尿酸介导OPRD1基因低甲...化影响阿尔茨海默病（英文）_陈瑶.pdf

第36卷第4期,2023年7月 宁 波 大 学 学 报（理 工 版）中国科技核心期刊 Vol.36 No.4,July 2023 JOURNAL OF NINGBO UNIVERSITY(NSEE)中国高校优秀科技期刊 DOI:10.20098/ki.1001-5132.2023.0414 Uric acid mediated hypomethylation of OPRD1 gene affects Alzheimers disease CHEN Yao1,2,CHEN Yong1*,YING Ying1,FAN Jianbo3,GENG Baoqing1,ZHENG Chengying4,XU Ting4,FU Haoyuan5,ZHANG Yunge6,CHEN Ji6(1.Ningbo No.2 Hospital,Ningbo 315010,China;2.Taizhou Hospital of Zhejiang,Taizhou 317099,China;3.Zhenhai Longsai Hospital,Ningbo 315299,China;4.Ningbo Kangning Hospital,Ningbo 315201,China;5.The Third Peoples Hospital Health Care Group of Cixi(Chang He Branch),Ningbo 315325,China;6.Health Science Center,Ningbo University,Ningbo 315211,China)Abstract:The present study was aimed to investigate the correlation between OPRD1 gene methylation,uric acid and Alzheimers disease.It also examined whether uric acid could affect OPRD1 gene methylation in vitro.The venous blood of the patients with Alzheimers disease,the hyperuricemia group and the healthy control group was collected.The cells were successively treated with different concentrations of uric acid and the methylation level of OPRD1 gene was subsequently determined.The methylation levels of OPRD1 at pos2 and pos4 in the hyperuricemia group were lower than those of the healthy control group;while the methylation level of pos4 in the Alzheimers disease group was higher than that of the healthy control group.Uric acid levels were negatively correlated with OPRD1 gene methylation levels.In vitro,the methylation levels at pos2,pos3 and pos4 sites of OPRD1 gene in cells with high concentration of uric acid were lower even further.The present study suggested that uric acid may affect the onset of Alzheimers disease by mediating the hypomethylation modification at the pos4 site of OPRD1 gene.Key words:uric acid;key enzymes;OPRD1 gene;methylation;Alzheimers disease CLC number:R741 Document code:A Article ID:1001-5132（2023）04-0078-11 1 Introduction Alzheimers disease(AD),the most common neurodegenerative disease leading to senile dementia,is a major public health problem worthy of the attention of the whole society.Recent studies have found that hypermethylation modification of the OPRD1 gene is associated with the risk of AD1.The-opioid receptor(DOR)encoded by the OPRD1 gene is involved in protecting neurons from hypoxic/ischemic stress.A research displays that the distribution of DOR in the brain of patients with AD is significantly reduced2.In addition,quantitative autoradiography found that the binding of DOR in the amygdala complex and ventral 收稿日期:20230413.宁波大学学报（理工版）网址:http:/ 第 4 期 CHEN Yao,et al:Uric acid mediated hypomethylation of OPRD1 gene affects Alzheimers disease 79 putamen in the brain of patients with AD decreased3.This indicates that the OPRD1 gene has a strong correlation with DOR and AD,which may be due to the down-regulation of OPRD1 gene expression in the hypermethylated state and the inhibition of DOR coding,leading to an increased risk of AD.Uric acid is the end product of purine metabolism in the human body.At present,there is increasing evidence that uric acid may play a certain neuroprotective role in the occurrence and development of AD.A meta-analysis in 2016 found that serum uric acid levels in patients with AD were lower than those in the healthy control group,and high uric acid levels were significantly associated with a reduced risk of AD4.In addition,a study on uric acid,AD and cognitive dysfunction has shown that high levels of uric acid can slow the progression of cognitive decline,especially statistically significant in patients with mild cognitive impairment and AD5.In the early stage of the research group,methylation microarray screening was performed on the genome of patients with gout and hyperuricemia6.The results showed that in patients with gout,the pos3 site in the promoter region of the OPRD1 gene was hypome-thylated,and in patients with hyperuricemia,the pos2 and pos4 sites in the promoter region of OPRD1 gene were hypomethylated.And through a large sample experiment of human peripheral blood,it was confirmed that the OPRD1 gene is hypomethylated and the OPRD1 protein is highly expressed in gout patients.Therefore,we have reason to believe that uric acid may be related to the hypomethylation of the OPRD1 gene.To sum up,the purpose of this study is to confirm the correlation between OPRD1 gene methylation level and uric acid level as well as AD in humans,and explore the correlation between uric acid and methylation modification at OPRD1 gene locus in cell experiments.The clinical significance of this study is to explore the pathogenesis of AD with uric acid level as the source,which provides a new idea and a theoretical basis for the treatment of AD.2 Materials and methods 2.1 Experiment 1:Correlation between OPRD1 gene methylation and uric acid and AD 2.1.1 Research objects 45 patients with AD were enrolled in the study.The diagnosis was consistent with the 1994“American Diagnostic an