二仙汤对去卵巢大鼠干预作用的LC-MS血清代谢组学分析

杨莹; 胡晶; 刘海霞; 刘燕君; 宋长恒; 马玉杰; 张治国; 陈彦静

doi:10.13422/j.cnki.syfjx.20231161

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二仙汤对去卵巢大鼠干预作用的LC-MS血清代谢组学分析

药物代谢 | 更新时间：2023-09-21

- 二仙汤对去卵巢大鼠干预作用的LC-MS血清代谢组学分析
  增强出版
- Analysis of Intervention of Erxiantang in Ovariectomized Rats by LC-MS Serum Metabolomics
- 中国实验方剂学杂志 2023年29卷第20期页码：118-125
- 作者机构：
  
  中国中医科学院中医基础理论研究所，北京100700
- 作者简介：
  
  杨莹，在读硕士，从事中医药防治心血管病的基础研究，Tel：010-64089029，E-mail：yangyingwac@163.com
  张治国，博士，研究员，从事中医药防治代谢性疾病的基础研究，E-mail：zzgtcm@163.com
  陈彦静，博士，教授，从事中医药防治心血管病的基础研究，E-mail：chenyj010@163.com
- 基金信息：
  
  中央级公益性科研院所基本科研业务费专项(YZ-202041;YZ-202150);中国中医科学院科技创新工程项目(CI2021A00107)
- DOI：10.13422/j.cnki.syfjx.20231161
  中图分类号： R22;R28;R969.1;O657
- 纸质出版日期：2023-10-20，
  
  网络出版日期：2023-06-07，
  
  收稿日期：2023-04-06，
- 稿件说明：
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杨莹,胡晶,刘海霞等.二仙汤对去卵巢大鼠干预作用的LC-MS血清代谢组学分析[J].中国实验方剂学杂志,2023,29(20):118-125.

YANG Ying,HU Jing,LIU Haixia,et al.Analysis of Intervention of Erxiantang in Ovariectomized Rats by LC-MS Serum Metabolomics[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(20):118-125.
杨莹,胡晶,刘海霞等.二仙汤对去卵巢大鼠干预作用的LC-MS血清代谢组学分析[J].中国实验方剂学杂志,2023,29(20):118-125. DOI： 10.13422/j.cnki.syfjx.20231161.

YANG Ying,HU Jing,LIU Haixia,et al.Analysis of Intervention of Erxiantang in Ovariectomized Rats by LC-MS Serum Metabolomics[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(20):118-125. DOI： 10.13422/j.cnki.syfjx.20231161.

摘要

目的

基于液相色谱-质谱法（LC-MS）探讨去卵巢大鼠血清内源性代谢物的改变及二仙汤的干预作用。

方法

将24只健康雌性SD大鼠随机分为假手术组、模型组和二仙汤组，每组8只，模型组和二仙汤组切除双侧卵巢组织，假手术组切除双侧腹腔内小块脂肪组织，术后2周开始灌胃给药。二仙汤组给予7.5 g·kg

-1

二仙汤进行灌胃，假手术组和模型组灌胃等体积蒸馏水。给药结束后，腹主动脉取血，采用LC-MS对大鼠血清进行非靶向代谢组学研究，结合正交偏最小二乘法判别分析（OPLS-DA）筛选差异代谢物，基于基因组百科全书（KEGG）数据库进行代谢通路分析并采用酶联免疫吸附测定法（ELISA）验证代谢通路关键酶的水平。

结果

代谢组学结果显示，模型组与假手术组间82个差异代谢物包括甘油磷脂、脂肪酰基、类固醇及其衍生物等14类成分，其中差异最显著的为甘油磷脂类。二仙汤可回调82个差异代谢物中的65个，其中11个具有统计学意义，主要为甘油磷脂中的磷脂酰胆碱（PC）和溶血磷脂酰胆碱（LysoPC），其次为类固醇和类固醇衍生物中的皮质酮和11-脱氧皮质醇；代谢通路分析表明模型组甘油磷脂代谢、类固醇激素生物合成等通路发生变化，并在二仙汤给药后得以回调。ELISA结果显示，与假手术组比较，模型组大鼠血清中甘油磷脂代谢物PC、LysoPC生成的关键代谢酶磷酸胆碱胞苷酰转移酶（CCT）、分泌型磷脂酶A2（sPLA2）及溶血磷脂酰胆碱酰基转移酶（LPCAT）水平均明显降低（

