中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2010年
9期
779-782
,共4页
莫建伟%方运勇%李东风%段金海
莫建偉%方運勇%李東風%段金海
막건위%방운용%리동풍%단금해
阿尔茨海默%淀粉样β蛋白%小神经胶质细胞%白细胞介素类%一氧化氮
阿爾茨海默%澱粉樣β蛋白%小神經膠質細胞%白細胞介素類%一氧化氮
아이자해묵%정분양β단백%소신경효질세포%백세포개소류%일양화담
Alzheimer's disease%Amyloid beta-protein%Microglia%Interleukins%Nitric oxide
目的 观察淀粉样蛋白42(Aβ42)刺激BV-2小胶质细胞释放炎性介质白介素-1β(IL-1β)、白介素-10(IL-10)和一氧化氮(NO)的作用,以及对抗Aβ42抗体与人工合成抗Aβ42抗体对Aβ42刺激小胶质细胞释放炎性介质的抑制作用.方法 采集AD患者血清制备提纯抗Aβ42抗体,应用小鼠BV-2小胶质细胞作为体外细胞模型.分别用Aβ42、两种不同抗Aβ42抗体刺激细胞,分析刺激物对细胞活性的影响.将5 μoml/L Aβ42单独加入,以及分别与不同滴度的AD患者血清提纯的抗Aβ42抗体及相应滴度的人工合成抗Aβ42抗体(抗体滴度依次为5 μg/ml,1μg/ml,0.2 μg/ml)充分混合后加入体外细胞模型培养,于培养后6 h、12 h及24 h提取细胞上清液,测定IL-1β,IL-10,NO浓度.结果 Aβ42,人工合成和AD患者血清提纯的抗Aβ42抗体对BV-2细胞活性无影响,细胞存活率分别为(98.6±5.8)%,(101.9±2.8)%和(98.4±6.0)%,与正常对照组比较差异无统计学意义(F=0.407,P>0.05).Aβ42刺激BV-2细胞分泌炎性因子在12 h达高峰,IL-1β和IL-10浓度分别为(69.0±12.7)pg/ml和(24.1±4.0)pg/ml,NO浓度为(128.2±8.7)μmol/L,IL-1β和NO浓度均明显高于6 h及24 h(F=15.470,242.107;P<0.05),IL-10浓度与6 h及24 h比较差异无统计学意义(F=1.852,P>0.05).不同滴度的两种抗体均能明显抑制Aβ42刺激BV-2细胞分泌炎性因子.其中,同样是高浓度(5μg/ml)情况下,两种抗体的抑制作用差异无统计学意义(P>0.05);而抗体浓度均减低到0.2μg/ml时,AD患者血清提纯的抗Aβ42抗体对Aβ42刺激小胶质细胞释放炎性因子NO的抑制作用明显低于人工合成的抗Aβ42抗体[NO的浓度分别为(35.4±2.5)μoml/L和(19.2±3.3)μoml/L,P<0.05].结论 Aβ42存在刺激BV-2小胶质细胞释放炎性因子的作用;AD患者血清提纯的抗Aβ42抗体对Aβ42刺激小胶质细胞释放炎性介质的抑制作用下降.
