槐果碱改善对乙酰氨基酚诱导的小鼠急性肝损伤机制研究

Mechanism of Sophocarpine in Improving Acetaminophen-Induced Acute Liver Injury in Mice

目的:研究槐果碱在对乙酰氨基酚(APAP)诱导的小鼠急性肝损伤模型上的影响和作用。方法:通过网络药理学在PubChem数据库获取槐果碱化合物的结构,采用PharmMapper和Swiss Target Prediction数据库筛选药物靶点,利用GeneCards、OMIM数据库进行疾病靶点预测,将药物和疾病的交集导入STRING数据库构建PPI网络,预测槐果碱治疗APAP诱导的小鼠急性肝损伤作用靶点及通路。采用C57BL/6小鼠建立APAP诱导的急性肝损伤模型,同时用槐果碱干预小鼠,通过观察肝脏表观、血清生化检测、肝脏HE染色、肝脏F4/80免疫组化染色和Tunel凋亡染色评价小鼠肝损伤情况。通过Western blot验证槐果碱治疗APAP诱导的小鼠急性肝损伤的作用机制。结果:网络药理学获得槐果碱治疗急性肝损伤91个交集靶点,涉及磷脂酰肌醇3-激酶/蛋白激酶B (PI3K-AKT)等信号通路。与模型对照组相比,槐果碱45、60 mg/kg组可明显降低APAP诱导的急性肝损伤小鼠ALT、AST活力(P<0.01),显著改善肝组织病理损伤程度、减少巨噬细胞浸润,并且明显减少肝细胞凋亡(P<0.01),Western blot结果显示槐果碱60 mg/kg可明显上调p-PI3K、p-AKT蛋白表达(P<0.05或P<0.01)。结论:槐果碱通过抗炎、抗凋亡作用来减轻APAP诱导的肝损伤,作用机制与激活PI3K/AKT通路相关。

Objective:To study the effects of sophocarpine on acute liver injury induced by acetaminophen(APAP)in mice.Methods:Network pharmacology was used to obtain the structure of sophocarpine through the PubChem database,and drug targets were screened in PharmMapper and Swiss Target Prediction databases.GeneCards and OMIM databases were used for the prediction of disease targets.The intersection of the drug and disease was imported into the STRING database and protein-protein interaction(PPI)network was constructed to predict the possible targets and related pathways of sophocarpine in the treatment of APAP-induced acute liver injury.Then C57BL/6 mice were used to establish the model of APAP-induced acute liver injury.Meanwhile,mice were treated with sophocarpine.The degree of liver injury of mice was evaluated via liver appearance observation,serum biochemical detection,HE staining,F4/80 immunohistochemical staining,and Tunel apoptosis staining of liver tssues.Western blot was used to verify the underlying mechanism of sophocarpine in the treatment of APAP-induced acute liver injury in mice.Results:Ninety-one intersection targets of sophocarpine in the treatment of acute liver injury were obtained by network pharmacology,involving phosphatidylinositol 3-kinase-protein kinase B(PI3K-AKT)and other signaling pathways.Compared with the model control group,the sophocarpine groups(45 and 60 mg/kg)markedly blunted the activities of ALT and AST in APAP-induced acute liver injury mice(P<0.01),improved the degree of pathological injury of liver tissues,reduced the infiltration of macrophages,and significantly decreased the apoptosis of liver tissues(P<0.01).The results of Western blot showed that 60 mg/kg sophocarpine markedly up-regulated the protein expression of p-PI3K and p-AKT(P<0.05 or P<0.01).Conclusion:Sophocarpine can alleviate APAP-induced liver injury through anti-inflammatory and anti-apoptotic effects,and its mechanism of action may be related to the activation of the PI3K/AKT pathway.