基于1H-NMR代谢组学的豨莶草水洗脱部位致大鼠肺损伤作用机制分析

目的:应用代谢组学分析豨莶草水洗脱部位(SWES)致大鼠血清和肺脏中内源性代谢产物的动态变化,探讨其造成肺损伤的可能作用机制,寻找肺损伤的敏感标志物。方法:将SD大鼠随机分为3组,即正常组(生理盐水)和SWES高、低剂量组(给药剂量分别为0.500,0.125 g·kg-1·d-1),给药4周,停药2周,检测大鼠体质量,并对肺脏组织进行病理学检查。采用血清和肺脏样本进行核磁共振氢谱(1H-NMR)检测,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)对大鼠体内代谢产物的变化进行分析。结果:SWES高剂量组大鼠出现肺脏炎症,停药2周后呈可逆性改变。SWES高剂量组与正常组代谢谱差异区分显著。血清和肺脏中共鉴定出11个差异性代谢产物,其中血清4个(乙酸、氧化三甲胺、甘氨酸、肌肉肌醇),肺脏9个(乳酸、乙酸、磷脂酰胆碱、丙酮酸、二甲胺、谷氨酸、甘油磷酸胆碱、甘氨酸、黄嘌呤)。肺损伤涉及丙酮酸代谢、谷氨酸代谢等通路的紊乱。停药后肺脏中大多数差异代谢产物水平显著回调,与病理检测相一致。结论:SWES造成的损伤器官之一为肺脏,肺损伤呈可逆性改变,肺损伤机制与能量代谢和氧化应激反应有关。

Mechanism Analysis of Lung Injury Induced by Water Elution Section of Siegesbeckiae Herba on Rats by 1H-NMR Metabolomics

Objective Metabolomics was used to analyze the dynamic changes of endogenous metabolites in serum and lung tissue of rats treated with water elution section of Siegesbeckiae Herba (SWES), and to explore the possible mechanism of lung injury and to search for the sensitive markers in serum and lung tissue.Method SD rats were randomly divided into three groups, namely the normal group (normal saline), SWES high-dose group and SWES low-dose group (0.500, 0.125 g·kg^-1·d^-1). SWES was given for 4 weeks and stopped for 2 weeks. The weight and pathological examination were regarded as observation parameters. Serum and lung tissue samples were detected by nuclear magnetic resonance hydrogen spectrum (¹H-NMR). The metabolites in rats were analyzed by partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA).Result Lung inflammation was shown in SWES high-dose group and returned to normal after withdrawal for 2 weeks. The metabolic spectrum of SWES high-dose group was significantly different from the normal group. There were 11 metabolites were identified by ¹H-NMR metabolomics, four differential metabolites were identified in serum acetate, trimethylamine oxide (TMAO), glycine, myo-inositol] and nine differential metabolites were identified in lung tissue lactate, acetate, phosphatidylcholine (PC), pyruvate, dimethylamine, glutamate, glycerophosphocholine (GPC), glycine, xanthine]. Lung injury was related to the disorder of pyruvate metabolism, glutamate metabolism and other pathways. Most of the metabolites in lung tissue had obviously levels of callback after drug withdrawal, which coincided with the pathological examination.Conclusion The lung is one of the damaged organs caused by SWES, and the lung injury is reversible and may be related to energy metabolism and oxidative stress.