短穗兔耳草中3个化合物对大鼠慢性酒精性肝损伤合并痛风性关节炎的作用及其机制研究  

The Effect and Mechanism of Three Compounds in Lagotis brachystachya Maxim on Chronic Alcoholic Liver Injury Combined with Gouty Arthritis in Rats

作　　者：朱继孝 周琳 张初玲 熊雯雯[2] 程虹毓 ZHU Jixiao;ZHOU Lin;ZHANG Chuling;XIONG Wenwen;CHENG Hongyu(Research Center for Traditional Chinese Medicine Resources and Ethnic Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004 Jiangxi,China;Center for Public Health Education and Health Services,Jiangxi University of Chinese Medicine,Nanchang 330004 Jiangxi,China)

机构地区：[1]江西中医药大学中药资源与民族药研究中心,江西南昌330004 [2]江西中医药大学公共卫生教育与健康服务中心,江西南昌330004

出　　处：《中药新药与临床药理》2024年第6期822-831,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82060757);国家重点研发计划项目(2019YFC1712302);江西省卫生计生委中医药科研项目(2018A390,2019A258)。

摘　　要：目的通过建立大鼠慢性酒精性肝损伤合并痛风性关节炎模型,研究槲皮素、松果菊苷和芹菜素对p38MAPK/JNK、TLR4/MyD88/NF-κB和NLRP3信号通路的影响,探讨短穗兔耳草中单体化合物抗慢性酒精性肝损伤合并痛风性关节炎的作用机制。方法将80只大鼠随机分为正常组,模型组,联苯双酯组(100 mg·kg^(-1)),秋水仙碱组(0.3 mg·kg^(-1))及槲皮素(50、25 mg·kg^(-1))、松果菊苷(50、25 mg·kg^(-1))、芹菜素(50、25 mg·kg^(-1))各剂量组。每日上午按10 mL·kg^(-1)给药,下午以4 mL·kg^(-1)开始梯度给56度红星二锅头,每周增加2 mL·kg^(-1),直至10 mL·kg^(-1)保持,连续灌胃8周。于灌胃第53天复制痛风性关节炎模型,检测各时间段足趾容积。第8周末次给药后取血,检测血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)水平、总胆固醇(TC)、甘油三酯(TG);酶联免疫吸附测定(ELISA)检测大鼠血清中白细胞介素1β(IL-1β);蛋白免疫印迹法(Western Blot)检测肝脏和滑膜中p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)、c-Jun氨基末端激酶(JNK)和磷酸化c-Jun氨基末端激酶(p-JNK)、Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核因子κB(NF-κB)、磷酸化核因子κB(p-NF-κB)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)蛋白的表达。结果与正常组比较,模型组各时间段大鼠足肿胀度均明显升高(P<0.05,P<0.01);血清中ALT、AST、ALP、TC、TG、IL-1β水平均明显升高(P<0.01);肝组织和滑膜组织中p-p38MAPK、p-JNK、MyD88、p-NF-κB、TLR4、NLRP3蛋白表达水平均有不同程度升高(P<0.01)。与模型组比较,各给药组在24 h足肿胀度呈现明显下降趋势(P<0.01);各给药组ALT、TC水平有不同程度下降(P<0.05,P<0.01);除松果菊苷低剂量组外,各给药组AST、ALP水平有不同程度下降(P<0.01);除槲皮素高剂量组外,各给药组TG水平有不同程度下降(P<0.01);各组大鼠肝脏组织Objective To study the effects of monomers in Lagotis brachystachya Maxim including quercetin,echinacoside,and apigenin on the p38MAPK/JNK,TLR4/MyD88/NF-κB,and NLRP3 signaling through establishing a rat model of chronic alcoholic liver injury combined with gouty arthritis,and to explore the mechanism of the above-mentioned monomers in the treatment of chronic alcoholic liver injury combined with gouty arthritis.Methods Eighty rats were randomly divided into normal group,model group,bifendate group(100 mg·kg^(-1)),colchicine group(0.3 mg·kg^(-1)),and different dose groups of quercetin(50,25 mg·kg^(-1)),echinacoside(50,25 mg·kg^(-1)),and apigenin(50,25 mg·kg^(-1)).The rats were administered at a dose of 10 mL·kg^(-1)per day in the morning and administered with 56-degree Red Star Erguotou at a dose of 4 mL·kg^(-1)in the afternoon,with gradual increasement of 2 mL·kg^(-1)per week until a final dose of 10 mL·kg^(-1).The rats were continuously fed by gavage for 8 weeks.Gouty arthritis model was induced at the 53th day of alcohol-based feeding.Toe volume was measured at difference time.At the end of the 8th week,blood was collected after the last administration.Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP)levels,total cholesterol(TC)and triglycerides(TG)were measured.Enzyme-Linked Immunosorbent Assay(ELISA)was used to detect interleukin-1β(IL-1β)in rat serum.Western Blot was used to detect the expression of p38 mitogen-activated protein kinase(p38MAPK),phosphorylated p38MAPK(p-p38MAPK),c-Jun N-terminal kinase(JNK),phosphorylated JNK(p-JNK),Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),nuclear factor-κB(NF-κB),phosphorylated NF-κB(p-NF-κB),and NOD-like receptor protein domain containing 3(NLRP3)in liver and synovial tissues.Results Compared with the normal group,the toe swelling degree in the model group was significantly increased at difference time(P<0.05,P<0.01).The levels of ALT,AST,ALP,TC,TG,and IL-1βin serum were significantly in

关 键 词：短穗兔耳草 槲皮素 松果菊苷 芹菜素 慢性酒精性肝损伤 痛风性关节炎 p38MAPK/JNK TLR4/MyD88/NF-κB NLRP3 大鼠 

分 类 号：R285.5[医药卫生—中药学]