Screening of differential proteins binding to Nox1 promoter in A549 cell model of inflammation and oxidative stress / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the regulatory proteins involved in Nox1 promoter activation in a cell model of inflammation and oxidative stress.</p><p><b>METHODS</b>A cell model of inflammation and oxidative stress was established by stimulating A549 cells with tumor necrosis factor-α (TNF-α). The differential proteins binding to Nox1 promoter were screened by DNA pull-down and the binding proteins were separated by 2D electrophoresis and selected according to the their differential expression levels (with over 1.5-fold changes relative to the control level). The screened proteins were finally identified by MALDI-TOF/TOF-MS.</p><p><b>RESULTS</b>Seven differentially expressed protein spots (all upregulated in the cell model) were obtained, among which GLE1, DDX19A, KRT1 and KRT10 were identified by mass spectrometry.</p><p><b>CONCLUSION</b>GLE1, DDX19A, KRT1 and KRT10 participate in the activation of Nox1 promoter in TNF-α-induced A549 cells, and this result provides new insights into the biological roles of the regulatory proteins of Nox1 promoter in inflammation and oxidative stress.</p>

Black rice anthocyanidins prevent retinal photochemical damage via involvement of the AP-1/NF-kappaB/Caspase-1 pathway in Sprague-Dawley Rats

The effects of black rice anthocyanidins (BRACs) on retinal damage induced by photochemical stress are not well known. In the present study, Sprague-Dawley rats were fed AIN-93M for 1 week, after which 80 rats were randomly divided into two groups and treated with (n = 40) or without BRACs (n = 40) for 15 days, respectively. After treatment, both groups were exposed to fluorescent light (3,000 +/- 200 lux; 25degrees C), and the protective effect of dietary BRACs were evaluated afterwards. Our results showed that dietary BRACs effectively prevented retinal photochemical damage and inhibited the retinal cells apoptosis induced by fluorescent light (p < 0.05). Moreover, dietary BRACs inhibited expression of AP-1 (c-fos/c-jun subunits), up-regulated NF-kappaB (p65) expression and phosphorylation of IkappaB-alpha, and decreased Caspase-1 expression (p < 0.05). These results suggest that BRACs improve retinal damage produced by photochemical stress in rats via AP-1/NF-kappaB/Caspase-1 apoptotic mechanisms.

Study on changes of protein phosphorylation of p65, IkappaBalpha and IkappaBepsilon in lymphocytes of rats in progress of aging and interventional effect of Epimedium flavonoids / 中国中药杂志

<p><b>OBJECTIVE</b>To study the characteristics of protein phosphorylation of p65, IkappaBalpha and IkappaBepsilon in lymphocytes of rats in the progress of aging and the the interventional effect of Epimedium flavonoids (EF).</p><p><b>METHOD</b>We chose the lymphocytes derived from SD rat spleen. We divided the SD rats into five groups i. e. 4 months (4 m), 27 months (27 m), the 27 m EF treated group (27 m + EF), and we also observed the whole interventional effect of PDTC (a NF-kappaB inhibitor) on old rat groups (27 m PDTC, 27 m PDTC + EF ) when IkappaBepsilon, IkappaBalpha were detected. Through the western-blotting analyses, we studied the entire characteristics and distinctions of phosphorylation expression in molecules related to NF-kappaB signal transduction pathway p65, IkappaBepsilon and IkappaBalpha in lymphocytes across the age spectrum of rats in aging. We also observed the whole interventional effect of EF on lymphocytes of old rats.</p><p><b>RESULT</b>With the increasing of age, the mean level of phosphorylation expressions of p65, IkappaBalpha and IkappaBepsilon in rat spleen lymphocytes decreased obviously, When inhibited NF-kappaB by PDTC, there was decreased evidently, while PDTC + EF can active NF-kappaB family and the above molecules were increased to a certain extent.</p><p><b>CONCLUSION</b>The phosphorylation expressions of p65, IkappaBalpha and IkappaBepsilon in rat spleen lymphocytes were not enouphe, EF have a strong effect to upregulated the expression of them during aging.</p>

Effects of NKG2D and its ligands RAE-1 and H60 on graft-versus-tumor response / 中国实验血液学杂志

The study was purposed to explore the effects of NKG2D receptor and its ligands RAE-1 and H60 on graft-versus-tumor (GVT) response induced by MHC haploidentical bone marrow/spleen cell transplantation. Female (BALB/c x C57BL/6) F1 mice (CB6F1, H-2K(b/d)) inoculated with H22 cells to develop a solid tumor model were the recipients, and bone marrow mixed with spleen cells of the healthy male C57BL/6 (H-2K(b)) mice were the donor cells. GVT response was observed after transplantation that from donor cells T and NK cells were purged with anti-CD3 and anti-NK monoclonal antibody, and the NKG2D receptor was blocked with anti-NKG2D monoclonal antibody, the expression levels of RAE-1 and H60 mRNA in tumor tissue were measured by means of semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) at different time points after transplantation. The results showed that the GVT response of transplantation was reduced after in vitro depletion of T and NK cells or blocking NKG2D receptor in donor cells of the graft, the expression levels of RAE-1 and H60 mRNA in tumor tissue increased after transplantation of haploidential bone marrow mixed with spleen cells. It is concluded that NKG2D and its ligands RAE-1 and H60 may play important roles in GVT response.