灯盏花素对肝脏缺血再灌注损伤大鼠肝脏和肠黏膜屏障功能的保护作用*

doi:10.3969/j.issn.1672-5069.2019.05.006

摘要/Abstract

摘要： 目的探讨灯盏花素对肝脏缺血再灌注损伤（IRI）大鼠肝脏和肠黏膜屏障功能的保护作用。方法随机将45只SD大鼠分为对照组、模型组和灯盏花素干预组,制备肝脏缺血再灌注损伤和肠缺血再灌注损伤模型,采用ELISA法检测肠黏膜分泌型（S）IgA和血浆内毒素水平,检测肠组织诱导型一氧化氮合成酶（iNOS）、内皮型一氧化氮合成酶（eNOS）和一氧化氮（NO）水平,使用试剂盒检测血浆降钙素原（ PCT）、二胺氧化酶（DAO）、TNF-α和IL-1β,采用Western Blotting法检测肝组织Toll样受体-4（TLR4）和核因子（NF）-κB表达水平。结果模型组大鼠肠粘膜SlgA水平为（0.2±0.1）μg/kg,显著低于对照组【（0.7±0.1）μg/kg,P<0.01】,血清内毒素水平为（0.8±0.1）EU/ml,显著高于对照组【（0.2±0.1） EU/ml,P<0.01】,肠组织iNOS为（0.7±0.1） U/g,显著高于对照组【（0.2±0.1） U/g,P<0.01】,而eNOS为（0.2±0.1） U/g,显著低于对照组【（0.4±0.1）U/g,P<0.01】, NO为（0.2±0.1）μmol/g,显著低于对照组【（0.4±0.0）μmol/g,P<0.01】, PCT为（5.0±1.2） ng/ml,显著高于对照组【（3.5±0.98）ng/ml,P<0.01】,DAO为（10.5±1.6） Ku/l,显著高于对照组【（6.5±1.2） Ku/l,P<0.01】;灯盏花素干预组SlgA为（0.7±0.1）μg/kg,显著高于模型组【（0.2±0.1）μg/kg,P<0.01】,内毒素为（0.3±0.1） EU/ml,显著低于模型组【（0.8±0.1） EU/ml,P<0.01】,iNOS 为（0.3±0.1） U/g,显著低于模型组【（0.7±0.1） U/g,P<0.01】, eNOS为（0.8±0.2） U/g,显著高于模型组【（0.2±0.1）U/g,P<0.01】,NO为（0.8±0.1）μmol/g,显著高于模型组【（0.2±0.1）μmol/g,P<0.01】,PCT为（3.9±1.0） ng/ml,显著低于模型组【（5.0±1.2） ng/ml,P<0.01】,DAO为（7.5±1.3） Ku/L,显著低于模型组【（10.5±1.6） Ku/l,P<0.01】;模型组大鼠肝组织TLR4和NF-κB表达及血清TNF-α和IL-1β水平显著高于对照组（P<0.01）,而灯盏花素干预组大鼠肝组织TLR4和NF-κB及血清TNF-α和IL-1β水平显著降低,差异有统计学意义（P<0.01）。结论灯盏花素通过促进eNOS／NO表达和下调TLR4和NF-κB表达能降低血浆内毒素对肝脏的损伤,对IRI动物具有保护作用。

关键词: 缺血再灌注损伤, 肝脏, 灯盏花素, 肠黏膜, 大鼠

Abstract: Objective The aim of this study was to explore the protective effects of erigeron breviscapus,a herbal medicine,on liver and intestinal mucosal barrier functions in rats with hepatic ischemia-reperfusion injury （IRI）. Methods 45 SD rats were randomly divided into control,model and breviscapine-intervened group,and the hepatic and intestinal IRI models were established. The intestinal mucosal SIgA and plasma endotoxin were detected by ELISA,and the inducible nitric oxide synthase （iNOS）,endothelial （eNOS） and total nitric oxide （NO） in intestinal tissues were detected. Serum procalcitonin （PCT）,diamine oxidase （DAO）,TNF-α and IL-1β levels were detected by kits. The Toll-like receptor 4（TLR4） and nuclear factor （NF）-κB were detected by Western blotting. Results The SlgA in model group was significantly lower than that in control group [（0.2±0.1） μg/kg vs. （0.7±0.1） μg/kg,P<0.01],the endotoxin in model group was significantly higher than that in control [（0.8±0.1） EU/ml vs.（0.2±0.1） EU/ml,P<0.01],the iNOS in model group was significantly higher than that in control group [（0.7±0.1）U/g vs.（0.2±0.1） U/g,P<0.01],the eNOS in model group was significantly lower than that in control group [（0.2±0.1） U/g vs. （0.4±0.1） U/g,P<0.01],and the NO in model group was significantly lower than that in control group [（0.2±0.1） μmol/g vs. （0.4±0.0） μmol/g,P<0.01];the PCT in breviscapine-intervened group was significantly higher than that in model group [（5.0±1.2） ng/ml vs.（3.5±0.98） ng/ml,P<0.01],the SlgA in breviscapine intervention group was significantly higher than that in model group [（0.7±0.1） μg/kg vs.（0.2±0.1）μg/kg,P<0.01],while the endotoxin was significantly lower than that in model group [（0.3±0.1） EU/ml vs.（0.8±0.1） EU/ml,P<0.01],the iNOS was significantly lower than that in model group [（0.3±0.1） U/g vs.（0.7±0.1 U/g,P<0.01],the eNOS was significantly higher than that in model group [（0.8±0.2） U/g vs. （0.2±0.1） U/g,P<0.01],and the NO was significantly higher than that in model group[（0.8±0.1） μmol/g vs.（0.2±0.1） μmol/g,P<0.01];the PCT was significantly lower than that in the model group [（3.9±1.0） ng/ml vs. （5.0±1.2） ng/ml,P<0.01],and the DAO was significantly lower than that in the model group [（7.5±1.3） Ku/L vs.（10.5±1.6） Ku/L,P<0.01];the expression of TLR4 and NF-κB in liver tissues and serum TNF-α and IL-1β levels in model group were significantly higher than those in control group （P<0.01）,while they all significantly decreased in breviscapine-intervened group （P<0.01）. Conclusion s Breviscapine might protect hepatic and intestinal mucosal barrier functions in rats with IRI by promoting eNOS/NO expression and down-regulating expression of TLR4 and NF-κB, which might reduce plasma endotoxin.

