Study On The Effective Ingredients In Gastrodia Targeting MT₁ To Exert Sedative And Tranquilizing Effects

Sleep has been an essential stage in the daily life of modern humans.About one-third of our body is in a state of sleep in our life.The sleep stage has the function of restoring physical strength,eliminating fatigue,promoting physical development and growth,and enhancing learning and memory.Therefore,sleep is of great significance to us.With the rapid development of science and technology.With the rapid development of science and technology,Affected by one’s own endogenous pressure and exogenous environmental factors,insomnia that is manifested as difficulty in falling asleep,decreased sleep quality,and easy awakening during sleep has become more and more common in society.Short-term insomnia can cause fatigue,exertion,and lack of concentration,while long-term insomnia can cause depression,decreased immunity,and metabolic diseases.Therefore,the problem of insomnia cannot be ignored,and it has received more and more attention.Benzodiazepines are commonly used for the treatment of insomnia,but they have many side effects and great harm.In China,traditional Chinese medicine has been used to treat diseases since ancient times.Gastrodia has been proven to calm and soothe the nerves,relieve wind and relieve spasms,and treat dizziness,epilepsy,insomnia and other diseases.However,the diverse and complex compounds in Gastrodia and the numerous physiological effects hinder the development of new drugs.Therefore,The research direction of this article selects the active ingredients in Gastrodia,including gastrodin,Parishin A and Parishin B,as well as the sleep system-related receptors that our laboratory has constructed,such as dopamine receptor D₂,D₃,orexin receptor OX₂,melatonin receptor MT₁,MT₂high-expressing cell line,as a platform to screen potential targets that may play a sedative and soothing mechanism.According to the experimental results of this study,it is shown that gastrodin and Parishin A can up-regulate the p-ERK protein level through the MT₁receptor and is dependent on the compound concentration.Then the thermal stability experiment,protease cleavage experiment and the influence of the receptor probe FRET baseline further verified the binding ability of Parishin A and the MT₁receptor.Then the thermal stability experiment,protease cleavage experiment and the influence of the receptor probe FRET baseline further verified the binding ability of Parishin A and the MT₁receptor.Finally,after point mutations in the MT₁receptor binding pocket,Parishin A cannot activate ERK1/2 protein through it,indicating that Parishin A specifically binds to MT₁receptors.Gastrodin,Parishin A and Parishin B were administered to mice by nasal injection to induce sleep in mice.It was found that gastrodin and Parishin A can effectively prolong the sleep time induced by pentobarbital sodium,shorten the sleep latency of mice,and reduce the spontaneous activity of mice.At the same time,the detection of DA and5-HT by ELISA shows that gastrodin and Parishin A can significantly increase the content of DA and 5-HT in the brain of mice.Through q-PCR experiments,it was found that gastrodin and Parishin A can reduce the expression of inflammatory factors in the liver and kidney,but can up-regulate the expression of fos gene,suggesting that these two compounds can activate neurons in mice and reduce inflammation factors indirectly promote sleep and at the same time play a role in neuroprotection.The routine blood biochemical test found that the three compounds had no effect on the blood biochemical indicators of mice.