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Objective To investigate the cellular receptor pathway and the intracellular signaling of advanced glycation end products(AGE)-induced inflammatory reaction in monocytes. Methods Human peripheral monocytes were isolated from healthy volunteers. Cells were incubated with AGE modified by the addition of human serum albumin (AGE-HSA) either with pretreatment or no pretreatment of anti-AGE receptor (RAGE) IgG, NADPH oxidase inhibitor (apocynin)or a specific inhibitor of p38(SB 203580). The levels of interleukin-1?(IL-1?)and tumor necrosis factor-?(TNF-?) in the supernatants were assayed with enzyme-linked immunoadsorbent assay (ELISA). Reactive oxygen species (ROS) production was determined by MCLA chemiluminescence. Nuclear factor-?B translocation was assayed by immunochemical staining with anti-NF-?B/p65 and electrophoretic mobility shift assay(EMSA). Results AGE-HSA was found to induce activation of NF-?B, increase levels of IL-1? and TNF-? in the supernatants, and enhance production of ROS by monocytes. Pre-treatment of cells with anti-RAGE IgG or apocynin inhibited AGE-HSA to induce NF-?B translocation and IL-1? or TNF-? production. AGE stimulated ROS production could also be blocked by pre-treatment of cultured cells with anti-RAGE IgG or apocynin. Pre-treatment of cultured cells with SB 203580 inhibited both NF-?B activation and cytokines production, but showed no significant effect the cells to produce ROS. Conclusion AGE-HSA could induce IL-1? and TNF-? release as well as ROS production in human monocytes via a pathway mediated by RAGE. Activation of NADPH oxidase may be the upstream of the intracellular pathway. AGE-induced cytokines production was p38 pathway-dependent.
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Objective A close relationship between atherosclerosis and plasma levels of advanced oxidation protein products (AOPP) has been demonstrated in patients with chronic renal failure. The present study was performed to investigate the effect of AOPP on human endothelial cells. Methods Human umbilical vein endothelial cell line ECV304 was incubated with different concentrations of AOPP-BSA, which prepared by combining BSA with HOCl at different molar ratios. Cell viability was measured by MTT assay. Intracellular production of reactive oxygen species (ROS) was evaluated kinetically using VICTOR Wallac 1420 mutilabel counter. Results AOPP-BSA decreased endothelial cell viability, which was dependent on the molar ratio of BSA/HOCl and the concentration of AOPP-BSA. AOPP-BSA also enhanced the intracellular ROS production in ECV304. AOPP-induced ROS production was correlated with the molar ratio of BSA/HOCl and the concentration of AOPP-BSA. Pretreatment of cells with a small molecular glutathione peroxidase mimic (ebselen) reduced AOPP-induced ROS production by 53% with preservation of cell viability. Conclusion AOPP decreased endothelial cell viability via induction of oxidative stress.
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[Objective] To investigate the influence of serum containing Danggui Buxue Decoction (DBD) on chemotaxis of oxidized low-density lipoproteins ( OxLDLs) for monocytes ( MC). [ Methods ] Six New Zealand rabbits were randomized to DBD group, Radix Angelicae Sinensis (RAS) group and Radix Astragali seu Hedysari (RASH) group. The rabbits were treated by gastric infusion of the above drugs respectively, bid for three days. On the third day, the serum containing drugs was obtained by heart blood sampling one hour after administration. Free chemotaxis group (normal control group) and OxLDLs group ( model group) were established for control. The number and figure of OxLDLs - chemotactic MC were observed by micropore filter method with chemotactic chamber. [Results] OxLDLs -chemotactic MC number in DBD group were lower than those in model group (P
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Objective To test the hypothesis that advanced glycosylation end products(AGEs) increase cellular inflammation in atherosclerotic plaques. Methods Fifty rabbits were randomly divided into five groups. Hypercholesterolemic (0.5% cholesterol in diet) rabbits received repeated intravenous injection of either AGEs modified rabbit serum albumin (AGEs-RSA ) (group A) or unmodified RSA (group B) for 10 weeks. Rabbits treated with either hypercholesterolemic diet (group C)or with a normal diet(group D) or with a normal diet, and intravenous injection of AGEs-RSA (group E) were served as controls. Aortas were harvested at the 10th week, and lipid deposition was quantitated by oil red 0 staining. Macrophage (RAM-11 positive cells) and T lymphocyte (CD43 positive cells) infiltration, smooth muscle cell(?-actin positive cells) migration and proliferation were determined by using immunohistochemical staining and image-analysis techniques. Results Atherosclerotic plaques could be found in animals fed with hypercholesterolemic diet.Lipid deposition in plaque was significantly higher in group A (71.86%?8.3%) than those in group B (53.76%?3.72%)and group C (56.67%?9.2%). Infiltrations of macrophage[ (23.1?8.5)/0.01 mm2]and T lymphocyte[ (15.1 ? 3.8)/0.01 mm2]as well as migration and proliferation of smooth muscle cell [ (19.2?5.7)/0,01 mm2] in atherosclerotic lesions were significantly increased in animals treated with hypercholesterolemic diet and received injection of AGEs-RSA (group A) when compared with group B [macrophage (14.4? 5.9)70.01 mm2; T lymphocyte (9.1?2.6)/0.01 mm2; smooth muscle cell (12.9?3.8)/0.01 mm2]and group C[macrophage (15.4?4.4)/0.01 mm2; T lymphocyte (10.5?2.2)/0.01 mm2, smooth muscle cell (13.8?3.9)/0.01 mm2]. Neither plaque nor a cellular inflammation was found in animals fed with normal diet (group D)and in those received repeated injections of AGEs-RSA (group E). Conclusion AGEs increase cellular inflammation in atherosclerotic plaques and may accelerate formation of atherosclerosis in AGEs associated diseases.
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Objective To investigate the protective effect of extract of Picrhoriza scrophulariflora on renal ischemia/reperfusion injury(I/R). Methods Male SD rats were randomly divided into sham-operated group (n=5), I/R group(n=8), I/R+ extracts of Picrhoriza scrophulariflora groups (each group had 8 rats). Acute renal ischemia/reperfusion injury in rats was reproduced by removing the right kidney and clamping the left renal artery with a non-traumatic vascular clamp for 60min followed by reperfusion for 72h. Serum creatinine, urine creatinine and renal pathological changes, were compared between I/R and I/R+ extracts of Picrhoriza scrophulariflora groups, malondialdehyde (MDA) and glutathione peroxidase (GSHPx). Results CCr levels were significant decreased after renal I/R injury. Kidneys of animal with I/R injury displayed significant pathological changes. 72h after ischemia, Ccr and serum GSHPx of animals of the I/R+ extracts of Picrhoriza scrophulariflora group were significantly higher than those of I/R group(P
