中文摘要：

【目的】探讨地黄饮子对Aβ_（1-42）诱导的SH-SY5Y细胞中RAGE/ROS/凋亡通路的影响。【方法】（1）采用四甲基偶氮唑盐（MTT）法检测胎牛血清组、空白组及地黄饮子含药血清低、中、高剂量组的细胞活力,确定地黄饮子含药血清的最佳浓度和作用时间。（2）以0~20μmol/L Aβ_（1-42）寡聚体处理SH-SY5Y细胞24 h和48 h后,采用MTT法检测细胞活力,Annexin V/碘化丙啶（PI）双染法观察细胞凋亡,确定Aβ_（1-42）作用细胞的最佳浓度及时间,以建立阿尔茨海默病（AD）细胞模型。（3）采用MTT法检测空白组、模型组、西药对照组及地黄饮子含药血清低、中、高剂量组细胞活力,采用Annexin V/PI双染法观察各组细胞凋亡,采用二氢乙啶（DHE）染色法检测各组活性氧（ROS）含量,观察地黄饮子对Aβ_（1-42）诱导的SH-SY5Y的细胞损伤的修复作用。（4）采用Western blot法检测空白组、模型组、地黄饮子含药血清中剂量组RAGE蛋白;进一步将Aβ_（1-42）诱导的SH-SY5Y细胞进行RAGE转染后,采用DHE染色法检测各组ROS含量,采用Annexin V/PI双染法检测各组细胞凋亡率,观察地黄饮子对Aβ_（1-42）诱导的SH-SY5Y细胞凋亡及RAGE表达的影响。【结果】作用时间为24 h时,地黄饮子含药血清低、中剂量组细胞活力较空白组均显著增强（P〈0.05或P〈0.01）。建立AD体外模型的Aβ_（1-42）浓度为5μmol/L,作用时间为24 h。各给药组Aβ_（1-42）诱导细胞活力较模型组显著增强,细胞凋亡率和ROS含量显著下降（P〈0.05或P〈0.01）,而地黄饮子中剂量组细胞活力最强,细胞凋亡率最低。地黄饮子中剂量组Aβ1-42诱导细胞RAGE蛋白表达较模型组显著降低（P〈0.05）。地黄饮子中剂量组能降低RAGE转染的Aβ_（1-42）诱导SH-SY5Y细胞的ROS生成和细胞凋亡率（P〈0.01）。【结论】地黄饮子可能通过抑制ROS产生和细胞凋亡发挥抗氧化作用,进而抑制RAG

英文摘要：

Objective To investigate the effects of Dihuang Yinzi （DY） on the receptor for advanced glycation end-products（RAGE）/reactive oxygen species（ROS）/apoptosis pathway in SH-SY5Y cells induced by amyloid-beta1-42 （Aβ1-42） oligomer. Methods Firstly, we adopted methyl thiazolyl tetrazolium（MTT） method to detect the cell vitality in fetal bovine serum （FBS） group, blank serum group, and low-, middle- and high- dose DY-containing serum groups, so as to confirm the optimal concentration and treatment time of DY-containing serum. Secondly, we applied MTT method to detect cell vitality and applied Annexin V/propidium iodide （PI） staining method to observe the apoptosis of SH-SY5Y cells treated with 0~20 μmol/L Aβ1-42 for 24 and 48 h, so as toconfirm the optimal concentration and treatment time of Aβ1-42 for establishing Alzheimer＇s disease （AD） model in vitro. Thirdly, MTT method was used for the detection of cell vitality, and Annexin V/PI staining method was used for detection of the apoptosis of SH-SY5Y cells in blank serum group, model group, western medicine control group and low-, middle-and high-dose DY-containing serum groups, and Dihydroethidium （DHE） method was used for the assay of ROS contents, so as to observe the effect of DY on the recovery of injured SH-SY5Y cells induced by Aβ1-42. Finally, we applied Western blot method to detect the expression level of RAGE in SH-SY5Y cells of blank group, model group and DY-containing serum group; after Aβ1-42-induced SH-SY5Y cells were transfected with RAGE gene, we adopted DHE staining method and Annexin V/PI staining method to detect ROS content and cell apoptotic rate in all of the above groups, so as to observe the effect of DY on SH-SY5Y cell apoptosis and RAGE expression. Results The cell vitalities were increased in low- and middle-dose DY-containing serum groups at 24 h （P 〈 0.05 or P 〈 0.01 compared with that in the blank serum group）. The conditions for the establishment of AD model in vitro were as follows?