Effect of Tuina on Hypothalamic Orexin-A Expression and Sleep Behavior in a Rat Model of Primary Insomnia

Insomnia is a common sleep disorder that leads to impaired daytime function and increased risk of comorbidities, significantly impacting patients' quality of life. Current treatments, primarily cognitive behavioral therapy and pharmacotherapy, are limited by issues such as drug dependence, tolerance, and withdrawal rebound. Tuina therapy has been shown to regulate the nervous, endocrine, and immune systems, while the orexin system is a key regulator of the sleep-wake cycle. This has prompted the exploration of whether Tuina alleviates insomnia by modulating the hypothalamic orexin system. This protocol describes the methods of Tuina intervention in a rat model of primary insomnia induced by the modified multiple platform water environment method. We randomized 64 Wistar rats into four groups: control, model, Tuina, and orexin antagonist. Behavioral assessments (open field test, pentobarbital-induced sleep test) were conducted, and the expression of Orexin-A in the hypothalamus was detected via real-time quantitative PCR and immunohistochemistry. The protocol aims to evaluate the efficacy of Tuina and investigate its potential mechanism related to the orexin system, providing a reference for the application and mechanistic study of Tuina in sleep disorders.