刺五加多糖调控炎性因子对小鼠免疫性肝损伤的保护作用
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摘要
探讨刺五加多糖(ASPs)对刀豆蛋白A(Con A)造成的小鼠免疫性肝损伤模型的保护作用和机制。将BALB/c小鼠随机分成7组,对照组,模型组,ASPs低、中、高剂量组(36. 25,72. 5,145 mg·kg~(-1)),联苯双酯滴丸(阳性药,200 mg·kg~(-1))组,吡咯烷二硫代氨基甲酸盐(PDTC,NF-κB抑制剂,200 mg·kg~(-1))组。ASPs,联苯双酯滴丸组每天灌胃给药1次,连续7 d;对照组、模型组和PDTC组每天以10 m L·kg~(-1)生理盐水灌胃给药。于末次灌胃后,PDTC组灌胃PDTC,再于0. 5 h后,除对照组外均尾静脉注射20 mg·kg~(-1)Con A诱导肝损伤。8 h后处死动物取材,对肝脏组织进行组织病理学检查,试剂盒法检测肝组织匀浆中一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MDA)、还原性谷胱甘肽(GSH-PX)、白细胞介素-1β(IL-1β)及肿瘤坏死因子-α(TNF-α)水平,采用Western blot法检测肝组织中TNF-α,ICAM-1,i NOS和NF-κB蛋白的表达水平,探讨刺五加多糖对治疗免疫性肝损伤对肝的保护作用和机制。与模型组比较,炎性细胞浸润减轻,肝细胞胞浆疏松肿胀、核固缩浓染得以改善,ASPs显著降低肝组织匀浆中MDA,NO,IL-1β,TNF-α的含量;升高GSH-PX和SOD的含量;降低肝组织中TNF-α,ICAM-1,i NOS和NF-κB蛋白水平表达。ASPs降低炎性细胞因子和黏附因子的活性、减少炎性细胞因子的分泌和表达,对免疫性肝损伤有一定保护作用。
The aim of this paper was to discuss the protective effect and mechanism of Acanthopanax senticosus polysaccharides( ASPs) on immunological liver injury caused by conanavalin A( Con A). BALB/c mice were randomly divided into seven groups: control group,model group( Con A),low-,medium-,and high-dose( 36. 25,72. 5,145 mg·kg~(-1)) ASPs groups,bifendate( 200 mg·kg~(-1),positive drug) group and pyrrolidinedithiocarbamate( PDTC,NF-κB inhibitor,200 mg·kg~(-1)) group. ASPs groups and bifendate group were given with corresponding drugs by ig administration once daily for 7 d. Control group,model group and PDTC group were given with normal saline by ig administration once daily for 7 d. After the last ig administration,PDTC was given in DTC group by iv administration( 200 mg·kg~(-1)); 0. 5 h after that,Con A( 20 mg·kg~(-1)) was injected via the tail vein to induce immunological liver injury in all the mice except normal control group. The mice were killed 8 h later and their liver tissues were collected for histopathological examination. The contents of nitric oxide( NO),superoxide dismutase( SOD),malondialdehyde( MDA),reduced glutathione( GSHPX),interleukin( IL-1β) and tumor necrosis factor( TNF-α) in liver tissues were detected by kit assay. Western blot method was used to detect TNF-α,intercellular cell adhesion molecule-1( ICAM-1),inducible nitric oxide synthase( i NOS) and nuclear factor( NF-κB) protein expression in liver tissues. As compared with model group,ASPs not only could reduce the activity of MDA,NO,IL-1β and TNF-α,but also increase the content of GSH-PX and SOD; at the same time,the protein expression levels of TNF-α,ICAM-1,i NOS and NF-κB were reduced in liver tissues; in addition,inflammatory cell infiltration was alleviated,hepatocyte cytoplasm was loose and swollen,and nuclear condensation and staining were improved. ASPs has a protective effect on immunological liver injury,and the mechanism may be associated with regulating secretion of inflammatory cytokines and the expression of adhesion factor through NF-κB signaling pathway.
The aim of this paper was to discuss the protective effect and mechanism of Acanthopanax senticosus polysaccharides( ASPs) on immunological liver injury caused by conanavalin A( Con A). BALB/c mice were randomly divided into seven groups: control group,model group( Con A),low-,medium-,and high-dose( 36. 25,72. 5,145 mg·kg~(-1)) ASPs groups,bifendate( 200 mg·kg~(-1),positive drug) group and pyrrolidinedithiocarbamate( PDTC,NF-κB inhibitor,200 mg·kg~(-1)) group. ASPs groups and bifendate group were given with corresponding drugs by ig administration once daily for 7 d. Control group,model group and PDTC group were given with normal saline by ig administration once daily for 7 d. After the last ig administration,PDTC was given in DTC group by iv administration( 200 mg·kg~(-1)); 0. 5 h after that,Con A( 20 mg·kg~(-1)) was injected via the tail vein to induce immunological liver injury in all the mice except normal control group. The mice were killed 8 h later and their liver tissues were collected for histopathological examination. The contents of nitric oxide( NO),superoxide dismutase( SOD),malondialdehyde( MDA),reduced glutathione( GSHPX),interleukin( IL-1β) and tumor necrosis factor( TNF-α) in liver tissues were detected by kit assay. Western blot method was used to detect TNF-α,intercellular cell adhesion molecule-1( ICAM-1),inducible nitric oxide synthase( i NOS) and nuclear factor( NF-κB) protein expression in liver tissues. As compared with model group,ASPs not only could reduce the activity of MDA,NO,IL-1β and TNF-α,but also increase the content of GSH-PX and SOD; at the same time,the protein expression levels of TNF-α,ICAM-1,i NOS and NF-κB were reduced in liver tissues; in addition,inflammatory cell infiltration was alleviated,hepatocyte cytoplasm was loose and swollen,and nuclear condensation and staining were improved. ASPs has a protective effect on immunological liver injury,and the mechanism may be associated with regulating secretion of inflammatory cytokines and the expression of adhesion factor through NF-κB signaling pathway.
