Xu Peng-Bo, Ding Li-Dan, Qiu Jing-Wen, Zhong Hua, Wu Huan, Zhou An, Wu Hong-Fei, Dai Min
Anhui Province Key Laboratory of Research & Development of Chinese Medicine, Department of Pharmacy, Anhui University of Chinese Medicine Hefei 230012, China.
Zhongguo Zhong Yao Za Zhi. 2021 Oct;46(20):5320-5329. doi: 10.19540/j.cnki.cjcmm.20210702.403.
In this study, ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS)-based liver metabolomics approach was used to explore the mechanism of "Trichosanthis Fructus-Allii Macrostemonis Bulbus" in improving atherosclerosis(AS) of mice with apolipoprotein E gene knockout(ApoE~(-/-)). AS mouse model was induced by high-fat diet. The pathological and biochemical indexes such as the histopathological changes, body weight, liver weight, blood lipid level and inflammatory factors in the liver of mice were determined. The metabolic profiling of mice liver samples was performed with UPLC-Q-TOF-MS. Multiple statistical analysis methods including partial least squares discriminant analysis(PLS-DA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were employed to screen and identify biomarkers. The levels of related enzymes including LCAT, sPLA2, EPT1 and ACER1 were detected. The results showed that "Trichosanthis Fructus-Allii Macrostemonis Bulbus" significantly reduced the areas of aortic plaque and fat vacuoles of liver in AS mice and decreased the accumulation of lipid droplets and liver coefficient. "Trichosanthis Fructus-Allii Macrostemonis Bulbus" also regulated the levels of blood lipid and inflammatory injury in the liver. The metabolites of the control group, the model group and the "Trichosanthis Fructus-Allii Macrostemonis Bulbus" group could be distinguished significantly. Fifteen potential biomarkers related to AS were discovered and preliminarily identified, seven of which could be regulated by "Trichosanthis Fructus-Allii Macrostemonis Bulbus" in a trend of returning to normal. Metabolic pathway analysis screened out two major metabolic pathways. "Trichosanthis Fructus-Allii Macrostemonis Bulbus" obviously regulated the levels of LCAT, sPLA2, EPT1 and ACER1. It was inferred that "Trichosanthis Fructus-Allii Macrostemonis Bulbus" could play a major role in AS treatment by regulating glycerophospholipid and sphingolipid metabolism disorders in the liver, with the mechanism probably relating to the intervention of the expression of LCAT, sPLA2, EPT1 and ACER1.
在本研究中，采用基于超高效液相色谱-四极杆飞行时间串联质谱联用技术（UPLC-Q-TOF-MS）的肝脏代谢组学方法，探究“瓜蒌薤白方”改善载脂蛋白E基因敲除（ApoE~(-/-)）小鼠动脉粥样硬化（AS）的机制。通过高脂饮食诱导建立AS小鼠模型，测定小鼠肝脏的组织病理学变化、体重、肝脏重量、血脂水平及炎症因子等病理生化指标。采用UPLC-Q-TOF-MS对小鼠肝脏样本进行代谢轮廓分析，运用偏最小二乘法判别分析（PLS-DA）和正交偏最小二乘法判别分析（OPLS-DA）等多种统计分析方法筛选和鉴定生物标志物，检测卵磷脂胆固醇酰基转移酶（LCAT）、分泌型磷脂酶A2（sPLA2）、乙醇胺磷酸甘油转移酶1（EPT1）和酸性神经酰胺酶1（ACER1）等相关酶的水平。结果显示，“瓜蒌薤白方”显著减小了AS小鼠主动脉斑块面积和肝脏脂肪空泡面积，降低了脂质滴积累和肝脏系数。“瓜蒌薤白方”还调节了血脂水平及肝脏的炎症损伤。对照组、模型组和“瓜蒌薤白方”组的代谢产物能够被显著区分。发现并初步鉴定了15个与AS相关的潜在生物标志物，其中7个可被“瓜蒌薤白方”调节至趋于正常的水平。代谢通路分析筛选出两条主要代谢通路。“瓜蒌薤白方”明显调节了LCAT、sPLA2、EPT1和ACER1的水平。推测“瓜蒌薤白方”可能通过调节肝脏中甘油磷脂和鞘脂代谢紊乱在AS治疗中发挥主要作用，其机制可能与干预LCAT、sPLA2、EPT1和ACER1的表达有关。
