ABSTRACT
Objective:To study interferon alpha (IFN-α) inhibition of proliferation and apoptosis induction of human brain vascular adventitial fibroblasts(HBVAFs)via IFI16.Methods:Cultured HBVAFs were treated with transfection IFI16 siRNA and/or IFN-α in vitro instantaneously.The protein and mRNA levels of IFI16,P53,P21 were measured by Western blot and Real-time PCR.MTT was used to detect the cell proliferation of the HBVAFs.Cell cycle and apoptosis were analyzed by flow cytometry.Results:IFN-α with terminal concentration of 2 000-5 000 kU/L induce significantly expression of IFI16 in HBVAFs,without any significant difference.Stimulated with 2 000 kU/L IFN-α up-regulated the expression of P53,P21 at protein and mRNA levels,and inhibited the cell proliferation and promote cells apoptosis in HBVAFs.Such effect was restrained by transfection with IFI16 siRNA into HBVAFs.Conclusion:IFN-α inhibits HBVAFs proliferation and induces apoptosis may partly relate to the increased IFI16 expression.
ABSTRACT
Objective To investigate the effect of interferon alpha ( IFN-α) on apoptosis of vascular smooth muscle cells ( VSMCs) in rats and the related mechanism. Methods The cells were divided into three group:group A, group B and group C.Group A was transfected with nonspecific siRNA, group B was intervened with IFN-α and transfected with nonspecific siRNA, and group C was intervened with IFN-α and transfected with IFI204 siRNA. All the cells were cultured for 48 h. The expression of P204 mRNA was determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).P204, RAS protein levels, and phosphorylation levels of RAF and ERK were analyzed by Western blotting. The cell apoptosis was analyzed by flow cytometry with Annexin-V FITC/PI method. Results As compared with group A, the expression of P204 mRNA and protein in group B was up-regulated (P<0.05), the cell apoptosis was increased (P<0.05), in the process of the above, the expression of RAS protein was decreased ( P<0.05) and the phosphorylation levels of RAF and ERK were dropped (P<0.05).In group C, the expression levels of P204 mRNA and protein were down-regulated (P<0.05), and cell apoptosis was decreased ( P<0.05) , the expression of RAS protein and the phosphorylation levels of RAF and ERK were increased ( P<0.05) . Conclusion P204 and RAS signal pathway participates in IFN-α regulation of apoptosis of VSMCs in rats.
ABSTRACT
Objective: To observe the effects of interferon-induced protein 204 (p204) over-expression on apoptosis, proliferation and migration of aortic vascular adventitial ifbroblast (VAFs) in experimental rats. Methods: Our research included in 3 groups: Iif204-Lv group, in whichVAFs were infected by Iif204-recombined lentivirus, Con-Lv group, in which VAFs carried the empty vector without virus, Blank control group, in which VAFs were untreated. VAFs proliferation was examined by MTT method, cell apoptosis was measured by lfow cytometry and the migration was detected by scratching assay and transwell chamber method. The mRNA and protein expressions of p204, p53 and p21were evaluated by real-time q RT-PCR and Western blot analysis respectively. Results: Compared with Con-Lv and Blank control groups, Iif204-Lv group had decreased VAFs proliferation (by OD value) at 48 hours: (0.53 ± 0.05) vs (0.66±0.03) and (0.63 ± 0.06), at 72 hours: (0.89 ± 0.06) vs (1.02 ± 0.06) and (1.01 ± 0.07); distance of cell migration (by pixel): (61.00 ± 1.83) vs (74.50 ± 6.25) and (75.50 ± 7.85); number of cell migration: (61.75 ± 10.69) vs (155.25 ± 10.21) and (153.75 ± 9.40), allP Conclusion: p204 over-expression inhibits VAFs proliferation and migration which might be partly related to the activation of p53 and p21 expression in experimental rats.