引文
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[14] Yu H B,Zhang H F,Zhang X,et al. Resveratrol inhibits VEGF expression of human hepatocellular carcinoma cells through a NF-kappa B-mediated mechanism[J]. Hepatogastroenterology,2010,57(102/103):1241.
[15]罗景慧,杨迎暴.白黎芦醇对酒精性肝损伤大鼠血清中TNFa、IL-6、ICAM-1水平的影响及机制[J].中药药理与临床,2010,26(4):14.
[16]龙爽,刘绍勇,徐英.痰热清注射液及5种中间体对四氯化碳致急性肝损伤大鼠的保护作用探讨[J].中国实验方剂学杂志,2018,24(11):73.
[17] Kuboki S,Okaya T,Schuster R,et al. Hepatocyte NF-kappaB activation is hepatoprotective during ischemia-reperfusion injury and is augmented by ischemic hypothermia[J]. Am J physiol Gastrointest Liver Physiol,2007,292(1):201.
[2] Mahi H,Gores G J. Cellular and molecular mechanisms of liver injury[J]. J Pathol,2008,134(6):1641.
[3]袁媛,吴芹,石京山,等.丹参及其主要成分保肝作用得研究进展[J].中国中药杂志,2015,40(4):588.
[4]刘树民,张娜.刺五加多糖的现代研究进展[J].中医药信息,2014,31(2):116.
[5] Wang H X,Liu M,Weng S Y,et al. Immune mechanisms of concanavalin A model of autoimmune hepatitis[J]. World J Gastroenterol,2012,18(2):119.
[6] Chen M,Cao L,Luo Y,et al. Paeoniflorin protects against concanavalin A-induced hepatitis in mice[J]. Int Immunopharmacol,2015,24(1):42.
[7]卢芳,杨晓丹,张颖,等.基于代谢组学方法探讨刺五加多糖对免疫性肝损伤小鼠的保护作用[J].中药新药与临床药理,2016,27(6):823.
[8]张娜,毛迪,安柏松.刺五加多糖对免疫性肝损伤BALB/c小鼠IL-2、IL-2、INF-γ细胞因子及mRNA表达的影响[J].药物评价研究,2018,41(4):557.
[9] Wang K P,Song Z Z,Wang H J,et al. Angelica sinensis polysaccharide attenuates concanavalin A-induced liver injury in mice[J].Int Immunopharmacol,2016,31(2):140.
[10] Tan Y W,Wu J C. Establishment of rats model with acute hepatic injury induced by injection of concanavalin A[J]. Jiangsu Med J,2009,35(6):702.
[11]曲建慧,张修礼.ConA性小鼠肝损伤模型的研究及应用[J].肝脏,2001,6(3):180.
[12]刘悦晖,范学工,李宁,等.刀豆球蛋白A所致实验性肝损伤模型的构建[J].中国感染控制杂志,2008,7(5):297.
[13] Kobayashi S,Nishihira J,Watanabe S,et al. Prevention of lethal acute hepatic failure by antimacrophage migration inhibitory factory antibody in mice treated with bacille calmette-guerin and lipopolysaccharide[J]. Hepatology,1999,29(6):1752.
[14] Yu H B,Zhang H F,Zhang X,et al. Resveratrol inhibits VEGF expression of human hepatocellular carcinoma cells through a NF-kappa B-mediated mechanism[J]. Hepatogastroenterology,2010,57(102/103):1241.
[15]罗景慧,杨迎暴.白黎芦醇对酒精性肝损伤大鼠血清中TNFa、IL-6、ICAM-1水平的影响及机制[J].中药药理与临床,2010,26(4):14.
[16]龙爽,刘绍勇,徐英.痰热清注射液及5种中间体对四氯化碳致急性肝损伤大鼠的保护作用探讨[J].中国实验方剂学杂志,2018,24(11):73.
[17] Kuboki S,Okaya T,Schuster R,et al. Hepatocyte NF-kappaB activation is hepatoprotective during ischemia-reperfusion injury and is augmented by ischemic hypothermia[J]. Am J physiol Gastrointest Liver Physiol,2007,292(1):201.
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