Searching On The Molecular Mechanism Of The Kai Xin Powder Eliminating Phelgm Sedative Based On The Method Of Serum Pharmacology

Kaixin powder comes from the“emergency prescriptions“, composed of Ginseng,Poria, Radix Polygalae, Acorus tatarinowii Schott, attending “ forget “, The four combinations,nourishing the heart and spleen,eliminating phlegm for resuscitation begins to puzzle, the ancients use it to treating brain diseases, such as the Bureau party and agent stated in the “the heart is not fixed,sorrow and unhappy, forgetfulness,and so on,has a high correlation with Nervous system diseases in modern medicine.Objective:Searching on the molecular mechanism of the Kai xin powder eliminating phelgm sedative based on the method of serum pharmacology. Methods :Firstly,We examined the injury of corticosterone on the expression by gap junction protein of Cx43 by Western-blot;Besides scrape lad dye transfer,we also obsered the change of the gap junction intercellular communication function by inverted fluorescence microscope.Primary cultured astrocytes and neurons used the newborn rats in 24 h,and the neurons will be planted in the 1*1 glass sheet, put the glass plate upside down on the 24 well within the second generation of cultured primary astrocytes.Uptake of upper liquid,measure the content of glutamate and gamma aminobutyric acid by ELISA.CTX TNA2 cell line was used to establish in vitro depression model induced by corticosterone and cell activity was detected with MTT. Then we tested whether Kaixin powder play a neuroprotective role through PI3 K signaling pathway in this cell model with the inhibitors of this pathway and detected by western blotting analysis.Results:According to our data,In vitro model experiment results show that corticosterone after injury, astrocytes activity was inhibited; while adding Kaixin powder containing serum to elevate the survival rate of corticosterone damaged cells; on the basis of the above experiment while giving the PI3 K pathway inhibitor, open heart scattered protective effect was reversed. Western blotting results show that given Kaixin powder containing serum can reverse the corticosterone after injury PI3 K and AKT1 / 2 protein expression decreased. Scratch test, the original generation of astrocytes give different stimulating corticosterone, compared with the control group, fluorescent dye transfer distance becomes shorter with the increase of the concentration of corticosterone, suggesting that the Cx43 protein of gap junction transfer function decreases with the increase of the concentration of corticosterone. And Western blotting results also showed that the with the increase of the concentration of corticosterone and Cx43 protein expression showed an increasing trend,the phosphorylation of Cx43 protein expression showed a downward trend. After co culture model, by giving a happy powder containing serum, ELISA experiment results show that adding Kaixin powder containing different doses of drug serum, compared with the control group, supernatant glutamate release was significantly increased, and gamma aminobutyric acid release no obvious trend.Conclusion:Kaixin powder has a protective effect on corticosterone injury of astrocytes,and the mechanism may be associated with nerve cell protective related PI3 K signal transduction pathway.And Kaixin powder can increase the release of the Glutamate which is the excitatory neurotransmitter,this may be one of mechanisms of antidepressant effect.