龙胆酒炙前后对四氯化碳诱导小鼠肝损伤保护作用的对比研究  

Comparative study on protective effect of Gentianae Radix et Rhizoma before and after processing with wine against carbon tetrachloride induced liver damage in mice

作　　者：王景媛[1] 张舒雯 张桥[1] 赵重博[1] 翟思程 史亚军[1] WANG Jingyuan;ZHANG Shuwen;ZHANG Qiao;ZHAO Chongbo;ZHAI Sicheng;SHI Yajun(Shaanxi Traditional Chinese Medicine Processing Technology Inheritance Base,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Xianyang Key Laboratory for processing genuine medicinal materials,Haojing College of Shanxi University of Science&Technology,Xianyang 712046,China)

机构地区：[1]陕西中医药大学陕西省中药炮制技术传承基地,陕西咸阳日712046 [2]陕西科技大学镐京学院咸阳地区道地药材炮制重点实验室,陕西咸阳712046

出　　处：《中草药》2024年第18期6226-6237,共12页Chinese Traditional and Herbal Drugs

基　　金：陕西省重点研发计划项目(2024SF-YBXM-468);咸阳市科学技术研究与发展计划项目(2020k02-94);咸阳市2020年重点实验室项目(2020k04-02);国家级大学生创新创业训练计划项目(202210716021)。

摘　　要：目的探讨龙胆Gentianae Radix et Rhizoma酒炙前后对四氯化碳(carbon tetrachloride,CCl4)诱导的急性肝损伤小鼠模型的保肝作用,并通过代谢组学技术揭示其潜在作用机制。方法采用高效液相色谱及紫外分光光度计测定龙胆酒炙前后龙胆苦苷及多糖含量变化。将KM小鼠随机分为对照组、模型组、水飞蓟素(150 mg/kg)组及生龙胆低、中、高剂量(2.5、5.0、10.0 g/kg)组和酒龙胆低、中、高剂量(2.5、5.0、10.0 g/kg)组,每组6只。小鼠连续10 d ig相应药物,除对照组外其余小鼠ip含0.12%CCl4的花生油制备急性肝损伤模型。测定小鼠肝脏指数;采用苏木素-伊红(hematoxylin-eosin,HE)染色观察肝组织病理变化;采用试剂盒测定血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)活性及白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平和肝组织中谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性、丙二醛(malondialdehyde,MDA)水平;采用免疫组化法检测肝组织中磷酸化核因子-κB(phosphorylated nuclear factor-κB,p-NF-κB)表达;采用超高效液相色谱串联飞行时间质谱分析技术(UPLC-Q-TOF-MS)对各组小鼠血清代谢物进行分析。结果生、酒龙胆中龙胆苦苷质量分数分别为3.84%、3.12%,总多糖质量分数分别为3.27%、5.03%。与模型组比较,生、酒龙胆高剂量组小鼠肝损伤明显改善,血清中ALT、AST、ALP活性和TNF-α、IL-1β、IL-6水平显著降低(P<0.01),肝组织MDA水平和p-NF-κB表达显著降低(P<0.05、0.01),肝组织GSH-Px活性显著升高(P<0.01);与生龙胆高剂量组比较,酒龙胆高剂量组ALT、AST、ALP活性和TNF-α、IL-1β、IL-6水平显著降低(P<0.01),肝组织MDA水平和p-NF-κB表达显著降低(P<0.01),肝组织GSH-Px活性显著升高(P<0.01)。代谢组学研究表明龙胆酒�Objective To explore the hepatoprotective effect of Longdan(Gentianae Radix et Rhizoma)before and after processing with wine in mice with acute liver injury induced by carbon tetrachloride(CCl4),and to reveal the potential mechanism through metabolomics techniques.Methods The changes in contents of gentiopicroside and polysaccharides were determined by high-performance liquid chromatography and ultraviolet spectrophotometer.KM mice were randomly divided into control group,model group,silymarin(150 mg/kg)group,Gentianae Radix et Rhizoma low-,medium-,and high-dose(2.5,5.0,10.0 g/kg)groups and Gentianae Radix et Rhizoma processed with wine low-,medium-,and high-dose(2.5,5.0,10.0 g/kg)groups,with six mice in each group.Mice were administered the corresponding drug for 10 consecutive days,and except for the control group,acute liver injury mice were prepared by administering peanut oil containing 0.12%CCl4.The liver index of mice was measured.Hematoxylin-eosin(HE)staining was used to observe pathological changes in liver tissue.Kits were used to measure the activities of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),the levels of interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)in serum and glutathione peroxidase(GSH-Px)activity and malondialdehyde(MDA)level in liver tissue.Immunohistochemistry was used to detect the expression of phosphorylated nuclear factor-κB(p-NF-κB)in liver tissue.The serum metabolites of mice in each group were analyzed using ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry(UPLC-Q-TOF-MS).Results The content of gentiopicroside in Gentianae Radix et Rhizoma and Gentianae Radix et Rhizoma processed with wine was 3.84%and 3.12%,respectively,and the total polysaccharide content was 3.27%and 5.03%.Compared with model group,the liver injury of mice in gentiopicroside in Gentianae Radix et Rhizoma high-dose group and Gentianae Radix et Rhizoma processed with wine high-dose group was significantly improved,the a

关 键 词：龙胆 酒炙 急性肝损伤 代谢组学 龙胆苦苷 甘油磷脂代谢 

分 类 号：R285.5[医药卫生—中药学]