摘要
目的探讨地黄饮子对β-淀粉样蛋白1-42（Aβ1-42）诱导的人神经母细胞瘤细胞株（SH-SY5Y）晚期糖基化终产物受体（RAGE）/p38丝裂原活化蛋白激酶（p38 MAPK）/核转录因子kappa-B（NF-κB）信号通路的影响。方法将雄性SD大鼠按随机数字表法分为正常组及给药组（每组20只），分别配制正常血清培养液及地黄饮子含药血清培养液。（1）将SH-SY5Y细胞分为3组，其中对照组采用正常血清培养液培养，模型组采用正常血清培养液＋Aβ1-42寡聚体培养，地黄饮子组采用地黄饮子含药血清培养液＋Aβ1-42寡聚体培养。采用Western blotting检测各组SH-SY5Y细胞NF-κB p65、p38、磷酸化（p）-p38蛋白表达。（2）将SH-SY5Y细胞单独设为转染RAGE地黄饮子组，于Aβ1-42寡聚体作用不同时间（15 min、30 min、60 min、12 h、24 h、48 h、72 h）时，采用Western blotting检测各时间点SH-SY5Y细胞p-p38、p38蛋白表达。（3）将SH-SY5Y细胞分为6组，其中模拟转染RAGE对照组采用正常血清培养液＋空载体质粒转染培养，转染RAGE对照组采用正常血清培养液＋RAGE质粒转染培养，模拟转染RAGE模型组采用正常血清培养液＋Aβ1-42寡聚体＋空载体质粒转染培养，转染RAGE模型组采用正常血清培养液＋Aβ1-42寡聚体＋RAGE质粒转染培养，模拟转染RAGE地黄饮子组采用地黄饮子含药血清培养液＋Aβ1-42寡聚体＋空载体质粒转染培养，转染RAGE地黄饮子组培养方法同前。采用ELISA法和流式微球分析（CBA）法检测各组SH-SY5Y细胞炎症因子白介素（IL）-1β、IL-6、肿瘤坏死因子（TNF）-α含量。结果（1）与模型组比较，地黄饮子组SH-SY5Y细胞NF-κB p65蛋白、p-p38/p38表达水平明显降低，差异均有统计学意义（P〈0.05）。（2）p-p38蛋白表达水平在Aβ1-42作用30 min时开始明显升高，持续至24 h时表达最高，48 h时开始出现下调趋势。（3）模拟转染RAGE模�
Objective To investigate the effect of Dihuangyinzi （DHYZ）-medicated serum on receptor for advanced glycation end product （RAGE）/p38 miotgen-activated protein kinase （MAPK） /nuclear factor （NF）-κB pathway in SH-SY5Y cells induced by Aβ1-42. Methods Male SD rats were randomly divided into normal control group and experimental group （n=20）; natural sera medium and DHYZ sera medium were prepared. （1） SH-SY5Y cells were divided into control group, model group and DHYZ treatment group; natural sera medium, natural sera medium＋Aβ1-42 oligomer, and DHYZ sera medium＋Aβ1-42 oligomer were given to the cells, respectively. Western blotting was used to detect the protein expressions of NF-κB p65, p38 and phosphorylate （p）-p38. （2） SH-SY5Y cells were given DHYZ sera medium＋Aβ1-42 oligomer treatment, and at different time points of Aβ1-42 oligomer treatment （15 min, 30 min, 60 min, 12 h, 24 h, 48 h and 72 h）, Western blotting was used to detect the protein expressions of p38 and p-p38. （3） SH-SY5Y cells were divided into 6 groups： mock-transfected RAGE blank group, transfected RAGE blank group, mock-transfected RAGE model group, transfected RAGE model group, mock-transfected RAGE herb group and transfected RAGE herb group; herb groups were given DHYZ-medicated serum; inflammatory factors, interleukin （IL）-1β, IL-6, and tumor necrosis factor （TNF）-α, were measured by ELISA and cytometric bead array. Results （1） As compared with model group, DHYZ treatment group had significantly decreased NF-KB p65 and p-p38/p38 protein expression. （2） The p-p38 protein expression began to increase 30 rain after Aβ1-42 treatment, reached to its peak level 24 h after Aβ1-42 treatment, and began to decrease 48 h after Aβ1-42 treatment. （3） The IL-113, IL-6 and TNF-α levels were increased significantly in the transfected RAGE model group as compared with those in the mock-transfected RAGE model group （P〈0.05）; the IL-1β, IL-6 and TNF-α levels were increased si
作者
朴钟源
魏亚芬
宋琳
姚丽芬
姜卓
陆一婵
郑杨
邸智勇
Piao Zhongyuan Wei Yafen Song Lin Yao Lifen Jiang Zhuo Lu Yichan Zheng Yang Di Zhiyong(Unite One, Department of Neurology, Xiangfang Branch, Heilongjiang Provincial Hospital, Harbin 150036, China Department of Experimental Diagnosis, Xiangfang Branch, Heilongjiang Provincial Hospital, Harbin 150036, China Department of Basic Theories of Chinese Medicine, Heilon~iang University of Chinese Medicine, Harbin 150040, China Department of Neurology, First Affiliated Hospital of Harbin Medical University, Harbin 150001, China)
出处
《中华神经医学杂志》
CSCD
北大核心
2017年第10期1022-1027,共6页
Chinese Journal of Neuromedicine
基金
国家自然科学基金（81673860、81271208、81302904）
黑龙江省自然科学基金（ZD2015018）
