Screening Of Active Ingredients For The Treatment Of Insomnia With Shuangxia Decoction Targeting The Serotonin 1A Receptor

Insomnia is a common disease and frequently encountered in our life. It usually has a longer duration and is difficult to cure. Severe cases may also induce or aggravate other diseases such as palpitation, chest obstruction, dizziness, stroke, etc. In western medicine, hypnotic drugs such as benzodiazepine were used most often for curing insomnia. Despite the advantages of speedness, but they also had side effects like tolerance and drug addiction and so on. Chinese medicine for insomnia has a long history and handed down a lot of classic prescriptions for curing insomnia, one of which is the Shuangxia soup. In recent years, many reports about the prescription were published in clinical treatment and showed a good therapeutic effect. However, further exploitation of Shuangxia soup is serious prevented since the material basis of Shuangxia soup for hypnosis is not clear enough.With the progress of molecular biology in pharmacology, more and more researchers prefer to researching chemical materials on the basis of receptors. The reason why it attracts much more pro-gaze is that many advantages such as good specificity and rapidness and so on can be acquired. For insomnia drug research,5-HTIA receptor has been shown to correlate with sleep. Exciting 5-HTIA receptor can produce certain sedative and hypnotic effects. Therefore, the following experiments were done.(1) According to the chemical properties (such as acid and alkali, polarity, etc.) of composition, chemical composition groups of total flavonoids, total alkaloid, total saponins were separated from Shuangxia soup. Chemical composition groups separated from Shuangxia Soup were composed. The 6 samples were got. In order to evaluate the exciting activity in 5-HTIA receptor of inspected samples, fluorescence signal generated by combination of intracellular calcium ion and fluorescent agent was detected using Calcium current detection device (Flex Station II) after HEK293/5-HTRIA cells were stimulated by samples. Results:Flavonoids generated a positive result in filter components. Compared with negative control group, the fluorescent signal of Flavonoids enhanced obviously. It suggests that Flavonoids can excite 5-IITIA receptor.(2)Systematic separation was carried out on the Shuangxia soup according to the polarity:The Chinese material medicine (pinellia and selfheal were mixed as the rate of 1:1) were extracted by 70% alcohol, and then concentrated into extractum. Column chromatography was used for separation and thin layer chromatography for identification. Then the same or similar fractions were merged. The isolated fractions were completed the activity screening of agonist for 5-HTIA receptor. In order to evaluate the exciting activity in 5-HTIA receptor of inspected samples, fluorescence signal generated by combination of intracellular calcium ion and fluorescent agent was detected using calcium current detection device (Flex Station II) after HEK293/5-HTRIA cells were stimulated by samples. Results:Compared with negative control group, the fluorescent signal of fractions that has larger polarity enhanced obviously and may be 5-HTIA receptor agonist.(3)As total flavonoids are a hit in the primary screen, extraction method of total flavonoids is optimized by response surface method. The actual withdrawal rate is up to 1.84 mg/g.