目的 分析龙胆苦苷对高脂高糖饮食构建的非酒精性脂肪肝病（NAFLD）大鼠的干预作用及其机制。方法 60只SD大鼠按照随机数字法分为对照组、模型组、阳性药物多烯磷脂酰胆碱组及龙胆苦苷低、中、高剂量组，高脂高糖饮食12周建立NAFLD大鼠模型，龙胆苦苷低、中、高剂量组分别ig给予龙胆苦苷50、100、200 mg/kg，多烯磷脂酰胆碱组ig给予多烯磷脂酰胆碱23 mg/kg，对照组和模型组给予500 μL/kg生理盐水, 治疗8周后，收集各组大鼠血清，检测肝功能、血脂、血清氧化和抗氧化能力以及炎性因子相关指标，进行肝脏HE染色观察病理学改变，Western blotting和qRT-PCR检测肝脏组织中腺苷酸激活蛋白激酶α（AMPKα）和磷酸化的腺苷酸激活蛋白激酶α（p-AMPKα）的表达特点。结果 HE染色显示对照组大鼠肝脏细胞大小均一，细胞核呈均匀分布，模型组有明显脂肪空泡，且有一定的炎症反应浸润，与模型组相比，多烯磷脂酰胆碱和龙胆苦苷均有明显的改善作用，特别是龙胆苦苷中剂量和高剂量组已明显好转，但与对照组仍有一定差异；与对照组相比，模型组大鼠肝功能指标天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）、血脂高密度脂蛋白胆固醇（HDL-C）和低密度脂蛋白胆固醇（LDL-C）、氧化指标丙二醛（MDA）以及炎性因子白细胞介素-1（IL-1）和IL-6水平明显升高（P<0.05），抗氧化指标过氧化物歧化酶（SOD）活性明显下降（P<0.05），龙胆苦苷治疗后，AST、ALT、MDA、IL-1和IL-6明显降低（P<0.05），SOD明显升高（P<0.05），HDL-C和LDL-C也有一定的降低，但没有统计学意义，多烯磷脂酰胆碱则对上述指标均有明显的改善作用；与对照组相比，模型组p-AMPKα蛋白和AMPK mRNA明显降低（P<0.05）；使用多烯磷脂酰胆碱和龙胆苦苷治疗后，均明显升高（P<0.05），且龙胆苦苷3个剂量组有明显的剂量依赖性，同时中剂量和高剂量组要好于多烯磷脂酰胆碱组（P<0.05），而AMPKα蛋白表达在整个治疗过程中没有明显差异。结论 高脂高糖饮食12周构建的NAFLD大鼠有明显肝脏脂肪浸润，同时肝功能指标升高、血脂异常、炎性因子升高和抗氧化能力降低，龙胆苦苷则可以明显改善上述症状，这可能是龙胆苦苷增加大鼠肝脏组织中p-AMPKα蛋白表达实现的。

Objective To investigate the therapeutic effect of gentiopicroside has a on rats with non-alcoholic fatty liver disease (NAFLD) induced by high-fat and high-sucrose diet, and explore its mechanism. Methods After 10 d adaptive feeding, 60 male SD rats were randomly divided into the normal group, model group, gentiopicroside low, medium, high dose treatment groups, and positive drug polyeno phosphoryl choline (PPC) intervention group. Except for the normal group, the rats in other groups were received a high fat and glucose diet for 12 weeks to establish NAFLD model; After model successfully established, gentiopicroside low, medium, and high dosetreatment groups were given 50, 100, and 200 mg/(kg·d) gentiopicroside, PPC group was ig given 23 mg/(kg·d) PPC, and 500 μL/(kg·d) saline was given to the normal and model groups. After treated for eight weeks, the rats were sacrificed, and the serum was collected from rats to detect the liver function, blood lipid, serum oxidation, antioxidant capacity, and inflammatory factors. HE staining was used to observe pathological changes of liver. In addition, western blotting and qRT-PCR were used to detect the expression of AMPKα and p-AMPKα. Results HE staining showed that the size of liver cells in the normal group was uniform and the nuclei were evenly distributed, there were obvious vacuoles and a certain inflammatory reaction in the model group. Compared with the model group, gentiopicroside treatment group and PPC group (especially the gentiopicroside middle and high dose group) had a significant improvement, but there were still some differences compared with the normal group; Compared with the normal group, the AST, ALT, HDL-C, LDL-C, MDA, IL-1, and IL-6 in the model group were significantly increased (P < 0.05), the levels of AST, ALT, MDA, IL-1, and IL-6 were significantly decreased (P < 0.05), SOD was significantly increased (P < 0.05), and HDL-C and LDL-C were decreased in the model group (P > 0.05); The results of Western blotting and qRT-PCR showed that compared with the normal group, the expression of p-AMPKα protein and AMPKα mRNA in the model group was significantly decreased (P < 0.05). After gentiopicroside and PPC administration, they were significantly increased (P < 0.05), and gentiopicroside groups showed a significant dose-dependent manner, and the middle dose and high dose of gentiopicroside groups were better than the PPC group (P < 0.05), while the expression of AMPKα protein has no significant difference in each group (P < 0.05). Conclusion The NAFLD rats showed a obvious hepatic fat infiltration and dyslipidemia, the liver function index and inflammatory factors levels were elevated, and the anti-oxidant capacity was decreased. Gentiopicroside significantly improved above symptoms, which may be associated with the increased expression of p-AMPKα in liver tissue of NAFLD rats by gentiopicroside.

国家自然科学基金面上项目资助（81160492）