Empowerment of Balb/C Mouse Neuron and Glial Cells in Steroidogenesis After Activation of the SHH Signaling Pathway and Co-Treatment with Pregnenolone

Abstract

Background & aim: Steroid production has been reported in the asexual tissues of the nervous system. Stimulants are in the normal activity, function and function of the nervous system. Identifying the conduction pathways involved in glucocorticoids and enabling brain parenchymal cells can offset the balance in the active nervous system at old ages when the body is depleted. Therefore, in this study, by increasing the activity of sonic hedgehog (SHH) pathway attempted by purmorphamine and its capacity by Gant 61, the effect of this pathway on steroid process in culture medium of glial neurons is evaluated. Methods:The present experimental study was conducted in 2021. First, neuronal stem cells were obtained from the cortex of a 14-day-old embryonic mice by standard methods. Survival of neuronal stem cells after treatment with 5 μM pregnenolone with different concentrations of purmorphamine (1,2,5, 10 and 20) and Gant 61 was performed by MTT method. Then the cells were placed in a differentiation medium and after treatment with different concentrations and 5-day incubation, the surface of the cells was removed from the cell culture medium and the amount of testosterone and estrogen were measured by ELISA and HPLC. Data were analyzed using ANOVA statistical test using graph pad software. Results:The survival data of the groups indicated an increase in survival after treatment with purmorphamine (114.3) compared to the Gant 61 group (63.7 Pg) (p≤0.5). Progesterone data in the supernatant of glial neurons showed that purmorphamine groups (287.2 Pg) had a significant increase compared to control (88.28) and Gant 61 (40.5 Pg) groups (p≤ 0.001). Also, testosterone data show that purmorphamine groups (73.8 Pg) in both ELISA and HPLC methods have a significant increase compared to the control (153.8 Pg) and Gant61 (52.92 Pg) groups (p≤ 0.0001). Also, pregnenolone group (236.5 Pg) showed a significant increase compared to Gant61 (40.5 Pg) group (p≤ 0.05). Analysis of estrogen data by HPLC method showed that there was a significant increase in estrogen production in the purmorphamine groups (331.2 Pg) compared to the control (42.11 Pg) and Gant61 (42.11 Pg) groups (p≤ 0.0001). Conclusion:The data from this study indicated that the induction of the shh pathway by purrmorphamine increased the production of steroid hormones (estrogen-progesterone and testosterone) by glial-neuronal differentiating cells, which inhibition of this pathway had the opposite effect. The present study concluded that induction of the shh pathway can lead to the production of steroids.