0.05，

0.01），胆碱磷酸转移酶1（CPT1）水平有所下降但差异无统计学意义；与模型组比较，二仙汤组CCT、sPLA2、CPT1水平均有显著升高（

0.01）。此外，与假手术组比较，模型组总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）水平显著升高（

0.01），高密度脂蛋白胆固醇（HDL-C）水平明显降低（

0.05）；与模型组比较，二仙汤组TC、TG、LDL-C水平显著降低，HDL-C水平显著升高（

0.01）。

结论

去卵巢大鼠内源性代谢物及相关代谢通路发生改变，二仙汤可回调部分差异代谢物及相关通路，如调节代谢酶CCT、sPLA2、CPT1，从而升高PC、LysoPC的水平以调控甘油磷脂代谢，进而改善去卵巢大鼠脂代谢紊乱等病理变化。

Abstract

Objective

To investigate the changes of endogenous metabolites in serum of ovariectomized rats and the effect of Erxiantang on them based on liquid chromatography-mass spectrometry（LC-MS）.

Method

Twenty-four healthy female SD rats were randomly divided into sham-operated group， model group and Erxiantang group（7.5 g·kg

-1

）， with 8 rats in each group. Bilateral ovarian tissues were excised in the model and Erxiantang groups， and small pieces of adipose tissues were excised in the abdominal cavity of the sham-operated group bilaterally， and gastric administration was started 2 weeks after surgery， and equal volumes of distilled water were gavaged in the sham-operated and model groups. After 12 weeks of administration， blood was collected from abdominal aorta， and non-targeted metabonomics was performed on rat serum by LC-MS， and orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to screen differential metabolites. Metabolic pathway analysis was performed based on Kyoto Encyclopedia of Genes and Genomes（KEGG）， and the levels of key enzymes of metabolic pathways were verified by enzyme-linked immunosorbent assay（ELISA）.

Result

The results of metabonomics showed that 82 differential metabolites between the model group and the sham-operated group were glycerophospholipids， fatty acyls， steroids and steroid derivatives， of which the most significant difference was glycerophospholipids. At the same time， Erxiantang could call back 65 out of 82 differential metabolites， of which 11 were statistically significant， mainly phosphatidylcholine（PC） and lysophosphatidylcholine（LysoPC） in glycerophospholipids， followed by corticosterone and 11-deoxycortisol in steroids and steroid derivatives. Metabolic pathway analysis showed that the pathways of glycerophospholipid metabolism and steroid hormone biosynthesis in model group were changed， and were recovered after the administration of Erxiantang. ELISA results showed that compared with the sham-operated group， serum levels of cholinephosphate cytidylytransferase（CCT）， secretory phospholipase A2（sPLA2） and lysophosphatidylcholine acyltransferase（LPCAT）， which were the key metabolic enzymes of glycerophospholipid metabolite PC and LysoPC， were significantly decreased in the model group（

0.05，

0.01）， and choline phosphotransferase 1（CPT1） levels decreased but the difference was not statistically significant， compared with the model group， the levels of CCT， sPLA2 and CPT1 were significantly increased in Erxiantang group（

0.01）. In addition， compared with the sham-operated group， the levels of cholesterol（TC）， triglyceride（TG） and low density lipoprotein cholesterol（LDL-C） were significantly increased in the model group（

0.01）， the high density lipoprotein cholesterol（HDL-C） level was decreased（

0.05）， compared with the model group， the levels of TC， TG and LDL-C were significantly decreased and the level of HDL-C was significantly increased in Erxiantang group（

0.01）.

Conclusion

Endogenous metabolites and related metabolic pathways in ovariectomized rats were altered， and Erxiantang can reverse some of the different metabolites and related pathways， such as regulating glycerophospholipid metabolism by regulating metabolic enzymes CCT， sPLA2 and CPT1 to increase the levels of PC and LysoPC， and then improve the pathological changes such as lipid metabolism disorder in ovariectomized rats.

关键词

二仙汤液相色谱-质谱法（LC-MS）代谢组学脂代谢甘油磷脂去卵巢大鼠内源性代谢物

Keywords

Erxiantangliquid chromatography-mass spectrometry（LC-MS）metabolomicslipid metabolismglycerophospholipidsovariectomized ratsendogenous metabolites

references

BAUSET C，MARTÍNEZ-ASPAS A，SMITH-BALLESTER S，et al.Nuts and metabolic syndrome：Reducing the burden of metabolic syndrome in menopause［J］.Nutrients，2022，14（8）：1677-1695.

