Armaghane Danesh Bimonthly Journal (Sep 2023)
The Ability of Pregnenolone in Proliferation of Mouse Neural Stem Cells and Reduction of Inflammatory and Oxidant Markers after Induction of Inflammation with Lipopolysaccharide in Vitro
Abstract
Background & aim: Pregnenolone acts as a precursor to other steroid hormones and exerts its effect as an anti-inflammatory molecule to maintain immune homeostasis in various inflammatory conditions. In these diseases, a decrease in the level ofP has been observed, which emphasizes its role in neuroprotection and nerve regeneration and its anti-inflammatory role. Accordingly, the purpose of the present study was to determine the ability of Pregnenolone in the proliferation of mouse neural stem cells and reduce inflammatory and oxidant markers. of inducing inflammation with lipopolysaccharide in laboratory conditions. Methods: In the present experimental study conducted at Yasuj University of Medical Sciences, neural stem cells (NSCs) were isolated from the embryonic cortex of E14 mice with standard protocol and incubated for 5 days. Subsequently, neurosphere formation and propagation for second passage the survival of the cells was done after pregnenolone combined treatment with lipopolysaccharide (LPS) inflammatory model. The number of neurospheres and cells derived from neurospheres were counted after 5 days of incubation in the inflammatory model. The supernatant of the cells was removed and the levels of oxidant and antioxidant markers MDA, NO FRAP, and inflammatory markers IL6 and TNFα were measured by ELISA method. Data were analyzed by one-way variance statistical method and Tukey's post hoc test. Results: The results indicated that pregnenolone with its effect on inflammatory factors could increase the proliferation of neural stem cells in conditions of inflammation and the greatest effect was observed in the group treated with 10 μM dose of pregnenolone with an increase of 68% compared to the LPS group. On the other hand, it caused a decrease in the inflammatory factors TNF-α (12%) and IL-6 (30%) and oxidative stress factors including NO (38%) and MDA (20%) compared to the LPS group, as well as a significant increase FRAP was an antioxidant marker (P<0.0001) in the model of inflammation caused by LPS in the culture medium of mouse neural stem cells. Conclusion: The results of the present study indicated that Pregnenolone, by affecting inflammatory factors, increased the proliferation of neural stem cells in the conditions of inflammation, and it was as well able to reduce the amount of inflammatory and oxidant markers in the inflammatory model of the culture medium.
