目的探讨补肾益智方有效部位群对M146L细胞分泌的β淀粉样蛋白的影响,寻找潜在的具有抑制Aβ分泌的中药复方有效群。方法体外培养稳定转染人类阿尔茨海默病(AD)β淀粉样前体蛋白(beta-amyloid protein precursor,APP)基因及突变型早老素1(prsenilin-1,PS1)基因的中华仓鼠卵巢上皮细胞系(M146L),使之高效产生β淀粉样蛋白Aβ1-42,建立Aβ1-42过度表达的细胞模型。加入待筛选的补肾益智方有效提取部位群,用MTT法检测不同浓度的补肾益智方有效提取部位群(0,12.5,50μg/ml)对M146L细胞的毒性作用,应用Western blot免疫印迹方法分别检测突变型PS1M146L和野生型APP751双转染稳定表达CHO细胞系分泌的Aβ1-42的变化。结果不同浓度的补肾益智方有效部位群对M146L存活均无影响,不具有细胞毒作用。其中浓度为12.5μg/ml有效部位群F1和F2对M146L细胞分泌Aβ1-42有抑制作用。结论一定剂量的补肾益智方有效提取部位群对M146L细胞分泌的Aβ1-42有明显的抑制作用,其机制有待进一步研究。
OBJECTIVE Bushen Yizhi formula,constructed by Prof LIAO Shi-long′research group(Guangzhou University of chinese medicine),is the combination of clinical experience and modern pharmacological research,and mainly focuses on the etiology and pathogenesis of AD for treating both cause and symptoms.In this research,the pharmacodynamic study protocol of Bushen Yizhi formula was designed according to the characteristics and indications function of herbs in the formula,and was conformed to preclinical pharmacological research of traditional Chinese medicine for AD,The Technical Requirements of Pharmacology and Toxicology Research in New Drug Development Enacted by State Food and Drug administration also was guiding principle.METHODS A preliminary randomized double-blind clinical trail with 141 cases of AD showed that the efficiency of Bushen Yizhi formula was 61.9%and the increased mean of MMSE was 3.17,but the specific mechanism remains unclear.This study was aimed to reveal the preventive and therapeutic effect of Bushen Yizhi formula on AD rats and its related mechanism.In this research,forebrain-injected IBO-induced AD rats(cholinergic neuron lesion),senescence accelerated mouse,scopolamineinduced learning and memory deficiency rats(mild cognitive impairment)were all established.The learning and memory ability was tested with Morris water maze.Then formation of age pigment,extent of neuronal loss,activation of astrocytes,content of NTFs and degree of oxidized stress damage were determined by morphology,mmunohistochemical and molecular biology methods.RESULTS As the application of Bushen Yizhi formula,the learning and memory ability in all three groups were significantly improved,the formation of age pigment and the content of ACH in cortex and hippocampus were reduced,the activation of astrocytes and release of inflammatory factor(TNF-αand IL-1β)were inhibited,and the antioxidases(CAT,SOD,GSH-PX)were up-regulated and MDA was down-regulated.CONCLUSION Bushen Yizhi formula can prevent and treat AD rats,which might
Objective： To establish the diagnostic quantitative criteria for fire-heat syndrome （FHS） of Chinese medicine （CM） based on the receiver operating characteristic （ROC） curve and principal component analysis （PCA）. Methods： The symptoms and signs of FHS cases and healthy subjects from Guangzhou, Henan and Hunan of China were collected through questionnaire, and the diagnostic quantitative score tables were established for the three regions, respectively, with the method of maximum likelihood analysis. The homogeneity test was then performed on the diagnostic score tables for the three regions with ROC curve, and the diagnostic efficiency of diagnostic score tables for the three regions was compared with the prospective test and retrospective test. The method of PCA was adopted to obtain the analysis matrix for classifying the tapes of FHS. Results： Twenty-seven elements of FHS were confirmed through Chi-square test, and the diagnostic score tables for the three regions were established with the method of maximum likelihood analysis on the basis of the collected case data. According to the ROC curve test, the areas under ROC curve of Guangzhou diagnostic score table assessment with candidates in Guangzhou, Henan and Hunan were 0.998, 0.961 and 0.956, respectively. It showed that the diagnostic efficiency of Guangzhou diagnostic score tables was the highest one. With the prospective test, the area under ROC of Guangzhou diagnostic score table was 0.949, and more than any other diagnostic score table. By PCA, FHS was classified into excess fire and deficiency fire, and then classified into syndrome of flaring up of Heart （Xin） fire, syndrome of Lung （Fei）-Stomach （Wei） excess fire, syndrome of deficiency of Liver （Gan）-yin and Kidney （Shen）-yin, and syndrome of deficiency of Lung-yin from the view of viscera. In the retrospective test, the consistency with clinicians＇ diagnosis was 69.4%, and in the prospective test, it was 70.1%. Conclusions： The Guangzhou diagnostic sco
