Mechanism of Tumor Necrosis Factor Alpha Up-regulating RH Protein C Expression in Human Brain Microvascular Endothelial Cells
Objective This article aims to investigate the regulation of RH protein C(RHCG)expression in human brain mi-crovascular endothelial cells(HBMEC)by pro-inflammatory factor tumor necrosis factor α(TNF-α),which plays a strong role in hepatic encephalopathy,and its potential mechanism.Methods HBMEC was resuscitated,passed for 5 generations,cultured in in-cubators with a suitable concentration of perforated plates,and co-cultured with TNF-α(0.1 μg/mL endothelial cell medium ECM).Reverse transcription PCR(RT-PCR)and Western Blot were used to detect the expression levels of RhCG mRNA and pro-tein in HBMEC with the extension of TNF-α action time,and the change trend was statistically analyzed.According to the above re-sults,the time point with the highest expression level was selected and treated with PKC inhibitor(Safingol),and divided into the following groups:(1)group treated with TNF-α for 0 hours;(2)group treated with TNF-α for 24 hours;(3)Safingol alone treat-ment group;(4)TNF-α+Safingol inhibitor group:HBMEC cells were treated with Safingol for 1 hour and then stimulated with TNF-α for 24 hours.Western Blot was used to detect the expression level of RhCG protein in HBMEC,and the change trend of RhCG pro-tein in different groups was analyzed.Results The expression of RhCG protein increased with the prolongation of TNF-α,and the expression level was the highest at 24 hours.The expression of RhCG mRNA increased with the extension of TNF-α action time,and the expression level was the highest at 24 hours.After the addition of PKC-α specific inhibitor Safingol,the expression of RhCG pro-tein decreased,and the difference was statistically significant.Conclusion Tumor necrosis factor TNF-α can up-regulate the ex-pression of RhCG mRNA and protein in HBMEC cells.PKC-α is involved in the regulation of TNF-α up-regulation of RhCG protein expression in HBMEC.
NETL
NSTL
万方数据