目的 觀察澱粉樣蛋白42(Aβ42)刺激BV-2小膠質細胞釋放炎性介質白介素-1β(IL-1β)、白介素-10(IL-10)和一氧化氮(NO)的作用,以及對抗Aβ42抗體與人工閤成抗Aβ42抗體對Aβ42刺激小膠質細胞釋放炎性介質的抑製作用.方法 採集AD患者血清製備提純抗Aβ42抗體,應用小鼠BV-2小膠質細胞作為體外細胞模型.分彆用Aβ42、兩種不同抗Aβ42抗體刺激細胞,分析刺激物對細胞活性的影響.將5 μoml/L Aβ42單獨加入,以及分彆與不同滴度的AD患者血清提純的抗Aβ42抗體及相應滴度的人工閤成抗Aβ42抗體(抗體滴度依次為5 μg/ml,1μg/ml,0.2 μg/ml)充分混閤後加入體外細胞模型培養,于培養後6 h、12 h及24 h提取細胞上清液,測定IL-1β,IL-10,NO濃度.結果 Aβ42,人工閤成和AD患者血清提純的抗Aβ42抗體對BV-2細胞活性無影響,細胞存活率分彆為(98.6±5.8)%,(101.9±2.8)%和(98.4±6.0)%,與正常對照組比較差異無統計學意義(F=0.407,P>0.05).Aβ42刺激BV-2細胞分泌炎性因子在12 h達高峰,IL-1β和IL-10濃度分彆為(69.0±12.7)pg/ml和(24.1±4.0)pg/ml,NO濃度為(128.2±8.7)μmol/L,IL-1β和NO濃度均明顯高于6 h及24 h(F=15.470,242.107;P<0.05),IL-10濃度與6 h及24 h比較差異無統計學意義(F=1.852,P>0.05).不同滴度的兩種抗體均能明顯抑製Aβ42刺激BV-2細胞分泌炎性因子.其中,同樣是高濃度(5μg/ml)情況下,兩種抗體的抑製作用差異無統計學意義(P>0.05);而抗體濃度均減低到0.2μg/ml時,AD患者血清提純的抗Aβ42抗體對Aβ42刺激小膠質細胞釋放炎性因子NO的抑製作用明顯低于人工閤成的抗Aβ42抗體[NO的濃度分彆為(35.4±2.5)μoml/L和(19.2±3.3)μoml/L,P<0.05].結論 Aβ42存在刺激BV-2小膠質細胞釋放炎性因子的作用;AD患者血清提純的抗Aβ42抗體對Aβ42刺激小膠質細胞釋放炎性介質的抑製作用下降.
목적 관찰정분양단백42(Aβ42)자격BV-2소효질세포석방염성개질백개소-1β(IL-1β)、백개소-10(IL-10)화일양화담(NO)적작용,이급대항Aβ42항체여인공합성항Aβ42항체대Aβ42자격소효질세포석방염성개질적억제작용.방법 채집AD환자혈청제비제순항Aβ42항체,응용소서BV-2소효질세포작위체외세포모형.분별용Aβ42、량충불동항Aβ42항체자격세포,분석자격물대세포활성적영향.장5 μoml/L Aβ42단독가입,이급분별여불동적도적AD환자혈청제순적항Aβ42항체급상응적도적인공합성항Aβ42항체(항체적도의차위5 μg/ml,1μg/ml,0.2 μg/ml)충분혼합후가입체외세포모형배양,우배양후6 h、12 h급24 h제취세포상청액,측정IL-1β,IL-10,NO농도.결과 Aβ42,인공합성화AD환자혈청제순적항Aβ42항체대BV-2세포활성무영향,세포존활솔분별위(98.6±5.8)%,(101.9±2.8)%화(98.4±6.0)%,여정상대조조비교차이무통계학의의(F=0.407,P>0.05).Aβ42자격BV-2세포분비염성인자재12 h체고봉,IL-1β화IL-10농도분별위(69.0±12.7)pg/ml화(24.1±4.0)pg/ml,NO농도위(128.2±8.7)μmol/L,IL-1β화NO농도균명현고우6 h급24 h(F=15.470,242.107;P<0.05),IL-10농도여6 h급24 h비교차이무통계학의의(F=1.852,P>0.05).불동적도적량충항체균능명현억제Aβ42자격BV-2세포분비염성인자.기중,동양시고농도(5μg/ml)정황하,량충항체적억제작용차이무통계학의의(P>0.05);이항체농도균감저도0.2μg/ml시,AD환자혈청제순적항Aβ42항체대Aβ42자격소효질세포석방염성인자NO적억제작용명현저우인공합성적항Aβ42항체[NO적농도분별위(35.4±2.5)μoml/L화(19.2±3.3)μoml/L,P<0.05].결론 Aβ42존재자격BV-2소효질세포석방염성인자적작용;AD환자혈청제순적항Aβ42항체대Aβ42자격소효질세포석방염성개질적억제작용하강.
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.
