High Throughput Screening Of Agonist For M1 Receptor From Traditional Chinese Medicine

Alzheimer's disease(AD) is an ageing disease of high incidence in modern society. The symptom are damage of cholinergic system and formation of amyloid spots in the brain. Muscarinic M1 receptor has close relation to AD since the activation of M1 receptor can inhibit the formation of amyloid spot and improve the function of cholinergic system. However,the agonists for M1 receptor in AD therapy have low selectivity which cause a lot of side effects. So screening new selective agonist for M1 receptor is an important field in AD therapy. Animal screening is the traditional screening method with hard work and low efficiency. Recently, with the development of Biomolecular, pharmacology, automatic et al, high throughput screening(HTS) technology appeared with high efficiency using molecular or cell screening targets in minimal form. This paper established a screening method for high throughput screening of agonist for M1 receptor and screened 400 extracts of Traditional Chinese medicine. This screening method couples M1 receptor to reporter gene, it mean, M1 receptor gene and reporter gene are cotransfected HEK293 cell line to establish a stable cell line for screening. When a compound binds to M1 receptor, it will activate M1 receptor and the signal pathway to change the expression of the second messagers in the cell. Responsive elements respond to this change and cause the expression of reporter gene. The bioactivity of the compound can be evaluated after the expression of the reporter gene was measured. Using this cell line, 400 extracts of Traditional Chinese medicine(TCM) were screened and three TCM were found to activate M1 receptor. One of the TCM was separated to find the compound of the bioactivity. In a word, this screening model can be succeeded in screening agonist for M1 receptor in high throughput screening and identifying the bioactivity components of Traditional Chinese medicine.