六味地黄丸对秀丽隐杆线虫转基因AD模型治疗作用及机制的代谢组学研究  

A metabolomic study on the therapeutic effect and mechanism of Liuwei Dihuang pills on Caenorhabditis elegans transgenic AD model

作　　者：左坤 佟玉良 孙婷[3] 关雨佳 王迪[1] 王岩[3] 张宁[1,2,3] 雷霞[1,3,4] ZUO Kun;TONG Yu-lang;SUN Ting;GUAN Yu-jia;WANG Di;WANG Yan;ZHANG Ning;LEI Xia(Jiamusi University,Jiamusi 154007,China;Heilongjiang University of Chinese Medicine Jiamusi College,Jiamusi 154007,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China;Wuxi Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Wuxi 214071,China)

机构地区：[1]佳木斯大学,佳木斯154007 [2]黑龙江中医药大学佳木斯学院,佳木斯154007 [3]黑龙江中医药大学,哈尔滨150040 [4]南京中医药大学无锡附属医院,无锡214071

出　　处：《药物分析杂志》2023年第8期1360-1368,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金面上项目(82174007);中央支持地方高校改革发展资金人才培养项目(二至丸防治AD作用机理研究);黑龙江省卫生健康委科研课题(2020-303);黑龙江中医药大学基金项目(2018RCD19,2018pt04);国家自然科学基金青年项目(82003975)。

摘　　要：目的:观察六味地黄丸对Aβ转基因秀丽隐杆线虫模型学习记忆能力的影响,并采用代谢组学法对其作用机制进行初步探讨。方法:以秀丽隐杆线虫野生型作为空白对照、转染人源APP基因的CL2355型为阿尔茨海默症(Alzheimer disease,AD)模型,六味地黄丸提取物给药(1.0 mg·mL^(-1))组,另设二甲双胍阳性药(50 mmol·L^(-1))组,通过化学趋向性试验验证六味地黄丸对AD模型线虫学习记忆能力的影响;采用代谢组学方法,筛选六味地黄丸治疗组与模型组的差异代谢物并进行代谢通路的富集,探讨六味地黄丸治疗AD的作用机制。结果:与空白组相比,模型组对氯化钠的趋向性指数显著降低,联想性学习行为受到显著抑制(P<0.01)。与模型组相比,阳性药组与六味地黄丸组线虫氯化钠的趋向性指数显著升高,联想性学习行为得到修复。本研究在AD模型线虫组织样品中检测出9种差异代谢产物,分别是半乳糖基羟基赖氨酸、溶血磷脂酰胆碱[18∶4(9Z,12Z,15Z)、14∶1(9Z)、16∶1(9Z)]、α-哌唑-5α-磷酸、三丁酸甘油酯、L-亮氨酸、溶血磷脂酰乙醇胺(0∶0/14∶0)、N-乙酰基-L-组氨酸,涉及甘油磷脂代谢,缬氨酸、亮氨酸和异亮氨酸代谢途径。结论:六味地黄丸改善AD模型线虫学习记忆障碍与调节甘油磷脂、缬氨酸、亮氨酸和异亮氨酸等通路密切相关。Objective:To observe the effect of Liuwei Dihuang pills on the learning and memory ability of the Aβtransgenic Caenorhabditis elegans model,and to explore its mechanism by metabolomics.Methods:The wild-type C.elegans was used as the blank control,and the CL2355 type transfected with human APP gene was used as the Alzheimer disease model.The extract of Liuwei Dihuang pills was administered(1.0 mg·mL^(-1))as a group,an⁃other metformin positive drug(50 mmol·L^(-1))group was set up as a positive group.The effect of Liuwei Dihuang pills on the learning and memory ability of AD model nematodes was verified by chemotaxis test;metabolomics method was used to screen the differential metabolites between Liuwei Dihuang pills treatment group and model group and enrich the metabolic pathways to explore Liuwei Dihuang pills Mechanism of action in the treatment of AD.Results:Compared with the blank group,the model group significantly reduced the tropism index of NaCl,and significantly inhibited the associative learning behavior(P<0.01).Compared with the model group,the tropism in⁃dex of Nematode NaCl in the positive drug group and the Liuwei Dihuang pills group was significantly increased,and the associative learning behavior was repaired.In this study,nine differential metabolites were detected in AD mod⁃el nematode tissue samples,namely galactosylhydroxylysine,lysophosphatidylcholine[18∶4(9Z,12Z,15Z),14∶1(9Z),16∶1(9Z)],α-piperazole-5α-phosphate,tributyrate,L-leucine,lysophosphatidylethanolamine(0∶0/14∶0),N-acetyl-L-histidine,involved in glycerophospholipid metabolism,valine,leucine and isoleucine metabolism.Conclusion:The improvement of learning and memory impairment in AD model nematodes by Liuwei Dihuang pills is closely related to the regulation of glycerophospholipid,valine,leucine and isoleucine pathways.

关 键 词：阿尔茨海默病 六味地黄丸 代谢组学 秀丽隐杆线虫 代谢通路 

分 类 号：R917[医药卫生—药物分析学]