| Background Mesenchymal stem cells(MSCs),as ideal seed cells for cell therapy,have been widely used to repair various tissue and organ injuries caused by clinical injury,aging and other diseases after long-term in vitro and animal disease model therapy.Tissue repair mainly includes bone reconstruction and regeneration,neurodegenerative diseases and injuries,myocardial injury,liver cirrhosis,diabetes mellitus,lung diseases and so on.Mesenchymal stem cells come from a wide range of sources,such as spinal cord mesenchymal stem cells,umbilical cord blood mesenchymal stem cells,placental mesenchymal stem cells,adipose mesenchymal stem cells and so on.How to differentiate mesenchymal stem cells into neuron-like cells and provide a safe induction method for clinical use is a research hotspot in the field of spinal cord injury(SCI).Mongolian medicine is a summary of Mongolian people's experience in fighting diseases,and gradually formed by absorbing the experience of Tibetan medicine and traditional Chinese medicine.It is a cultural treasure of our country.Among them,Zhenbao pills,a Mongolian medicine,is widely used in clinical practice,which plays a neuroprotective role after nerve injury,and plays a certain role in the repair of central nervous system and peripheral nervous system.The results showed that Zhenbao Pill had certain effects on the proliferation of neuron-like cells and had protective effects on the nerves of rats with acute spinal cord injury.Therefore,this article will discuss the effect of Mongolian medicine Zhenbao Pills on the directional differentiation of human adipose mesenchymal stem cells into neurons.ObjectiveAdipose-derived mesenchymal stem cells(ADMSCs)were extracted from adipose tissue collected during operation by serum culture containing Mongolian medicine Zhenbao pills.To explore the effect of Zhenbao Pills on directional differentiation of mesenchymal stem cells into neural stem cells,and to find a new,reliable and safe inducer.To create convenience for transplantation of mesenchymal stem cells to treat spinal cord injury,and provide theoretical basis for Zhenbao Pills to differentiate mesenchymal stem cells into neural stem cells.Methods Seven-week-old Wistar rats were selected.Zhenbao pills were orally administered and serum was taken to prepare drug culture medium.Adipose was collected during operation,adipose mesenchymal stem cells were extracted,isolated and purified,and then identified.Subsequently,adipose-derived mesenchymal stem cells were stimulated with different concentration of drug serum at different time,and compared with the differentiation induced by BHA chemical method.The expression of NSE(neuron-specific enolase)was detected by RT-PCR and Western blot at the level of gene and protein,respectively.Results1.The results of flow cytometry: Flow cytometry showed that the cells were strongly positive for CD-49 d,CD-13,CD-59 and weakly positive for CD106.This is the characteristic of adipose-derived mesenchymal stem cells.2.RT-PCR results : Adipose-derived mesenchymal stem cells stimulated by drug serum could increase the expression of neuron-specific protein NSE.There were significant differences between adipose-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells induced by untreated serum.3.Western blot results: Adipose-derived mesenchymal stem cells stimulated by drug serum can express the specific protein NSE of neuron cells.It was also found that the expression of specific protein NSE increased with the increase and extension of stimulation time.Conclusions1.Mongolian medicine Zhenbao Pills can induce adipose-derived mesenchymal stem cells to differentiate into neuron-like cells.2.In this induced differentiation experiment,the induction effect of Mongolian medicine Zhenbao Pill was stronger than that of butyl hydroxy anisole.3.When the serum concentration of Zhenbao pill was 10%,the effect of Zhenbao Pill on the differentiation of adipose mesenchymal stem cells into neurons was the strongest. |
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