基于肝脏代谢组学研究清脂通脉颗粒对ApoE^(-/-)动脉粥样硬化的影响  

Effect of Qingzhi Tongmai Granule(清脂通脉颗粒)on ApoE^(-/-)Atherosclerosis Based on Liver Metabolomics

作　　者：刘海云[1,2] 于游[1] 张瀚文[1] 张欢[1] 刘倩[1] 于睿[1] LIU Haiyun;YU You;ZHANG Hanwen;ZHANG Huan;LIU Qian;YU Rui(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;The Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《中华中医药学刊》2022年第12期132-135,I0049-I0053,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：中国博士后科学基金第70批面上资助“地区专项支持计划”(2021MD703843);辽宁省“兴辽英才计划”(XLYC1802004);辽宁省教育厅科学技术研究项目(L201948)。

摘　　要：目的观察清脂通脉颗粒对ApoE^(-/-)动脉粥样硬化小鼠肝脏代谢组学及炎症因子的影响,以期进一步从肝脾同调角度探讨清脂通脉颗粒防治动脉粥样硬化(atherosclerosis,AS)作用机制。方法将C57BL/6J雄性小鼠设为空白对照组(ZC),将AopE^(-/-)雄性小鼠随机分为模型对照组(MX)、依折麦布组(MB)、清脂通脉中剂量组(TM1)、清脂通脉高剂量组(TM2),每组8只。采用高脂饲料饲养制备AS动物模型。MB组、TM1组、TM2组每日灌胃相应药物治疗,ZC组、MX组每日灌胃等体积生理盐水,连续12周,每周记录小鼠体质量和一般状态。试验结束后处死小鼠收集主动脉组织、肝脏组织及血用于后续分析。采用HE染色观察主动脉与肝脏组织形态学变化。应用酶联免疫吸附技术检测小鼠血清脂多糖(lipopolysaccharide,LPS)、白介素-1β(interleukin-1β,IL-1β)、白介素-18(interleukin-18,IL-18)表达水平。采用UPLC-Q-ExactiveMS技术对肝脏差异代谢物进行分析。结果清脂通脉颗粒可明显改善ApoE^(-/-)动脉粥样硬化小鼠主动脉、肝脏组织病理变化。与ZC组比较,MX组小鼠LPS、IL-1β、IL-18水平显著升高(P<0.01);与MX组比较,MB组、TM1组、TM2组小鼠LPS、IL-1β、IL-18水平显著降低(P<0.01)。ZC组、MX组以及TM2组的代谢物能够显著区分,发现并初步鉴定与AS相关的胱氨酸、胆酸、柠檬酸三乙酯等潜在生物标志物,代谢通路分析包括烟酸和烟酰胺代谢、牛磺酸和亚牛磺酸代谢、淀粉和蔗糖代谢等多种代谢途径。结论清脂通脉颗粒能够调控ApoE^(-/-)动脉粥样硬化小鼠肝脏代谢,降低肝脏脂滴积累及炎性反应,抑制AS疾病发生发展,具有积极防治作用。Objective To observe the effects of Qingzhi Tongmai Granule(清脂通脉颗粒)on liver metabolomics and inflammatory factors in ApoE^(-/-)atherosclerotic mice,in order to further explore the mechanism of Qingzhi Tongmai Granule in the prevention and treatment of atherosclerosis(AS).Methods C57BL/6J male mice were set as blank control group(ZC).ApoE^(-/-)male mice were randomly divided into model control group(MX),ezetimibe group(MB),Qingzhi Tongmai Granule medium dose group(TM1)and Qingzhi Tongmai Granule high dose group(TM2),with 8 mice in each group.The animal model of AS was prepared by feeding with high-fat diet.MB group,TM1 group and TM2 group were treated with the corresponding drugs every day,ZC group and MX group were treated with the equal volume of normal saline every day for 12 weeks,and the body weight and general state of mice were recorded every week.After the experiment,the mice were killed and the aortic tissue,liver tissue and blood were collected for subsequent analysis.The histomorphological changes of aorta and liver were observed by HE staining.Serum levels of lipopolysaccharide(LPS),interleukin-1β(IL-1β)and interleukin-18(IL-18)were detected by enzyme-linked immunosorbent assay.The liver differential metabolites were analyzed by UPLC-Q-ExactiveMS.Results Qingzhi Tongmai Granule could significantly improve the histopathological symptoms of aorta and liver in ApoE^(-/-)atherosclerotic mice.Compared with those of the ZC group,levels of LPS,IL-1β and IL-18 in the MX group increased significantly(P<0.01).Compared with those of the MX group,the levels IL-1βand IL-18 in MB group,TM1 group and TM2 group decreased significantly(P<0.01).The metabolites of ZC group,MX group and TM2 group can be significantly distinguished.Potential biomarkers such as cystine,cholic acid and triethylcitrate related to AS were found and preliminarily identified.The metabolic pathway analysis included nicotinate and nicotinamide metabolism,taurine and hypotaurine metabolism,starch and sucrose metabolism and other met

关 键 词：清脂通脉颗粒 动脉粥样硬化 肝脏 代谢组学 炎症因子 

分 类 号：R256.22[医药卫生—中医内科学]