STACHOWIAK G，PERTYŃSKI T，PERTYŃSKA-MARCZEWSKA M.Metabolic disorders in menopause［J］.Prz Menopauzalny，2015，14（1）：59-64.

赵宏宇，王玉，孙雪缘，等.基于PI3K/Akt信号通路探讨哈蟆油对卵泡发育作用的影响［J］.中国实验方剂学杂志，2021，27（21）：121-130.

林士毅.代谢综合征中医证候规律及与微观指标相关研究［D］.广州：广州中医药大学，2009.

ZHANG Z，XIANG L，ZHAO L，et al.The protective effect of Erxiantang against myocardial injury in menopausal rat model［J］. BMC Complement Altern Med，2018，18（1）：245-255.

姜燕华，王昱涵，刘燕君，等.二仙汤对去卵巢大鼠心肌微血管及血液流变学的影响［J］.中国实验方剂学杂志，2020，26（24）：59-67.

王昱涵，姜燕华，向丽华，等.二仙汤对去卵巢大鼠心功能和血压动态演变的干预作用［J］.中国实验方剂学杂志，2020，26（3）： 53-61.

WANG Y，JIANG Y，HU J，et al.Dynamic evolution of cardiac function and glucose and lipid metabolism in ovariectomized rats and the intervention effect of Erxiantang［J］. Evid Based Complement Alternat Med，2022，doi：10.1155/2022/8090868http://dx.doi.org/10.1155/2022/8090868.

胡晶，姜燕华，王昱涵，等.去卵巢大鼠肠道菌群结构和功能的变化及二仙汤的干预作用［J］.中国实验方剂学杂志，2022，28（12）：61-68.

MEDINA-CONTRERAS J，VILLALOBOS-MOLINA R，ZARAIN-HERZBERG A，et al.Ovariectomized rodents as a menopausal metabolic syndrome model. A minireview［J］. Mol Cell Biochem，2020，475（1/2）：261-276.

谢慧慧，王莹，杨硕，等.二仙汤对围绝经期大鼠心脏电生理的影响［J］.中国实验方剂学杂志，2019，25（18）：43-48.

QUINN M A，XU X，RONFANI M，et al.Estrogen deficiency promotes hepatic steatosis via a glucocorticoid receptor-dependent mechanism in mice［J］.Cell Rep，2018，22（10）：2690-2701.

JABALLAH A，SOLTANI I，BAHIA W，et al.The relationship between menopause and metabolic syndrome：Experimental and bioinformatics Analysis［J］. Biochem Genet，2021，59（6）：1558-1581.

生宁，王彩虹，贾志鑫，等.代谢组学探索复方二仙汤调节双侧去卵巢大鼠脂代谢紊乱特征［J］.药学学报，2021，56（9）：2403-2409.

HUANG C Y，LIN Y Y，HSU C C，et al.Antiapoptotic effect of exercise training on ovariectomized rat hearts［J］.J Appl Physiol，2016，121（2）：457-465.

ZHAO H，LI X，ZHANG D，et al.Integrative bone metabolomics-lipidomics strategy for pathological mechanism of postmenopausal osteoporosis mouse model［J］.Sci Rep，2018，doi：10.1038/s41598-018-34574-6http://dx.doi.org/10.1038/s41598-018-34574-6.

TANG X Y，DAI Z Q，SHI D F，et al.An UHPLC-MS/MS method for simultaneous determination of ten sex steroid hormones in ovariectomy-induced osteoporosis rat and its application in discovery of sex steroid hormones regulatory components of Xian-Ling-Gu-Bao capsule［J］.J Pharm Biomed Anal，2021，doi：10.1016/j.jpba.2020.113888http://dx.doi.org/10.1016/j.jpba.2020.113888.

ABSHIRINI M，CABRERA D，FRASER K，et al.Mass spectrometry-based metabolomic and lipidomic analysis of the effect of high fat/high sugar diet and greenshell（TM）mussel feeding on plasma of ovariectomized rats［J］.Metabolites，2021，11（11）：754-773.

