基于NF-κB/NLRP3通路和代谢组学的复方枳葛饮对慢性酒精性肝损伤的保护作用及机制研究  

Protective effect and mechanism of Fufang Zhige Yin on chronic alcoholic liver injury based on NF-κB/NLRP3 pathway and metabolomics

作　　者：王浩 史雯馨 邵仕娟 周锦航 陈运中 WANG Hao;SHI Wenxin;SHAO Shjuan;HOU Jinhang;CHEN Yunzhong(School of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Provincial Chinese Medicine Health Food Engineering Research Center,Wuhan 430065,China;Hubei Shizhen Laboratory,Wuhan 430065,China)

机构地区：[1]湖北中医药大学药学院,武汉430065 [2]湖北省中药保健食品工程技术研究中心,武汉430065 [3]湖北时珍实验室,武汉430065

出　　处：《中华中医药杂志》2024年第5期2297-2307,共11页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖北省中央引导地方科技发展专项(No.2019ZYYD069)。

摘　　要：目的:探讨复方枳葛饮对慢性酒精性肝损伤的治疗作用,并通过核因子κB(NF-κB)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)通路和代谢组学的方法探究其发挥作用的机制。方法:将小鼠分为空白组,模型组,联苯双酯组,复方枳葛饮水提物高、低剂量组,醇提物高、低剂量组。除空白组外,其余各组予以酒精灌胃制备慢性酒精性肝损伤模型,并同时给予相应的药物治疗,8周后,检测小鼠肝脏谷草转氨酶(AST)、谷丙转氨酶(ALT)、超氧化物歧化酶(SOD)、丙二醛(MDA)、甘油三酯(TG)、总胆固醇(TC)指标,HE染色来观察小鼠肝脏病理改变,ELISA法检测小鼠血清炎症因子白介素-18(IL-18)、肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)水平,qRT-PCR检测小鼠肝脏TNF-α、IL-6、IL-1β mRNA表达,Western Blot检测肝脏NF-κB和NLRP3蛋白表达,采用液相色谱-质谱联用仪(LC-MS)非靶向代谢组学技术分析血清及肝脏内源性代谢产物变化及复方枳葛饮的作用途径。结果:与模型组比较,复方枳葛饮水提物和醇提物均能显著降低小鼠肝脏ALT、AST、TC、TG,血清TNF-α水平、肝脏炎症基因IL-1β mRNA水平表达、肝脏NF-κB蛋白及NLRP3蛋白的表达(P<0.05,P<0.01);水提物还能显著升高小鼠SOD活力,显著降低小鼠肝脏MDA、TNF-α mRNA表达(P<0.05,P<0.01);醇提物还能显著降低血清IL-18水平(P<0.05);血清代谢组学筛选出10种与醇提物治疗酒精性肝损伤相关的血清代谢物,其中可上调6个、下调4个,12种与水提物治疗酒精性肝损伤相关的血清代谢物,其中可上调6个、下调6个;肝脏代谢组学筛选出21种与水提物治疗酒精性肝损伤相关的肝脏代谢物,其中可上调13个、下调8个,涉及的代谢途径主要为抗坏血酸和醛酸代谢、嘌呤代谢、精氨酸生物合成等途径;筛选出21种与醇提物治疗酒精性肝损伤相关的肝脏代谢物,其中可上调2个、下调19个,涉及的代谢通路主要为淀粉和�Objective: To explore the therapeutic effect of Fufang Zhige Yin on alcoholic liver injury and its mechanism based on NF-κB/NLRP3 pathway and metabolomics. Methods: The mice were divided into blank group, model group, group, high and low dose of Fufang Zhige Yin water extract, and high and low dose of Fufang Zhige Yin alcohol extract. In addition to the blank group, the rest of the groups were given alcohol gavage to prepare the alcoholic liver injury model, and at the same time,the corresponding drug treatment was given. After 8 weeks, the liver ALT, AST, TC, TG, SOD, MDA indexes were detected in mice, HE staining was used to observe the pathological changes in the liver of the mice, and the serum levels of inflammatory factors TNF-α, IL-1β, and IL-18 were detected in the mouse serum by ELISA method, and the levels of the inflammatory factors TNF-α, IL-1β, and IL-18 were detected in the liver of the mice by qRT-PCR to detect mRNA expression of TNF-α,IL-6, IL-1β genes in mouse liver, Western Blot was used to detect NF-κB and NLRP3 protein expression in liver, and LC-MS non-targeted metabolomics to analyze the changes of endogenous metabolites and pathways in serum and liver. Results: Compared with the model group, both the extracts of the Fufang Zhige Yin water and the alcoholic extracts significantly reduced the hepatic ALT, AST, TC, TG, serum TNF-α levels, hepatic expression of inflammatory gene IL-1β mRNA levels, hepatic expression of NF-κB protein and NLRP3 protein in mice(P<0.05, P<0.01);the aqueous extracts also significantly elevated the SOD activity of mice and significantly reduced the mouse liver MDA and TNF-α mRNA expression(P<0.05, P<0.01);the alcoholic extract also significantly reduced serum IL-18 levels(P<0.05);serum metabolomics screened for 10 metabolites associated with alcoholic extract for the treatment of alcoholic liver injury, of which 6 could be up-regulated and 4 could be down-regulated, and 12aqueous extracts for the treatment of alcoholic liver injury metabolites, of which 6

关 键 词：复方枳葛饮 酒精性肝损伤 NF-κB/NLRP3 血清代谢组学 肝脏代谢组学 机制 

分 类 号：R285.5[医药卫生—中药学]