Abstract

ANALYZING THE EFFECT OF BETA-BOSWELLIC ACID (BBA) TO DOPAMINERGIC NEURON DIFFERENTIATION OF TRANSGENE EXPRESSING MOUSE EMBRYONIC STEM CELLS

Mojtaba Safi, Hossein Rahmani, Sajjad Biglari*, Adeleh Divsalar

Abstract

Introduction: Parkinson's disease is a degenerative neurological disorder associated with progressive loss of dopamine-secreting neurons in the direction of the Substantia nigra and the basal ganglia. In the past, drug treatment was the only known solution for the disease. But now it has been tries to use modern medical methods and herbal extracts to reduce the effects of chemical drugs. Given the above, the main aim of the present study was to investigate the effect of beta-boswellic acid (BBA) for dopaminergic production of produce embryonic stem cells expressing the foreign genes using lentivirus vectors in Parkinson's disease. Method: In this study, using the gene transfer techniques based on lentivirus vectors, Nurr1 genes (A transcription factor involved in the development of DAn and GPX1 (An antioxidant enzyme) are transferred to the R1 murine embryonic stem cells and the permanent expression of these genes in these cells is confirmed. For the differentiation of murine embryonic stem cells expressing the nurr1 and gpx1 genes to Dopaminergic neurons with the function the cell experiments are used and the making EB protocol was used to distinguish the embryonic stem cell to dopaminergic neurons. Then at the end of the third, fourth and fifth phases of differentiation we analyzed the expression of markers of neural precursor cells ( Nestin ) and the markers of dopaminergic neurons (Tau neuron TH, Nurr1, Pitx3 ) by Real time PCR quantitative techniques. At the third and fifth phases the expression of specific markers was studied. Results: The results showed that the stem cells infected with lentivirus maintained their pluripotency characteristics after receiving Nurr1 and GPX1 genes. Also in infected cells, foreign genes are expressed at the level of mRNA and also the results indicated that the simultaneous expression of Nurr1 and GPX1 along with BBA treatment had the greatest effect in increasing the neural and neuronal markers and eventually in the expression of specific markers of dopaminergic neurons. Conclusion: The results suggest that the BBA has a positive impact on the differentiation of stem cells from dopaminergic cells and the markers of dopaminergic neurons in the groups under study was expressed and in the group of foreign genes treated with BBA the highest amount of these markers was observed. Also the therapy cell as a treatment to replace lost neurons can be a permanent and effective treatment for the disease.

Keywords: beta-boswellic acid, differentiating murine embryonic stem cells, dopaminergic, lentivirus vectors, Parkinson.