Screening And Application Of Monomeric Traditional Chinese Medicine For The Induced Differentiation Of Stem Cells Into Neurocytes

Ischemic cerebrovascular disease is a common and frequently occurring disease which makes harm to human life.Brain injury caused by neuronal cell death or apoptosis is the underlying cause of nerve function insufficiency induced cerebral ischemia. Therefore, the treatment of cerebral ischemia is critical to reduce the injury of nerve function as well as to promote the recovery of neurological function.Because of the poor regeneration ability of neurons in the brain, it is very difficult to recover the neural function after the central nervous system injury. Although there are some neural stem cells in the brain, which can differentiate into neurons,oligodendrocytes and astrocytes,neural stem cells still have its limitations in repairing the nerve injury, because of its limited number and the parts where it can exist in the brain. Bone-Marrow Mesenchymal Stem Cells (BMSCs) are non-hematopoietic stem cells derived from adult cells,which have the ability of self-renewal and differentiation. Studies have shown that BMSCs can differentiate into neural cells including neural stem cell、 neurons and oligodendrocyte glia cells in vitro under specific culture conditions, which can provide favorable evidence for neural mjury repair and regeneration.There are few studies on the differentiation of BMSCs into neurons induced by the traditional Chinese medicine compounds or prescriptions. Therefore, it is important to screen out a kind of traditional Chinese medicine compounds or prescriptions which can induce BMSCs to differentiate into neural cells in finding new drugs to treat cerebral ischemia.In this study, for the first time, we used gene promoter and reporter gene Luc to modify the stable cerebral ischemia drug screening cell model to express the primary screening of gene expression. The primary screening compounds were used to induce BMSCs,real-time quantitative PCR, and immunofluorescence techniques were used to detect expression of five kinds of neural cell markers.In the phase of primary screening,6 compounds of traditional Chinese medicine were screened, and 2 positive compounds were identified to up-regulate gene expression. Bone marrow mesenchymal stem cells were induced by the 2 positive compounds.Results showed that when BMSCs were induced by different concentrations at the same time, relative expression of mRNA of has dose dependent to some extent. When BMSCs were induced by 2 positive compounds of traditional Chinese medicine in concentration of 3μM and 10μM, five kinds of neural cells associated with mRNA relative expression were up-regulated. Furthermore, the mRNA expression amount reached its peak for the first time 72h after BMSC swere induced by the traditional Chinese medicine compounds. BMSCs were induced by 2 compounds for 72h,we have detected the positive rates of neural cell associated with proteins using immunofluorecence.In this study, we have screened 2 compounds of traditional Chinese medicine have the capacity to which can induce BMSCs to differentiate into neural cells,which provides strong evidence on the drug treatment of cerebral ischemia.