基于MAPK信号转导通路探究异甘草酸镁对刀豆蛋白A诱导小鼠急性肝损伤的保护机制  

Magnesium isoglycyrrhizinate against concanavalin A-induced acute liver injury in mice based on MAPK signaling pathway

作　　者：王俊文[1] 范子豪 田原 徐玲 高耀 曹亚玲 潘桢桢 张向颖[2] 宋岩 任锋[2] Wang Junwen;Fan Zihao;Tian Yuan;Xu Ling;Gao Yao;Cao Yaling;Pan Zhenzhen;Zhang Xiangying;Song Yan;Ren Feng(Department of Clinical Laboratory,Beijing Chuiyangliu Hospital,Beijing 100022,China;Beijing Institute of Hepatology,Beijing Youan Hospital,Capital Medical University,Beijing 100069,China)

机构地区：[1]北京市垂杨柳医院检验科,北京100022 [2]首都医科大学附属北京佑安医院北京肝病研究所,北京100069

出　　处：《中国肝脏病杂志（电子版）》2023年第4期47-53,共7页Chinese Journal of Liver Diseases：Electronic Version

基　　金：国家自然科学基金项目(81770611、82002243);北京自然科学基金和北京市教委联合资助重点项目(KZ202010025035);首都卫生发展科研专项重点攻关项目(首发2020-1-1151,首发2021-1G-2181);北京市科技计划“首都临床诊疗技术研究及示范应用”专项课题(Z191100006619096、Z191100006619097);北京市优秀人才培养项目(2018000021469G289);北京市医院管理中心“青苗”计划专项经费资助(QML20201702)。

摘　　要：目的 探讨异甘草酸镁(magnesium isoglycyrrhizinate,MgIG)在刀豆蛋白A(concanavalin A,Con A)诱导的小鼠急性肝损伤中的作用及机制。方法 按照简单随机分组法将20只无特定病原体(specific pathogen free,SPF)级Balb/c小鼠分为正常对照组(4只)、MgIG组(4只)、Con A组(6只)、Con A+MgIG干预组(6只)。小鼠Con A(25 mg/kg)尾静脉注射12 h,构建急性肝损伤模型,干预组提前1 h给予MgIG(30 mg/kg)腹腔注射。检测丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天门冬氨酸转氨酶(aspartate aminotransferase,AST)水平、白细胞介素(interleukin,IL)-1β、IL-6、肿瘤坏死因子(tumor necrosis factor α,TNF-α)、干扰素诱生蛋白10(interferon-inducibleprotein-10,IP-10)水平,检测IL-6、IL-1β、TNF-α、IP-10的mRNA相对表达量。体外实验中小鼠腹腔单核巨噬细胞用脂多糖(lipopolysaccharide,LPS,200 ng/ml)分别处理30 min、1 h、2 h和4 h,干预组小鼠腹腔单核巨噬细胞用MgIG(25μg/ml)预处理1h,检测IL-6、IL-β、TNF-α、IP-10炎症因子及磷酸化-p38丝裂原活化蛋白激酶(phospho-p38 mitogen activited protein kinase, p-p38)、磷酸化-cJun氨基末端激酶(phospho-c-JunN-terminalkinases,p-JNK)、磷酸化-细胞外调节蛋白激酶(phospho-extracellular regulated protein kinases,p-Erk)蛋白表达。结果 与Con A组相比,Con A+MgIG干预病理切片炎性细胞浸润明显减少,血清炎症因子[IL-6:(10695.71±4861.94)pg/ml vs (27650.88±5701.79)pg/ml;IL-1β:(13.37±8.18)pg/ml vs (56.55±9.29)pg/ml;IP-10:(3298.43±534.95)pg/ml vs (7413.38±1497.78)pg/ml;TNF-α(63.27±13.97)pg/ml vs (97.06±21.26)pg/ml]及mRNA相对表达量(IL-6:5.23±1.63 vs 16.06±4.55;IL-1β:0.88±0.45 vs 5.44±0.94;IP-10:126.24±29.54vs 454.40±114.81;TNF-α:9.55±2.75 vs 16.46±3.98)均显著降低(P均<0.05)。体外实验表明,与LPS诱导的模型组相比,MgIG干预组p-p38、p-Jnk、p-Erk蛋白水平明显降低,同时炎症因子mRNA相对表达量(IL-6:3627.91±1491.16 vs 6630.40±1149.59;IL-1β:259.Objective To investigate the role and mechanism of magnesium isoglycyrrhizinate(MgIG)in concanavalin A(Con A)-induced acute liver injury.Methods In vivo experiments,20 specific pathogen free(SPF)Balb/c mice were randomly divided into normal control group(4 cases),MgIG control group(4 cases),Con A model group(6 cases)and Con A+MgIG intervention group(6 cases)by simple randomization grouping method.The mice were injected with Con A(25 mg/kg)intravenously into the tail vein for 12 h to establish an acute liver injury model,and the intervention mice were injected with MgIG(30 mg/kg)intraperitoneally 1 h earlier.Levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),interleukin(IL)-1β,IL-6,tumor necrosis factorα(TNF-α)and interferon-inducible protein-10(IP-10)were detected.The relative mRNA expression levels of IL-6,IL-1β,TNF-αand IP-10 were also detected.For in vitro experiments,the mice peritoneal mononuclear macrophages were treated with lipopolysaccharide(LPS,200 ng/ml)for 30 min,1 h,2 h and 4 h,and the mice peritoneal mononuclear macrophages in the intervention group were pretreated with MgIG(25μg/ml)for 1 h,and the expression of inflammatory factors(IL-6,IL-β,TNF-α,IP-10)and related proteins[phospho-p38 mitogen activited protein kinase(p-p38),phosphoc-JunN-terminal kinases(p-JNK),phospho-extracellular regulated protein kinases(p-Erk)]were detected.Results Compared with those in Con A model group,the inflammatory cell infiltration of mice in Con A+MgIG intervention group reduced significantly,serum inflammatory factors[IL-6:(10695.71±4861.94)pg/ml vs(27650.88±5701.79)pg/ml;IL-1β:(13.37±8.18)pg/ml vs(56.55±9.29)pg/ml;IP-10:(3298.43±534.95)pg/ml vs(7413.38±1497.78)pg/ml;TNF-α(63.27±13.97)pg/ml vs(97.06±21.26)pg/ml]and relative mRNA expression levels(IL-6:5.23±1.63 vs 16.06±4.55;IL-1β:0.88±0.45 vs 5.44±0.94;IP-10:126.24±29.54 vs 454.40±114.81;TNF-α:9.55±2.75 vs 16.46±3.98)also reduced significantly(all P<0.05).Results of in vitro experiments showed that compared with thos

关 键 词：急性肝损伤 异甘草酸镁 刀豆蛋白A 炎症反应 丝裂原活化蛋白激酶信号转导通路 

分 类 号：R575[医药卫生—消化系统]