楮实子通过抑制内质网应激途径保护APAP 诱导的小鼠肝损伤  

Broussonetiae Fructus Protects Against APAP-induced Liver Injury in Mice by Inhibiting Endoplasmic Reticulum Stress Pathway

作　　者：高晶淼 王婷婷 彪雅宁 顾亚茹 张睦清[3] 王茜 张一昕 GAO Jingmiao;WANG Tingting;BIAO Yaning;GU Yaru;ZHANG Muqing;WANG Xi;ZHANG Yixin(School of Pharmacy,International Joint Research Center on Resource Utilization and Quality Evaluation of Traditional Chinese Medicine(TCM)of Hebei Province,Application Technology Research and Development Center of TCM in Hebei Universities,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Cangzhou People's Hospital,Cangzhou 061000,China;Hebei Provincial Hospital of TCM,Shijiazhuang 050011,China)

机构地区：[1]河北中医学院药学院,河北省中药资源利用与质量评价国际联合研究中心,河北省高校中药组方制剂应用技术研发中心,石家庄050200 [2]沧州市人民医院,沧州061000 [3]河北省中医院,石家庄050011

出　　处：《中国实验方剂学杂志》2022年第16期66-73,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(N081503268);河北省高等学校科学技术研究青年拔尖计划项目(BJ2016038);河北省中医药管理局科研计划项目(2020308,2022362)。

摘　　要：目的:探讨楮实子基于内质网应激途径防治对乙酰氨基酚(APAP)致药物性肝损伤(DILI)的作用机制。方法:将60只C57BL/6N小鼠随机分为6组:正常组、模型组、水飞蓟宾组(3.4 g·kg^(-1))和楮实子高、中、低剂量组(3.0、1.5、0.75 g·kg^(-1)),每组10只。采用APAP(800 mg·kg^(-1))灌胃的方法造模,同时给药治疗,连续10 d。检测血清中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、直接胆红素(DBIL)的含量或活性;苏木素-伊红(HE)染色观察肝组织病理形态变化;透射电镜观察肝细胞线粒体形态变化;比色法检测血清和肝组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化能力(T-AOC)、谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、谷胱甘肽过氧化物酶(GSH-Px)、三磷酸腺苷(ATP)的活性或含量;免疫荧光法检测肝组织中活性氧(ROS)的表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织葡萄糖调节蛋白78(GRP78)、CCAAT/增强子结合蛋白同源蛋白(CHOP)、c-Jun氨基末端激酶(JNK)的mRNA表达水平。结果:与正常组比较,模型组小鼠血清中ALT、AST、TBIL、DBIL活性或含量显著升高(P<0.01);MDA和GSSG含量显著升高(P<0.01),SOD、T-AOC、GSH、GSH-Px、ATP含量或活性显著降低(P<0.01);肝细胞出现肿胀、炎性浸润及片状坏死;肝细胞线粒体肿胀破裂;GRP78、CHOP和JNK mRNA表达均显著增强(P<0.01)。与模型组比较,各给药组小鼠血清中ALT、AST、TBIL、DBIL含量或活性明显降低(P<0.05,P<0.01);MDA和GSSG含量明显降低(P<0.05,P<0.01),SOD、T-AOC、GSH、GSH-Px、ATP含量或活性明显升高(P<0.05,P<0.01);肝细胞肿胀、炎性浸润及片状坏死减少;肝细胞线粒体恢复双层膜结构;GRP78、CHOP和JNK mRNA表达均明显减弱(P<0.05,P<0.01)。结论:楮实子对APAP肝损伤小鼠具有防治作用,可能与减轻内质网应激、降低体内氧化应激水平有关。Objective:To explore the mechanism of Broussonetiae Fructus(BF)in preventing and treating drug-induced liver injury(DILI)induced by acetaminophen(APAP)through the endoplasmic reticulum stress pathway.Method:Sixty C57BL/6N mice were randomly divided into normal group,model group,silybin group(3.4 g·kg^(-1)),and high-,medium-and low-dose BF groups(3.0,1.5,0.75 g·kg^(-1)),with 10 mice in each group.The DILI model was induced by intragastric administration of APAP at 800 mg·kg^(-1),and drugs were administered simultaneously for 10 consecutive days.The serum contents or activities of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),and direct bilirubin(DBIL)were measured.Hematoxylin-eosin(HE)staining was performed to observe the pathological changes in liver tissues.The morphological changes in liver mitochondria were observed by transmission electron microscopy.The activities or content of superoxide dismutase(SOD),malondialdehyde(MDA),total antioxidant capacity(T-AOC),glutathione(GSH),glutathione disulfide(GSSG),glutathione peroxidase(GSH-Px),and adenosine triphosphate(ATP)in the serum and liver tissues were detected by the colorimetric method.The expression of reactive oxygen species(ROS)in liver tissues was detected by immunofluorescence.The gene expression of glucose-regulated protein 78(GRP78),CCAAT/enhancer-binding protein homologous protein(CHOP),and c-Jun N-terminal kinase(JNK)in liver tissues was detected by Real-time quantitative polymerase chain reaction(PCR).Result:Compared with the normal group,the model group showed increased serum activities or content of ALT,AST,TBIL,and DBIL(P<0.01),increased MDA and GSSG contents(P<0.01),decreased contents or activities of SOD,T-AOC,GSH,GSH-Px,and ATP(P<0.01),swollen hepatocytes with inflammatory infiltration and lamellar necrosis,swollen and broken mitochondria of hepatocytes,and increased mRNA expression of GRP78,CHOP,and JNK(P<0.01).Compared with the model group,the groups with drug intervention showed decreased serum content

关 键 词：楮实子 对乙酰氨基酚 药物性肝损伤 内质网应激 

分 类 号：R2-0[医药卫生—中医学] R33R256.4