基于ERK/NF-κB/COX-2信号通路研究珠子参总皂苷改善对乙酰氨基酚致小鼠肝损伤的作用机制  

Mechanism of total saponins of Panax japonicus against liver injury induced by acetaminophen in mice based on ERK/NF-κB/COX-2 signaling pathway

作　　者：石孟琼[1] 张继红[2] 周刚[2] 郭巍 刘英[1] 包中文 陈茂华[2] 谈发明[2] 胡慧泉 汪鋆植[1] 任剑峡 唐海明 SHI Meng-qiong;ZHANG Ji-hong;ZHOU Gang;GUO Wei;LIU Ying;BAO Zhong-wen;CHEN Mao-hua;TAN Fa-ming;HU Hui-quan;WANG Jun-zhi;REN Jian-xia;TANG Hai-ming(Basic Medical College of China Three Gorges University&Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,Yichang 443002,China;Traditional Chinese Medicine College of China Three Gorges University&Hubei Clinical Research Center for Functional Digestive Diseases of Traditional Chinese Medicine,Yichang 443002,China;Pharmacy Department of Ezhou Second Hospital,Ezhou 436000,China;Yichang Second People′s Hospital,Yichang 443002,China;Hubei Heng′an Fulin Pharmaceutical Industry Group Co.,Ltd.,Yichang 443199,China)

机构地区：[1]三峡大学基础医学院&肿瘤微环境与免疫治疗湖北省重点实验室,湖北宜昌443002 [2]三峡大学中医医院&湖北省功能性消化系统疾病中医临床医学研究中心,湖北宜昌443002 [3]湖北鄂州二医院药剂科,湖北鄂州436000 [4]宜昌市第二人民医院,湖北宜昌443002 [5]湖北恒安芙林药业股份有限公司,湖北宜昌443199

出　　处：《中国中药杂志》2024年第10期2585-2596,共12页China Journal of Chinese Materia Medica

基　　金：湖北省科技厅重点研发大健康计划项目(2022BCE017);湖北省科技厅自然科学基金项目(2022CFB357,2022CFB427);湖北省卫生健康委员会中医药重点项目(ZY2023Z015);湖北卫生健康委员会卫生健康科研项目(WJ2023M153);湖北省老年胃肠癌精准防治临床医学研究中心开放基金项目(2022EGC-04);湖北省宜昌市科学技术局医疗卫生研究项目(A23-1-061)。

摘　　要：研究珠子参总皂苷(total saponins of Panax japonicus,TSPJ)对对乙酰氨基酚(acetaminophen,APAP)诱导小鼠肝损伤的影响及作用机制。将雄性昆明小鼠随机分为空白组、TSPJ组(200 mg·kg^(-1),ig)、模型组、APAP+TSPJ低剂量组(50 mg·kg^(-1),ig)、APAP+TSPJ中剂量组(100 mg·kg^(-1),ig)、APAP+TSPJ高剂量组(200 mg·kg^(-1),ig)和APAP+N-乙酰半胱氨酸组(200 mg·kg^(-1),ip)。给药组每天ig或ip相应的药物,每天1次,连续14 d。末次给药1 h后,除空白组和TSPJ组外,各组小鼠均灌胃给予500 mg·kg^(-1) APAP,24 h后收集小鼠血清与肝脏组织进行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)、活性氧(reactive oxygen species,ROS)、肿瘤坏死因子(tumor necrosis factor,TNF)-α、白细胞介素(interleukin,IL)-1β、环氧合酶(cyclooxygenase,COX)-2、IL-6、IL-4、IL-10及肝组织中乳酸脱氢酶(lactate dehydrogenase,LDH)、谷胱甘肽(glutathione,GSH)、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、总抗氧化能力(total antioxidant capacity,T-AOC)、丙二醛(malondialdehyde,MDA)、髓过氧化物酶(myeloperoxidase,MPO)的水平,苏木精-伊红染色观察肝组织形态学改变;实时定量PCR法测定肝组织中淋巴细胞抗原6G(lymphocyte antigen 6G,Ly6G)、半乳糖凝集素3(galectin 3,Mac-2)、TNF-α、IL-1β、COX-2、IL-6、IL-4、IL-10 mRNA表达,Western blot法测定肝组织中Ly6G、Mac-2、细胞外调节蛋白激酶(extracellular regulated protein kinase,ERK)、磷酸化细胞外信号调节激酶(phosphorylated extracellular regulated protein kinase,p-ERK)、COX-2、核因子κB抑制因子α(inhibitor of nuclear factorκB protein α,IκBα)、磷酸化核因子κB抑制因子α(phosphorylated inhibitor of nuclear factorκB protein α,p-IκBα)、胞浆和胞核中核因子κB亚基p65(nuclear factorκB subunit p65,NF-κB p65)蛋白表达。结果显示,TSPJ可显著降低APAP诱导小鼠肝�This study investigated the effects and mechanisms of total saponins of Panax japonicus(TSPJ) against liver injury induced by acetaminophen(APAP).Male Kunming mice were randomly divided into a blank control group,TSPJ group(200 mg·kg~(-1),ig),model group,APAP+ TSPJ low-dose group(50 mg·kg~(-1),ig),APAP+ TSPJ medium-dose group(100 mg·kg~(-1),ig),APAP+ TSPJ high-dose group(200 mg·kg~(-1),ig),and APAP+ N-acetyl-L-cysteine group(200 mg·kg~(-1),ip).The administration group received the corresponding medications via ig or ip once a day for 14 consecutive days.After the last administration for one hour,except for the blank control group and TSPJ group,all groups of mice were given 500 mg·kg~(-1) APAP by gavage.After 24 hours,mouse serum and liver tissue were collected for serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),reactive oxygen species(ROS),tumor necrosis factor alpha(TNF-α),interleukin-1 beta(IL-1β),cyclooxygenase-2(COX-2),IL-6,IL-4,IL-10,as well as lactate dehydrogenase(LDH),glutathione(GSH),superoxide dismutase(SOD),catalase(CAT),total antioxidant capacity(T-AOC),malondialdehyde(MDA),and myeloperoxidase(MPO) liver tissue.Hematoxylin-eosin staining was used to observe the morphological changes of liver tissue.The mRNA expression levels of lymphocyte antigen 6G(Ly6G),galectin 3(Mac-2),TNF-α,IL-1β,COX-2,IL-6,IL-4,and IL-10 in liver tissue were determined by quantitative real-time polymerase chain reaction(PCR).Western blot was utilized to detect the protein expression levels of Ly6G,Mac-2,extracellular regulated protein kinases(ERK),phosphorylated extracellular regulated protein kinases(p-ERK),COX-2,inhibitor of nuclear factor κB protein α(IκBα),phosphorylated inhibitor of nuclear factor κB protein α(p-IκBα),and nuclear factor-κB subunit p65(NF-κB p65) in cytosol and nucleus in liver tissue.The results manifested that TSPJ dramatically reduced liver coefficient,serum ALT,AST,ROS,TNF-α,IL-1β,IL-6,and COX-2 levels,LDH,MPO,and MDA contents in liver tissue,and mRNA expression

关 键 词：珠子参总皂苷 对乙酰氨基酚 氧化损伤 炎症反应 细胞外调节蛋白激酶/核因子κB/环氧合酶-2信号通路 

分 类 号：R285.5[医药卫生—中药学]