| 摘要: |
| 【目的】研究车前子提取物干预扑热息痛（Acetaminophen，APAP）引起小鼠肝损伤的作用，并探究其潜在作用机制。【方法】运用半仿生酶法提取车前子活性成分，采用液相色谱-质谱联用（LC MS）检测其主要成分。体内研究车前子提取物对APAP肝损伤的保护作用，将60只雌性昆明小白鼠随机分为正常对照组（NC）、APAP肝损伤模型组(LD)、车前子对照组(PA)以及APAP干预的车前子提取物高、中、低剂量组（HPA+LC、MPA+LD、LPA+LD）。其中NC组和LD组小鼠灌胃生理盐水、PA组小鼠灌胃200 μg/mL车前子提取物，HPA+LD,MPA+LD和LPA+LD组小鼠分别灌胃车前子提取物200,100,50 μg/mL,每天2次，连续6 d；末次给药12 h后，采血并分离肝脏，检测血清谷丙转氨酶（ALT）和谷草转氨酶（AST）活性，评价肝脏损伤程度；检测肝脏中超氧化物歧化酶（SOD）活性及谷胱甘肽（GSH）、过氧化氢（H2O2）和丙二醛（MDA）含量，评价肝脏的氧化应激程度；检测CYP2E1 mRNA和蛋白表达，并通过细胞色素P450酶2E1（Cytochrome 2E1，CYP2E1）法体外筛选车前子抗APAP肝损伤可能的有效成分。【结果】LC-MS检测发现，车前子提取物有37种主要成分；血清生化指标检测发现，不同剂量的车前子提取物明显减少APAP诱导的氧化产物，增强抗氧化防御能力，减轻肝脏损伤且呈剂量依赖性；车前子提取物显著降低CYP2E1 mRNA和蛋白表达，说明车前子抑制CYP2E1的活性；车前子提取物中抗APAP肝损伤的有效成分为车前草苷D、车前草苷E、高车前素和高车前苷。【结论】车前子提取物的有效成分为车前草苷D、车前草苷E、高车前素和高车前苷。车前子提取物对APAP诱导的肝损伤具有保护作用，其潜在作用机制可能与抑制肝脏CYP2E1的表达和活性有关。 |
| 关键词: 车前子提取物 扑热息痛 肝损伤 |
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| 基金项目:甘肃农业大学盛彤笙基金项目（GSAU-STS-1729）；甘肃农业大学科学人才引进专项（GSAU-RCZX201702）；甘肃农业大学动物医学院教研产学支持项目（JYCX-KX017） |
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| Inhibitory effect of active components of Plantago asiatica extraction on acetaminophen-induced liver injury in mice |
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DAI Guonian,WANG Guirong,WANG Meng,et a l
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| Abstract: |
| 【Objective】This study aimed to explore the effect of Plantago asiatica (PA) on liver injury induced by acetaminophen (APAP) in mice and its potential mechanism.【Method】In this experiment,active components of PA were acquired by the semi-bionic enzyme method and analyzed by liquid chromatography mass spectrometry (LC-MS).A total of 60 female Kunming mice were randomized into 6 groups,including normal control group (NC),APAP liver injury model group (LD),PA control group (PA),high,medium and low doses of PA (200,100 and 50 μg/mL) treatment groups (HPA+LD,MPA+LD and LPA+LD).The mice were intragastrically administrated with PA twice a day.Blood samples were collected,and liver was isolated after 12 h of the last administration.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected to evaluate the degree of liver injury.The contents of liver superoxide dismutase (SOD),glutathione (GSH),hydrogen peroxide (H2O2) and malondialdehyde (MDA) were detected to evaluate the degree of oxidative stress of liver.The expression and activity of CYP2E1 were measured in gene and protein levels in vivo.The active components of PA were screened by CYP2E1 enzyme in vitro.【Result】The PA extraction had 37 main components by LC-MS analysis.The PA extraction significantly reduced oxidation products and enhanced antioxidant defense activity in a dose-dependent manner by detecting the serum biochemical indicators.PA extraction decreased the CYP2E1 expression and activity.The active components of PA screened by CYP2E1 enzyme in vitro were plantasioside D,plantasioside E,hispidulin and homoplantaginin.【Conclusion】The active components in PA extraction were plantasioside D,plantasioside E,hispidulin and homoplantaginin.This study demonstrated that PA extraction protected liver injury induced by APAP and its potential mechanism was related to the CYP2E1 inhibition. |
| Key words: Plantago asiatica extraction acetaminophen liver injury |
