Aβ不同片段肽疫苗免疫鼠后血清对Aβ_(42)诱导细胞毒性作用的影响  

The Effects of Immunized Rat Sera of AβDifferent Peptide Vaccine on Cytotoxicity Induced by Amyloid βPeptide

作　　者：李国营[1] 胡金家[1] 林贤[1] 徐杰[1] 汪华侨[1] 姚志彬[1] 

机构地区：[1]中山大学基础医学院人体解剖学教研室脑研究室,广东广州510080

出　　处：《中山大学学报（医学科学版）》2004年第2期102-104,109,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金资助重点项目(20013137);广东省自然科学基金(010698)

摘　　要：【目的】探讨Aβ42及其亚单位疫苗免疫鼠后其血清对Aβ42诱导的细胞毒性作用的影响,为其体内应用提供依据。【方法】①用间接ELISA法测定Aβ42和K7Aβ36-42疫苗接种鼠后的血清中抗体滴度;②将不同浓度的Aβ42和K7Aβ36-42加入培养有PC12细胞的96孔培养板中,检测其对PC12细胞的毒性作用;③用不同浓度的Aβ42及K7Aβ36-42疫苗免疫鼠血清加入含Aβ42的培养基中,与PC12一起孵育培养,7d后用MTT法和酶标仪测定存活细胞的A值。【结果】①Aβ42和K7Aβ36-42疫苗接种后血清的平均抗体滴度分别为1∶6800和1∶4600;②10mg/L和20mg/LAβ42组细胞的存活率与空白组相比差异有显著性(P<0.01);③各浓度K7Aβ36-42组与空白组相比差异均无显著性(P>0.05)。在浓度为20mg/LAβ42的培养基中加入Aβ42疫苗或K7Aβ36-42疫苗免疫鼠血清,各浓度的免疫鼠血清组和阳性对照组相比差异均有显著性(P<0.05或P<0.01)。【结论】Aβ42疫苗和Aβ亚单位疫苗免疫鼠后,其血清均能在体外抑制Aβ42对细胞产生的毒性作用。To explore on the effect of Aβ42 vaccine and Aβ42 subunit vaccine immunized rat sera on counteracting cytotoxicity of amyloid βpeptide to PC12 cells in vitro. ①The titer of antibody against Aβ42 of rat serum was measured by indirect ELISA respectively after the inoculation with Aβ42 and K7Aβ36 42 vaccine ②Aβ42 or K7Aβ36 42 of different concentrations was added to culture medium with PC12 cells to measure their cytotoxicity. ③Anti Aβ42 or anti K7Aβ36 42 vaccine immunized rat sera was added to culture medium with PC12 cells respectively,then incubated with 20mg/L Aβ42 preparation. Cell viability rate in all groups was tested by MTT assay after seven days. The average titer of antibody of rat serum was 1∶6 800 and 1∶4 600 respectively after inoculation with Aβ42 or K7Aβ36 42 vaccine. With the concentration of Aβ42 increased , the viability rate of cells decreased. Compared with blank group, cell viability rates were significantly different between 10 mg/L Aβ42 and 20 mg/L Aβ42 groups (P >0.01 ),but all K7Aβ36 42 groups of different concentration have no statistical difference compared with blank group (P >0.05). Aβ42 vaccine or K7Aβ36 42 vaccine immunized rat sera was added to culture medium with 20mg/L Aβ42. Compared with the positive control group, cell viability rates were significantly different among all groups with different concentrations of anti Aβ42 or anti K7Aβ36 42 rat serum(P< 0.05 or P< 0.01). [Conclusion]Both anti Aβ42 and anti Aβsubunit sera can counteract cytotoxicity of Aβ42 in vitro.

关 键 词：β淀粉样蛋白 疫苗 免疫血清 细胞毒性 老年性痴呆 细胞毒性 间接ELISA法 老年斑 

分 类 号：R749.16[医药卫生—神经病学与精神病学]