Key words: Ischemia-reperfusion injury, Liver, Intestinal mucosa, Erigeron breviscapus, Rats

林秋香, 王雪雯, 黄祖雄, 潘晨. 灯盏花素对肝脏缺血再灌注损伤大鼠肝脏和肠黏膜屏障功能的保护作用*[J]. 实用肝脏病杂志, 2019, 22(5): 636-639.

Lin Qiuxiang, Wang Xuewen, Huang Zuxiong, et al. Protective effects of breviscapine on liver and intestinal mucosal barrier functions in rats with hepatic ischemia-reperfusion injury[J]. Journal of Practical Hepatology, 2019, 22(5): 636-639.

参考文献

[1] Wijck KV,Buurman WA.Ischemia-reperfusion injury. J Surg Res,2002,105(2):248-258.
[2] Kadono K,Uchida Y,Hirao H,et al.Thrombomodulin attenuates inflammatory damage due to liver ischemia and reperfusion injury in mice in TLR4-dependent manner.Am J Transplant,2016,17(1):69-80.
[3] Tong Y,Ding X B,Chen Z X,et al.WISP1 mediates hepatic warm ischemia reperfusion injury via TLR4 signaling in mice. Sci Rep,2016,6(29):20141-20143.
[4] Lin Y Z,Lu Z Y,Liang X H,et al.Effect of breviscapine against hepatic ischemia reperfusion injury. J Surg Res,2016,203(2):268-274.
[5] Bao Z,Chen W,Pan F,et al.Role of mitofusin 2 in the protective effect of breviscapine against hepatic ischemia/reperfusion injury in rats. Exp Ther Med,2018,15(4):3582-3588.
[6] Pengyue Z,Tao G,Hongyun H,et al.Breviscapine confers a neuroprotective efficacy against transient focal cerebral ischemia by attenuating neuronal and astrocytic autophagy in the penumbra. Biomed Pharmacother,2017,90:69-76.
[7] Zhang B,Liu Q H,Zhou C J,et al.Protective effect of eNOS overexpression against ischemia/reperfusion injury in small-for-size liver transplantation. Exp Ther Med,2016,12(5):3181-3188.
[8] Turner J R.Intestinal mucosal barrier function in health and disease. Nat Rev Immunol,2009,9(11):799-809.
[9] Ferchichi H,Bacha S,Kourda N,et al.Animal model of liver ischemia reperfusion:biochemical and histological evaluation. Tunis Med,2016,94(3):235.
[10] Aggarwal B B.Nuclear factor-κB. Can Cell,2004,1(1):11-13.
[11] Xiong R.Effect of ecological immune enteral nutrition intervention on intestinal barrier function and systemic inflammatory response in rat models with severe pancreatitis. J Hain Medl Uni,2016,22(3):12-16.
[12] Couto D,Zipfel C.Regulation of pattern recognition receptor signalling in plants. Nat Rev Immunol,2016,16(9):537-552.
[13] Wang J,Ji S Y,Liu S Z,et al.Cardioprotective effect of breviscapine:inhibition of apoptosis in H9c2 cardiomyocytes via the PI3K/Akt/eNOS pathway following simulated ischemia/reperfusion injury. Pharmazie,2015,70(9):593-597.
[14] Riquelme J A,Westermeier F,Hall A R,et al.Dexmedetomidine protects the heart against ischemia-reperfusion injury by an endothelial eNOS/NO dependent mechanism. Pharmacol Res,2016,103(1):318-327.
[15] Xiao H B,Sui G G,Lu X Y.Icariin improves eNOS/ NO-pathway to prohibit the atherogenesis of apolipoprotein E null mice. Can J Physiol Pharmacol, 2016,95(6):625-633.
[16] Li H C,Fan X J,Chen Y F,et al.Early prediction of intestinal mucosal barrier function impairment by elevated serum procalcitonin in rats with severe acute pancreatitis. Pancreatology,2016,16(2):211-217.
[17] Ding Y,Li Y,Zhang X,et al.Soluble epoxide hydrolase activation by S-nitrosation contributes to cardiac ischemia-reperfusion injury. J Mol Cell Cardiol,2017,110:70-79.
[18] Li H C,Fan X J,Chen Y F,et al.Early prediction of intestinal mucosal barrier function impairment by elevated serum procalcitonin in rats with severe acute pancreatitis. Pancreatology,2016,16(2):211-217.
[19] Jie H,Xu Y,Di Y,et al.Effect of polysaccharides from acanthopanax senticosuson intestinal mucosal barrier of Escherichia coli lipopolysaccharide challenged mice. Asian Australas J Anim Sci,2016,29(1):134-141.
[20] Ju H,Li H,Tian T,et al.Melatonin overcomes gemcitabine resistance in pancreatic ductal adenocarcinoma by abrogating nuclear factor-κB activation.J Pineal Res,2016,60(1):27-38.