To explore the effect of CtBP-Interacting protein (CtIP) on oxidative damage of cerebral endothelia cells and its mechanism. Cerebral endothelia cells were stimulated by TBHP to induce oxidative damage. The cell line of CtIP gene were prepared by over-expression and interfering lentivirus technology. Cell damage was detected by immunofluorescence assay of cysteinyl aspartate specific proteinase-3 (Caspase-3). The expression of CtIP and Caspase-3 protein was detected by Western blotting, and the related genes of CtIP signaling pathway were detected by Realtime RT-PCR. The results of immunofluorescence and Western blotting showed that overexpression of CtIP inhibited the Caspase-3 expression reducing to 1/3 level compared with normal cultured cerebral endothelia cells. Interfering with CtIP expression resulted in the Caspase-3 expression increased significantly to 4/5 level compared with normal cultured cerebral endothelia cells and cerebral endothelia cells were damaged more severely. Realtime RT-PCR data showed that expression of CtIP significantly increased the expression of BRCA1 and ZBRK1 genes, but inhibited the expression of p21 gene. It is confirmed that CtIP gene has the significantly inhibitory effect on injured cerebral endothelia cells, and the regulatory relationship between CtIP gene and BRCA1, ZBRK1 and p21 genes in the process of injury are determined.
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http://dx.doi.org/10.3760/cma.j.cn112137-20190919-02074 | DOI Listing |
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