本文研究诸葛菜种子水提物对对乙酰氨基酚（APAP）诱导小鼠急性肝损伤的保护作用。健康雄性C57 BL/6J小鼠被随机分为7组：正常组，模型组，双环醇阳性对照组（200 mg/kg），葵花护肝片阳性对照组（350 mg/kg），诸葛菜种子水提物低（125 mg/kg）、中（250 mg/kg）、高（500 mg/kg）剂量组。各组均灌胃给药，1次/d，连续灌胃给药4 d，除正常组外其余各组均腹腔注射APAP溶液（450 mg/kg）建立急性肝损伤模型，处死小鼠。检测血清中丙氨酸氨基转移酶（ALT），谷草转氨酶（AST）活性与胆红素（TBIL）水平；肝组织中超氧化物歧化酶（SOD）、丙二醛（MDA）、还原型谷胱甘肽（GSH）；HE染色观察小鼠肝组织病理变化；蛋白印迹法检测磷酸化腺苷酸活化蛋白激酶（p-AMPK）、磷酸化AKT蛋白（p-AKT）、磷酸化葡萄糖合成激酶3β（p-GSK-3β）、磷酸化c-Jun氨基末端激酶( p-JNK)、B淋巴细胞瘤-2基因（Bcl-2）、Bcl-2相关X蛋白（Bax）、活化半胱胺酸蛋白酶蛋白-9抗体（Caspase-9）、活化半胱胺酸蛋白酶蛋白-3抗体（Caspase-3）蛋白的表达。与模型组相比，诸葛菜各剂量组均能明显改善肝脏病理变化，中剂量组ALT，AST活性分别降低了48.73%、59.11%，高剂量组分别降低了71.38%、59.33%，中剂量组SOD和GSH分别上升了45.05%、219.45%，高剂量组SOD和GSH分别上升了48.68%和232.80%，同时可致肝组织中p-JNK、Bax、Caspase-3、Caspase-9蛋白表达水平显著减少，Bcl-2、p-AMPK、p-AKT、p-GSK-3β蛋白表达水平显著增加。其机制可能与AMPK/AKT/GSK-3β信号通路被激活有关。

The antioxidant capacity of water extracts of Orychophragmus violaceus Seeds and the protective effects on liver injury induced by acetaminophen in mice were investigated. C57BL/6J male mice were randomly divided into seven groups: normal group, model group, bicycle control group (200 mg/kg), Kuihua Hugan tablets control group (350 mg/kg), water extracts of Orychophragmus violaceus seeds low-dose group (125 mg/kg), water extracts of Orychophragmus violaceus seeds middle-dose group (250 mg/kg), and water extracts of Orychophragmus violaceus seeds high-dose group (500 mg/kg). The mice were intragastrically administered with drugs or distilled water according to the body weight once a day. Four days later, all of the groups except for the normal group were treated with APAP (450 mg/kg) through intraperitioneal injection to establish the acute liver injury model. The mice were killed. The levels of glutamic-pyruvic transaminase, glutamic oxalacetic transaminase and total bilirubin in serum were detected. What’s more, the activities of superoxide dismutase and the level of glutathione and malondiadehyde in liver tissue were detected. HE staining was used to observe the pathological changes of liver tissue in mice. Western blot was used to detect the expressions of p-AMPK, p-AKT, p-GSK-3β, p-JNK, Bcl-2, Bax, Caspase-9 and Caspase-3. Compared with the model group, water extracts of Orychophragmus violaceus seeds could significantly improve the pathological conditions of liver, and the ALT and AST activities in the middle-dose group were reduced by 48.73% and 59.11%, respectively; and the high-dose group were reduced by 71.38% and 59.33%, respectively, in serum of APAP induced liver injury mice. The activities of SOD and GSH were increased by 45.05% and 219.45% respectively in the middle dose group and 48.68% and 232.80% respectively in the high dose group. Meanwhile, water extracts of Orychophragmus violaceus seeds down-regulated the expressions of p-JNK, Bax, Caspase-3, and Caspase-9 (p<0.05) and up-regulated the expressions of Bcl-2, p-AMPK, p-AKT and p-GSK-3β (p<0.05). It suggested that water extracts of Orychophragmus violaceus seeds has protective effect against APAP induced hepatocyte apoptosis by up-regulating the AMPK/AKT/GSK-3β signaling pathway.

北京市自然科学基金青年科学基金项目（7164281）