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作 者:丁振禹[1] 赵翠香[2] 王凤斌[2] 刘冰[2] 谭春艳[2] 王亚南[2]
机构地区:[1]上海交通大学医学院,上海200025 [2]潍坊医学院
出 处:《中华行为医学与脑科学杂志》2013年第9期791-793,共3页Chinese Journal of Behavioral Medicine and Brain Science
基 金:山东省科技发展计划项目(2007GG30002007)
摘 要:目的观察植物雌激素三羟基异黄酮(genistein,GST)对APP695基因转染PCI2细胞Bax和Bcl-2蛋白表达的影响。方法用pIRES2-EGFP空载体和pIRES2-EGFP/APP695MT表达载体转染正常PCI2细胞,将细胞分为正常对照组、空载转染组、APP695转染组和GST干预组(5tLmol/L、15Ixmol/L)。采用免疫细胞化学SABC法和免疫印迹法检测凋亡相关蛋白Bax和Bcl-2在PCI2细胞中的表达。结果免疫细胞化学SABC法结果显示,与正常对照组比较,APP695转染组PCI2细胞Bax蛋白细胞阳性表达率显著升高[(78.02±1.26)%,P〈0.01)],Bcl-2蛋白细胞阳性表达率则明显降低[(15.40±0.72)%,P〈0.01)];15~mol/LGST组与APP695转染组比较,Bax蛋白的阳性细胞表达率显著降低[(22.35±0.49)%,P〈0.01)],而Bcl-2蛋白的阳性表达率明显增强[(68.11±0.24)%,P〈0.01)]。Westernblot结果表明,APP695转染组与对照组比较Bax蛋白的表达量明显增多,Bcl-2蛋白表达则明显降低;GST各处理组与APP695转染组比较,Bax蛋白的表达量明显降低,Bcl-2蛋白表达显著增强。结论GST能降低APP695基因转染引起的PCI2细胞内Bax蛋白表达的水平,而提高Bcl-2蛋白的表达水平,对PCI2细胞产生保护作用。Objecti±e To in±estigate the effect of genistein on the expression of Bax and Bcl-2 protein in PC12 cells transfected with mutant APP695 Type. Methods PC12 cells were transfected with pIRES2-EGFP plasmid or pIRES2-EGFP/APP695MT expression plasmid, and then di±ided into normal control group, Unladen group,APP695 transfected group,GST treatment group(5 μmol/L, 15 μmol/L). The Bax and Bcl-2 protein ex- pression in PC12 cells was measured by Immunocytochemistry SABC and Western-blot. Results Immunocytochemistry SABC showed that:compared with the normal control group, the expression of bax protein in APP695 transfected group was strongly increased( (78.02 ± 1.26) % , P 〈 0.01 ) ) and the expression of bcl-2 protein was ±ery weak oppositely ( ( 15.40 ± 0.72 ) % , P 〈 0.01 ) ). Compared with APP695 transfected group, the expression of bax protein in cells of 15umol/L GST treatment group was significantly decreased ( (22.35 ± 0.49)% , P 〈 0. 01 ) ) and the expression of bcl-2 protein in these cells was strongly increased ( (68.11 ±0.24)%, P〈0.01 ) ). West- ern-Blot showed that:compared with the normal control group,the expression of bax protein in APP695 transfected group was markedly increased and the expression of Bcl-2 protein was significantly decreased. Compared with the APP695 transfected group,the expression of Bax protein was markedly decreased and the expression of Bcl-2 pro- tein was significantly increased in the cells of the GST treatment groups. Conclusion GST can reduce the expres- sion of bax protein and the increased expression of Bcl-2 protein has protecti±e effect on PC12 cells transfected with APP696MT.
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