Email LiLi200201@sina.com Background: Insomnia may lead to or be co-morbid with mental disorders. However, available treatments have significant side effects. Time-acupoints-space acupuncture, eight methods of intelligent turtle (ATAS-EMIT), as a low-hazard insomnia treatment, has been focused on. Methods: Thirty-six rats were randomly divided into 4 groups: normal (Control), insomnia (Model), ATAS-EMIT treatment (Treatment), and sham acupuncture treatment (Sham). Behavioral experiments were used to assess the treatment of ATAS-EMIT for insomnia. ELISA, Western blot, and immunofluorescence staining were used to detect the expression of 5-HT, AC, cAMP, 5HT1A receptor (5HT1AR), and 5HT2A receptor (5HT2AR) in rat hippocampus. Transcriptome sequencing was used to explore more therapeutic mechanisms. Results: ATAS-EMIT treatment significantly reduced anxiety-like behaviors and activities in PCPA insomniac rats in the open field test. Additionally, ATAS-EMIT significantly shortened the sleep latency period in insomnia-prone rats while prolonging both sleep duration and hanging rest time. These behavioral studies suggest that ATAS-EMIT is a treatment for insomnia. The insomnia-related indicators showed that ATAS-EMIT significantly activated the 5-HT/AC/cAMP pathway, promoted the expression of the 5HT1AR, and inhibited the expression of the 5HT2AR for therapeutic purposes. Subsequent transcriptome sequencing revealed that ATAS-EMIT-treated DEGs were enriched for multiple insomnia-related functions such as phototransduction, stimulus-response, and tryptophan metabolism. Six key genes, Cngb1, Cabp4, Sag, Tyr, Trpm1, and Adipoq, were screened and validated. Conclusion: ATAS-EMIT significantly improved insomnia symptoms in PCPA insomniac rats, and activation of the 5-HT/AC/cAMP pathway was involved. Various mechanisms, such as phototransduction, tryptophan metabolism, and reduction of stimulation, contributed to the therapeutic effects of ATAS-EMIT. Keywords: insomnia, acupuncture, 5-hydroxytryptamine, 5ht1/2a receptor, transcriptome sequencing, mental disorders
Keywords:
insomnia
acupuncture
5-hydroxytryptamine
5ht1/2a receptor
transcriptome sequencing
mental disorders
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