ZHU Q，WU Y，MAI J，et al.Comprehensive metabolic profiling of inflammation indicated key roles of glycerophospholipid and arginine metabolism in coronary artery disease［J］.Front Immunol，2022，doi：10.3389/fimmu.2022.829425http://dx.doi.org/10.3389/fimmu.2022.829425

XU X，LUO Z，HE Y，et al.Application of untargeted lipidomics based on UHPLC-high resolution tandem MS analysis to profile the lipid metabolic disturbances in the heart of diabetic cardiomyopathy mice［J］.J Pharm Biomed Anal，2020，doi：10.1016/j.jpba.2020.113525http://dx.doi.org/10.1016/j.jpba.2020.113525.

LIBBY P，KOBOLD S.Inflammation：A common contributor to cancer，aging，and cardiovascular diseases-expanding the concept of cardio-oncology［J］.Cardiovasc Res，2019，115（5）：824-829.

GERGES S H，EL-KADI A.Sex differences in eicosanoid formation and metabolism：A possible mediator of sex discrepancies in cardiovascular diseases［J］.Pharmacol Ther，2022，234：108046.

LI X，WU K，DONG Q，et al.Overall adjustment acupuncture improves osteoporosis and exerts an endocrine-modulating effect in ovariectomized rats［J］.Front Endocrinol，2022，13：1074516.

PETERSSON M，EKLUND M，UVNäS-MOBERG K.Oxytocin decreases corticosterone and nociception and increases motor activity in OVX rats［J］.Maturitas，2005，51（4）：426-433.

SONG Y，NIE L，WEI D，et al.Serum cortisol-to-cortisone ratio and 11-deoxycortisol are associated with hypertension and blood pressure in Chinese rural population：A cross sectional study［J］.J Hypertens，2022，40（3），432-440.

薛黎明.基于蛋白质组学淫羊藿苷防治骨质疏松作用机理及药对“淫羊藿仙茅”配伍机制研究［D］.上海：第二军医大学，2012.

赵金椽，南洋，陈平平，等.知母多糖对甲亢热证大鼠肝脏代谢的影响［J］.中医药信息，2022，39（4）：31-35.

佘楷杰，杨婧雯，孟丹华，等.基于网络药理学和实验验证的二仙汤调治焦虑障碍的分子机制［J］.中国实验方剂学杂志，2022，28（19）：185-193.

CHEN Y，LV J，FU L，et al.Metabolome-wide association study of four groups of persistent organic pollutants and abnormal blood lipids［J］.Environ Int，2023，173：107817.

WANG B，TONTONOZ P. Phospholipid remodeling in physiology and disease［J］.Annu Rev Physiol，2019，81：165-188.

SHIROUCHI B，NAGAO K，INOUE N，et al.Effect of dietary omega 3 phosphatidylcholine on obesity-related disorders in obese Otsuka Long-Evans Tokushima fatty rats［J］.J Agric Food Chem，2007，55（17）：7170-7176.

MASTELLONE I，POLICHETTI E，GRÈS S，et al.Dietary soybean phosphatidylcholines lower lipidemia：Mechanisms at the levels of intestine，endothelial cell，and hepato-biliary axis［J］.J Nutr Biochem，2000，11（9）：461-466.

COLE L K，VANCE J E，VANCE D E.Phosphatidylcholine biosynthesis and lipoprotein metabolism［J］.Biochim Biophys Acta，2012，1821（5）：754-761.

TSELMIN S，SCHMITZ G，JULIUS U，et al.Acute effects of lipid apheresis on human serum lipidome［J］.Atheroscler Suppl，2009，10（5）：27-33.

VAN DER VEEN J N，KENNELLY J P，WAN S，et al.The critical role of phosphatidylcholine and phosphatidylethanolamine metabolism in health and disease［J］.Biochim Biophys Acta Biomembr，2017，1859（9 Pt B）：1558-1572.

BARBER M N，RISIS S，YANG C，et al.Plasma lysophosphatidylcholine levels are reduced in obesity and type 2 diabetes［J］.PLoS One，2012，7（7）：e41456.

HA C Y，KIM J Y，PAIK J K，et al.The association of specific metabolites of lipid metabolism with markers of oxidative stress，inflammation and arterial stiffness in men with newly diagnosed type 2 diabetes［J］.Clin Endocrinol （Oxf），2012，76（5）：674-682.

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