中文摘要：

目的探讨小檗碱抑制Aβ产生的机理。方法稳定转染人淀粉样前体蛋白瑞典突变695的人胚胎肾293细胞（HEK293APPsw695）分别给与小檗碱（1μM,5μM,10μM和20μM）48 h、5μM小檗碱（8 h,24 h,48 h和72 h）、U0126（0.5μM）48 h以及小檗碱（5μM）＋U0126（0.5μM）48 h,然后分别通过噻唑蓝（MTT）技术和测定培养基中乳酸脱氢酶的活性研究以上各处理对HEK293APPsw695增殖和毒性的影响,通过ELISA方法检测各处理组对HEK293APPsw695产生Aβ40/42的影响,通过WB方法研究各处理组对β分泌酶和p-ERK1/2表达的影响。结果以上各处理对HEK293APPsw695细胞的增殖和毒性没有影响,小檗碱以时间和浓度依赖的方式显著抑制Aβ40/42的产生和β分泌酶的表达,以及促进p-ERK1/2的表达,U0126完全逆转小檗碱对Aβ40/42产生和β分泌酶表达的抑制,以及对p-ERK1/2表达的促进。结论小檗碱可能通过活化ERK1/2通路抑制β分泌酶的表达而抑制Aβ40/42的产生。

英文摘要：

Objective To investigate the mechanism of berberine on decreasing the production of Aβ.Method HEK293APPsw695 cells,stably transfected with APP695 containing the Swedish mutation,were either incubated with different concentrations of berberine（1 μM,5 μM,10 μM and 20 μM） for 48 h,berberine（5 μM） for 8 h,24 h,48 h and 72 h,U0126（1,4-Diamino-2,3-dicyano-1,4-bis（o-aminophenylmercapto） butadiene）（0.5 μM） or U0126（0.5 μM） ＋ berberine（5 μM） for 48h respectively.The effects of berbeine and U0126 on cell proliferation and cytotoxicity of HEK293APPsw695 cells were evaluated using MTT（3-（4,5）-dimethylthiahiazo（-z-y1）-3,5-di-phenytetrazoliumromide） assay and LDH（lactate dehydrogenase） assay.The production of Aβ40/42 and the expression of β-secretase and p-ERK1/2（extracellular regulated kinase1/2 phosphorylation） were assessed by ELISA（enzyme-linked immunosorbent assay） assay and western blot,respectively.Results Berberine could significantly decrease the production of Aβ40/42 and the expression of β-secretase and increase the expression of p-ERK1/2 in dose-dependent and time-dependent manner without inhibition of cell proliferation and cytotoxicity on HEK293APPsw695 cells.U0126 could completely alleviate the inhibition of berberine on Aβ40/42 and β-secretase and berberine-induced activation of p-ERK1/2.Conclusions The present study suggests that Berberine inhibits the production of Aβ40/42 and expression of β-secretase via activation of ERK1/2 pathway.