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Aβ不同片段肽疫苗免疫鼠后血清对Aβ42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

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【作者】 李国营; 胡金家; 林贤; 徐杰; 汪华侨; 姚志彬;

【Author】 LI Guo ying,HU Jin jia,LIN Xian,XU Jie, WANG Hua qiao,YAO Zhi bing (Department of Anatomy and Brain Research, Preclinical Medicine School, SUN Yat sen University,Guangzhou 510080, China)

【机构】 中山大学基础医学院人体解剖学教研室脑研究室; 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; 广东广州510080; 广东广州510080;

【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。

【Abstract】 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.

【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)
  • 【文献出处】 中山大学学报(医学科学版) ,Journal of Sun Yat-sen University (Medical Sciences) , 编辑部邮箱 ,2004年02期
  • 【分类号】R392.33
  • 【被引频次】13
  • 【下载频次】175
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