中国汉族人群Toll 样受体4 基因多态性与阿尔茨海默病 相关性研究

目的:探索中国汉族人群Toll样受体4基因Asp299Gly多态性与阿尔茨海默病(AD)相关性。方法:样本来源于2010.01-2012.01上海普陀区人民医院门诊及病房、瑞金医院、华山医院痴呆专病门诊就诊的200例AD患者(年龄在45-85岁之间)与同时期入组的患者配偶或体检中心200例健康对照(统计分析时有10例患者因故剔除),所有入组者为无血缘关系的中国汉族人。AD诊断由神经科专科医生参照美国国立神经病学、语言交流障碍和卒中老年性痴呆和相关疾病学会工作小组(NINCDS-ADRDA)关于AD的诊断标准确定。所有入组者抽取随机静脉血2 ml,血样标本使用双脱氧链终止法进行TLR4基因Asp299Gly突变检测。实验前期随机抽取部分血样进行TLR4蛋白定量测定。因为使用双脱氧链终止法未检测到TLR4基因Asp299Gly突变型,在实验后期我们又使用连接酶检测反应法对其中352例样本进行TLR4基因Asp299Gly突变复测及ApoE等位基因型检测。结果:与对照组相比,AD组外周血TLR4蛋白含量增高,其差异性具有统计学意义(P<0.05)。基因检测结果显示TLR4基因Asp299Gly突变两组均为野生型A,AD组ApoEε4基因型频率(包含纯合子及杂合子)高达35.90%,明显高于对照组(17.35%)。结论:AD组外周血TLR4蛋白表达量增高提示TLR4与AD发病有一定相关性,但没有证据表明TLR4基因Asp299Gly突变与AD发病有相关性。

Relationship between Toll-like Receptor 4 Single Nucleotide Polymorphisms and Alzheimer Disease in Chinese Han Population*

Objective: To explore the relationship between Toll-like Receptor 4 single nucleotide polymorphisms and Alzheimer disease in the Chinese Han population.Methods: There were 200 AD patients(aged 45 to 85 years) and 200 health controls(aged 45 to 85 years) enrolled from January 2010 to January 2012.The patients were consecutively recruited from the outpatient and inpatient department in Shanghai Putuo District People’s Hospital,Clinic for dementia in Huashan Hospital and Ruijin Hospital.The health volunteers were recruited from spouses of the patients and the general physical examination population.All the enrolled subjects had no blood relationship.AD diagnosis were verified by an experienced neurological specialist according to the NINCDS-ADRDA criteria.2 ml peripheral blood was taken from all the subjects for TLR4-Asp299Gly mutation test by the dideoxy chain termination method.Meanwhile,TLR4 protein quantitative determination was finished for some random samples in both two groups at the early stage of the experiment.Additionally,in the later stage of experiment,352 of the samples were retested for TLR4-Asp299Gly mutation through Polymerase Chain Reaction-Ligase Detection Reaction(PCR-LDR) method and the ApoE genotype was detected through Snapshot method since the TLR4-Asp299Gly mutation had been not detected through dideoxy chain termination method.Results: The content of TLR4 protein is higher in the AD group than that in the control group,and the difference is statistically significant(P <0.05).Asp299Gly mutation test result shows that the two groups of the samples are wild genotype A without mutation of G,and the frequency(including homozygous and heterozygous) of ApoE ε4 genotype is significantly higher in the AD group with 35.9 %,which is much higher than that of the control group(17.35 %).Conclusions: In contrast to the control group,the higher expression of TLR4 in peripheral blood of AD group indicates that TLR4 plays a role in AD process,but evidence on correlation between Asp299Gly mutation of TLR4 gene and the incidence of AD is not observed.