Effect of anti-Aβ42 antibody on Aβ42 stimulating inflammatory factors production in microglia
10.3760/cma.j.issn.0254-9026.2010.09.021
- VernacularTitle:抗淀粉样蛋白42抗体对小胶质细胞释放炎性介质的影响
- Author:
Jianwei MO
;
Yunyong FANG
;
Dongfeng LI
;
Jinhai DUAN
- Publication Type:Journal Article
- Keywords:
Alzheimer's disease;
Amyloid beta-protein;
Microglia;
Interleukins;
Nitric oxide
- From:
Chinese Journal of Geriatrics
2010;29(9):779-782
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the effect of β-amyloid42 (Aβ42) on stimulating the inflammatory factors production by BV-2 microglia, including interleukin-1β (IL-1β), IL-10 and nitric oxide (NO), and to contrast the inhibitory action of anti-Aβ42 antibody in serum of Alzheimer's patients and the artificially synthesized anti-Aβ42 antibody. Methods The anti-Aβ42 antibodies were extracted from the serum of Alzheimer's patients. And the BV-2 microglia cells in murine were cultured as in vitro cell model. The cells were stimulated by Aβ42 and the two different anti-Aβ42antibodies to analyze the impact of stimulants on the cell activity. The Aβ42 of 5 μmol/L was added to the culture separately or in mixture with each of the two different anti-Aβ42 antibodies. Each antibody was mixed with Aβ42 of 5 μmol/L at final anti-Aβ42 antibody titre of 5 μg/ml, 1 μg/ml and 0.2 μg/ml,respectively. Then clear supernatant was collected from each tube respectively at 6 h, 12 h, and 24 h after culture, and the concentrations of IL-1β, IL-10 and NO were determined. Results The Aβ42,artificially synthesized anti-Aβ42 antibody and anti-Aβ42 antibody from Alzheimer's patients had no effects on the activities of BV-2 cells, the cell survival rates were (98. 6±5.8)%, (101.9±2.8)%and (98. 4±6.0)%, with no significant differences as compared with normal control group (F=0. 407, P＞0. 05). The inflammatory factors releasing from BV-2 cells stimulated by Aβ42 reached the peak level at 12 h, the concentrations of IL-1β, IL-10 and NO were (69.0±12.7) pg/ml, (24.1 ±4. 0) pg/ml and (128. 2±8. 7) μmol/L, the concentrations of IL-1β and NO were significantly higher at 12 h than at 6 h and 24 h (F= 15. 470 and 242. 107, P＜0.05), there was no significant difference in the concentration of IL-10 among 12 h, 6 h and 24 h (F=1. 852, P＞0.05). The two different antiAβ42 antibodies of different titre remarkably inhibited Aβ42 stimulated BV-2 cells to release inflammatory factors. At high titre of 5 μg/ml, the two different antibodies showed no significant difference in the inhibitory effects (P＞0.05), while at the titre of 0. 2 μg/ml, anti-Aβ42 antibody from Alzheimer's patients showed a significantly lower inhibitory effect than artificially synthesized antibodies, the concentrations of NO were (35.4 ± 2. 5) μoml/L and ( 19. 2 ± 3.3) μoml/L,respectively (P ＜ 0.05). Conclusions The Aβ42 can stimulate BV-2 microglia cells to release inflammatory factors. Anti-Aβ42 antibody from Alzheimer's patients has a lower inhibitory effect on Aβ42 in stimulating microglia to release inflammatory factors.
