摘要
目的:探讨肝素参与野生型成纤维细胞生长因子1(FGF1WT)缓解对乙酰氨基酚(APAP)所致小鼠急性肝损伤(ALI)的作用机制。方法:实验分为两个批次,第一批C57BL/6J小鼠随机分为Control组、APAP组、APAP+FGF1WT组、APAP+FGF1^(WT)+Heparin组、APAP+FGF1改构体(FGF1^(ΔHBS))组以及APAP+FGF1^(ΔHBS)+Heparin组。第二批C57BL/6J小鼠随机分为Control组、Heparin组、APAP组、APAP+Heparin组。500 mg/kg剂量APAP造模1 h后,腹腔注射对应剂量FGF1WT(2 mg/kg)、FGF1^(ΔHBS)(2 mg/kg)和(或)肝素(1.5 mg/kg),造模6 h后处死小鼠,通过HE染色观察肝损伤,DHE荧光染色观察肝脏氧化应激,酶标仪法检测血清ALT和AST含量以及肝组织中超氧化物歧化酶和丙二醛含量,F4/80荧光染色观察肝脏炎症,qPCR检测过氧化氢酶(CAT)、核因子E2相关因子2(NRF2)、趋化因子配体5(CCL5)、TNFα和IL-6 mRNA水平,确定在肝素存在或不存在情况下,FGF1^(WT)和FGF1^(ΔHBS)对APAP小鼠ALI的缓解作用。结果:FGF1^(WT)对APAP所致小鼠ALI的治疗作用明显优FGF1^(△HBS),且肝素能明显增强FGF1^(△HBS)的作用,使其达到与FGF1^(WT)相类似效果(均P<0.05)。FGF1^(WT)明显抑制APAP所致小鼠肝组织氧化应激损伤和炎症,且FGF1^(WT)的抑制作用明显优FGF1^(△HBS)(均P<0.05)。肝素能明显增强FGF1^(△HBS)的抑制作用,使其达到与FGF1^(WT)相类似效果(均P<0.05)。FGF1^(WT)明显上调APAP作用后肝组织抗氧化因子CAT和NRF2 mRNA水平,以及下调炎症因子TNFα、IL-6和CCL5 mRNA水平,且其作用明显优于FGF1^(△HBS)(均P<0.05)。肝素能明显增强FGF1^(△HBS)对抗氧化应激和炎症因子的调节作用(均P<0.05),使其达到与FGF1WT相类似效果。但是,肝素本身没有治疗作用,也不能进一步增强FGF1WT的治疗、抑制和调节作用。结论:肝素参与了FGF1^(WT)对APAP所致小鼠肝脏组织氧化应激、炎症和急性损伤的缓解作用。
Objective:To explore the function and mechanism of wild type heparin in fibroblast growth factor 1(FGF1WT)alleviating acute liver injury(ALI)induced by acetaminophen(APAP).Methods:The experiment was divided into two batches.In the first batch,C57BL/6J mice were randomly divided into five groups including the control group,APAP group,APAP+FGF1WT group,APAP+FGF1WT+Heparin group,APAP+FGF1 mutant(FGF1^(ΔHBS))group and APAP+FGF1^(ΔHBS)+Heparin group.In the second batch,C57BL/6J mice were randomly divided into four groups including the control group,the Heparin group,APAP group and APAP+Heparin group.After 1 h intraperitoneal injection of 500 mg/kg APAP,the corresponding dose of FGF1(2 mg/kg),FGF1^(ΔHBS)(2 mg/kg)and/or heparin(1.5 mg/kg)was intraperitoneally injected.All mice were euthanized 6 h after the above treatment.The damage area of the liver was observed by HE staining.Oxidative stress of the liver was detected by DHE staining.Serum levels of ALT,AST,MDA and SOD in the liver were determined via using commercial kits.Inflammation of the liver was detected by F4/80 immunofluorescence.MRNA expression of CAT,NRF2,CCL5,TNFα and IL-6 were determined by qPCR.All methods were used to explore the therapeutic effects of FGF1^(WT) and FGF1^(ΔHBS) on APAP induced ALI with or without the existence of heparin.Results:The therapeutic effect of FGF1WT on APAP induced ALI was better than FGF1ΔHBS(P<0.05).Heparin could enhance the therapeutic effect of FGF1ΔHBS to a comparable degree of FGF1WT(P<0.05).The inhibition of FGF1^(WT) on APAP induced oxidative stress and inflammation in the liver was stronger than FGF1ΔHBS(P<0.05).Heparin could enhance the inhibitive effect of FGF1^(ΔHBS) to a comparable degree of FGF1WT(P<0.05).The upregulation of mRNA levels of anti-oxidative stress factors including CAT and NRF2 and downregulation of mRNA levels of inflammation factors including TNFα,IL-6 and CCL5 by FGF1^(WT) were stronger than FGF1^(ΔHBS) in the liver of mice treated with APAP(P<0.05).Heparin could enhance the regulatory effect of FGF1ΔHBS on mRNA levels of related anti-oxidative stress factors and inflammation factors to a comparable degree of FGF1^(WT)(P<0.05).However,heparin itself had no therapeutic effect,and cannot further enhance the therapeutic,inhibitory or regulatory effects of FGF1^(WT).Conclusion:Heparin is involved in FGF1WT mediated alleviation of acetaminophen-induced acute liver injury via improving oxidative stress and inflammation.
作者
谢先海
姚瑞娜
范金霞
应赟赟
XIE Xianhai;YAO Ruina;FAN Jinxia;YING Yunyun(Department of Trauma and Emergency,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China;School of Basic Medical Sciences,Wenzhou Medical University,Wenzhou 325035,China;Department of Gastrointestinal Surgery,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,China)
出处
《温州医科大学学报》
2025年第10期797-804,共8页
Journal of Wenzhou Medical University
基金
温州市基础性科研项目(Y20210694)。
