Ziwuliuzhu Acupuncture Modulates Clock mRNA, Bmal1 mRNA and Melatonin in Insomnia Rats

Abstract

Background: In clinics, Ziwuliuzhu acupuncture is widely considered an effective method of treating insomnia; however, there is currently limited information available regarding its possible mechanisms. Although the method of Ziwuliuzhu acupuncture possesses a unique rhythmic pattern. Objectives: In this study, we have creatively combined the traditional Chinese medicine of Ziwuliuzhu with a modern biological rhythm to investigate the internal mechanism of insomnia. Methods: Pathological tissue from the hypothalamus was analyzed using hematoxylineosin staining. The level of TNF (tumor necrosis factor)-α in the SCN (suprachiasmatic nucleus) area of the hypothalamus was detected in situ using the TUNEL fluorescence staining assay. The concentration of hypothalamic melatonin was detected using the enzyme-linked immunosorbent assay (ELISA). The mRNA expression of Clock and Bmal1 was measured using RT-qPCR. Results: In the Ziwuliuzhu acupuncture groups, the structural damage in the hypothalamic neurons was alleviated compared to the model group and the expression of inflammatory factors was reduced. The mRNA expression levels of Clock and Bmal1 were significantly increased (p < 0.05). The concentration of melatonin was significantly increased (p < 0.001). Although there were no significant differences between the treatment groups (diazepam group, Nazi group, Najia group, and routine group) (p > 0.05). Conclusion: Ziwuliuzhu acupuncture alleviated neuronal damage and modulated the inflammatory reaction in the hypothalamus of rats with insomnia. Moreover, Ziwuliuzhu acupuncture increased the expression levels of Clock and Bmal1 mRNA, and MT content. This study has potentially highlighted one of the mechanisms through which Ziwuliuzhu acupuncture can be used to treat insomnia.

Acupuncture in circadian rhythm sleep–wake disorders and its potential neurochemical mechanisms

“…Acupuncture regulates circadian rhythms by regulating the expression of SCN-associated circadian clock genes and proteins. In addition to reducing sleep latency and lengthening sleep time, a study found that acupuncture for insomniac male Sprague Dawley (SD) rats can also reduce the expression level of tumor necrosis factor-α (TNF-α), thereby alleviating inflammatory responses and decreasing structural damage in the hypothalamus ( Wei et al, 2017 ; Huang et al, 2023 ). Additionally, it can increase the expression levels of Per1 and Per2 mRNA, as well as Clock and Bmal1 mRNA ( Guo et al, 2017 ).…”

“…Through raising MT levels and lowering TNF-α expression in hypothalamic SCN cells, acupuncture can alleviate the pathological products and inflammatory responses of chronic insomnia ( Huang et al, 2023 ). Acupuncture has a particularly good safety profile and has been shown in a trial to alleviate insomnia in pregnant women by elevating melatonin levels without any significant adverse events ( Foroughinia et al, 2020 ).…”

“…For example, MT treatment can lower the methylation level of the circadian clock gene Per1 to promote its rhythmic expression (Tsukamoto-Yamauchi et al, 2015), and can also increase Bmal1 expression via various signaling pathways (Sanchez et al, 2018). Through raising MT levels and lowering TNF-α expression in hypothalamic SCN cells, acupuncture can alleviate the pathological products and inflammatory responses of chronic insomnia (Huang et al, 2023). Acupuncture has a particularly good safety profile and has been shown in a trial to alleviate insomnia in pregnant women by elevating melatonin levels without any significant adverse events (Foroughinia et al, 2020).…”