Protective Effect Of Theacrine On HT-22 Cells

Theacrine is a natural purine alkaloid.The structure of it is similar to that of caffeine.Caffeine could stimulate nerves and affects sleep.In contrast to the effects of caffeine,studies have shown that theacrine has sedative and anti-inflammatory effects and could reverse the caffeine-induced decrease in sleep time.In order to explore the effects of theacrine on the nervous system,this experiment took mouse hippocampal neuron cells HT-22 as the research object.First the most suitable cell survival rate detection method was selected to explore the safe dose range of theacrine in cells.Then caffeine was used to induce neuronal hyperexcitation,and the effect of theacrine on neurons of hyperexcitation was investigated.At the cellular level,the effect of theacrine treatment on cell survival and morphology was observed,and changes in intracellular calcium ion content were measured.Transcriptome sequencing is performed at the molecular level to analyze which genes,biological processes,and pathways were regulated by theacrine.The CTG method was used to test the cell survival rate through experiments.It is determined that the safe concentration of theacrine is 0.2mM to 1mM,and the concentration of caffeine-induced over-excitation of HT-22 cells is 50mM for half an hour.It was found that although theacrine could not increase the cell survival rate after caffeine intervention,it could reduce the nerve cell aberration induced by caffeine.The caffeine intervention significantly increased the intracellular calcium ion content to 248.8nM,which is almost 6 times of it in control samples.On the basis of this,the intervention of 0.2mM theacrine could reduce the elevated calcium ion content and make it close to the control samples.Thereby,theacrine could reduce the hyperexcitation of the cells and protect the nerve cells.Through the analysis of transcriptome data,it was found that theacrine could regulate some biological processes and gene expression in cells,it is speculated that the reduction of calcium content by theacrine may be achieved by mediating GPCR/Ca²⁺/PKC pathway.