中国生物工程杂志

Objective: To study the role of protein arginine methyltransferase 5 (Prmt5) in cerebral vascular development and homeostasis maintenance in mice, and to investigate the effect of Prmt5 specific knockout on the central nervous system. Methods: We crossed Prmt5^fl/fl mice with SP-A Cre transgenic mice that express Cre recombinase in cerebrovascular endothelial to generate cerebrovascular endothelial cell-specific Prmt5 knockout mice. H-E staining and immunostaining were performed to observe the vascular structures of control and Prmt5^fl/fl mutant mice. Laser speckle contrast imaging was used to detect cerebral blood flow in control and mutant mice. Sulfo-NHS-Biotin was intraperitoneally injected into control and mutant mice to examine the blood brain barrier(BBB) integrity. The expression levels of astrocyte glial fibrillary acidic protein (GFAP), S100 calcium-binding protein β (S100β), complement C3 (C3), C1q, tumor necrosis factor alpha(TNF-α) and Interleukin-1 beta(IL-1β) were detected by immunofluorescence and Western blot to evaluate the activation level of astrocytes in cortex, thalamus and cerebellum of knockout mice and control mice. Furthermore, activators of astrocytes, such as C1q, TNF-α, IL-1β and other cytokines, were also detected by real-time PCR. Results: We found that cerebrovascular endothelial cell-specific Prmt5 knockout mice exhibited aberrant cerebrovascular structure, and increased the number of reactive astrocytes. The expression levels of TNF-α and IL-1β in the whole brain, as well as the C1q, TNF-α and IL-1β, were all increased in Prmt5^fl/fl mutant mice. Conclusion: Prmt5 plays an essential role in the maintenance of cerebrovascular homeostasis, suggesting that it might act as a potential therapeutic target for cerebrovascular diseases.