基于肝脏TMT标记定量蛋白质组学技术研究越鞠丸防治“双心疾病”的作用机制  

Mechanism of Yuejuwan in Prevention and Treatment of Psychological and Heart Diseases Based on Liver TMT Labeled Quantitative Proteomics

作　　者：张瀚文[1] 于嘉祥 石岩[1] 张文顺[1] 韩雪莹[2] 张欢[1] 曲超 申鑫惠 马贤德[1] 于睿[1] 于游 ZHANG Hanwen;YU Jiaxiang;SHI Yan;ZHANG Wenshun;HAN Xueying;ZHANG Huan;QU Chao;SHEN Xinhui;MA Xiande;YU Rui;YU You(Liaoning University of Traditional Chinese Medicine(TCM),Shenyang l10847,China;College of Pharmacy,Liaoning University of TCM,Dalian 116600,China;Second Affiliated Second Affiliated Hospital in Liaoning University of TCM,Shenyang 110034,China)

机构地区：[1]辽宁中医药大学,沈阳110847 [2]辽宁中医药大学药学院,辽宁大连116600 [3]辽宁中医药大学附属第二医院,沈阳110034

出　　处：《中国实验方剂学杂志》2023年第1期26-36,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点基础研究发展计划(973计划)项目(2013CB532004);辽宁省“兴辽英才计划”青年拔尖人才项目(XLYC2007121);中国博士后科学基金面上资助项目(2021M693853,2021M693852);辽宁省科技厅博士科研启动课题(2021-BS-183);中医脏象理论及应用教育部重点实验室2021年度开放基金项目(zyzx2109);国家中医药管理局中医药古籍文献和特色技术传承专项(GZY-KJS-2020-026)。

摘　　要：目的:观察越鞠丸治疗“双心疾病”的效果,并研究其中的作用机制。方法:选用6周龄健康雄性SPF级载脂蛋白E敲除(AopE^(-/-))小鼠30只、同源C57BL/6J小鼠10只进行实验。将30只AopE^(-/-)小鼠分为模型组、越鞠丸低剂量组(7.58 g·kg^(-1)·d^(-1))、越鞠丸高剂量组(30.32 g·kg^(-1)·d^(-1)),每组10只;C57BL/6J小鼠10只作为空白组,各组灌胃12周。饲养期间通过慢性不可预知应激刺激(CUMS)联合高脂饲料诱导的方法构建“双心疾病”模型。灌胃给药后,通过各组小鼠行为学实验结果[旷场实验(OFT)、糖水偏嗜实验(SPT)]、全自动生化分析仪检测小鼠血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、5-羟色胺(5-HT)、去甲肾上腺素(NE)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、总胆固醇(TC)、甘油三酯(TG)的含量及小鼠主动脉、肝脏油红O、苏木素-伊红(HE)染色,心肌组织Masson染色进行模型评价。最后通过肝脏TMT标记定量蛋白质组学技术研究其中的作用机制。结果:模型小鼠出现明显的抑郁、焦虑、兴趣丧失、绝望等表现,主动脉、肝脏病理观察也出现明显脂质沉积,心肌组织出现明显心肌纤维化增多的表现,越鞠丸灌胃给药后能改善“双心疾病”模型小鼠的上述症状与指标。与空白组比较,第12周模型组小鼠站立次数、中心区域累计时间、总移动距离、移动速度、糖水偏嗜率均显著下降(P<0.01),与模型组比较,越鞠丸低、高剂量组小鼠上述指标均显著降低(P<0.01)。取材后,与空白组比较,模型组AST、ALT、TG水平显著增多(P<0.01),5-HT、NE、HDL-C水平均显著下降(P<0.01)。肝脏TMT标记定量蛋白质组学结果提示“双心疾病”模型小鼠主要引起ApoE、UDP葡醛酸基转移酶家族1成员A5(UGT1A5)、FASN等蛋白表达水平的变化,涉及乙酰辅酶A代谢、对细菌的反应、细胞氨基酸分解代谢等过程,与肝脏代�Objective:To observe the effects of Yuejuwan in the treatment of psychological and heart diseases(PHD)and explore its mechanism.Method:Thirty 6-week-old healthy male SPF AopE^(-/-)mice and 10homologous C57BL/6J mice were selected for the experiment.The 30 AopE^(-/-)mice were divided into a model group,low-dose(7.58 g·kg^(-1)·d^(-1))and high-dose(30.32 g·kg^(-1)·d^(-1))Yuejuwan groups,with 10 mice in each group,and 10 C57BL/6J mice were assigned to the blank control group.Intragastrical administration lasted12 weeks.During feeding,the PHD model was induced by chronic unpredictable mild stress(CUMS)combined with high-fat diet in mice.After intragastric administration,the behavioral results[open field test(OFT)and sucrose preference test(SPT)]of mice in each group,the content of aspartic transaminase(AST),alanine aminotransferase(ALT),5-hydroxytryptamine(5-HT),noradrenaline(NE),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC),and triglyceride(TG)in serum of mice detected by the automatic biochemical analyzer,the oil red O staining and HE staining of aorta and liver and Masson staining of myocardial tissues were used for model evaluation.Finally,liver TMTlabeled quantitative proteomics was used to explore the mechanism of action.Result:The model mice showed obvious manifestations of depression,anxiety,loss of interest,and despair,manifest lipid deposition in the aorta and liver by pathological observation,and increased myocardial fibrosis in myocardial tissues.After intragastric administration of Yuejuwan,the above symptoms and indexes of the PHD model mice were improved.Compared with the blank control group,the model group showed decreased standing times,cumulative time in the central area,total moving distance,moving speed,and sucrose preference at week 12(P<0.01).Compared with the model group,the Yuejuwan groups showed decreased indexes mentioned above(P<0.01).After sample collection,AST,ALT,and TG levels in the model group were higher(P<0.01)and the

关 键 词：越鞠丸 双心疾病 防治 肝脏 TMT标记定量蛋白质组学 

分 类 号：R259[医药卫生—中西医结合] R256.2[医药卫生—中医内科学] R395[医药卫生—中医学] R714.252[哲学宗教—心理学]