ROS介导IL-17A诱导的TF表达或有助于治疗TF相关的肝硬化
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摘要
为了评估白细胞介素IL-17A对肝硬化组织中内皮细胞(endothelial cells, EC)组织因子(tissue factor, TF)表达的调节作用,本研究选择50例肝硬化患者(肝硬化组)和9例创伤性脾破裂患者(对照组)作为研究对象,通过检测两组脾静脉内皮上的TF表达和血清IL-17A水平,以及IL-17A诱导后人脐静脉内皮细胞(human umbilical vein endothelial cell, HUVEC)中的TF表达和活性。另外,考察活性氧簇(reactive oxygen species, ROS)对肝硬化患者血清中内皮细胞TF表达和活性的影响。研究显示,肝硬化组的脾静脉内皮上的TF表达显著高于对照组,与对照组(24.65±4.02) pg/m L相比,肝硬化组IL-17A水平显著升高(46.78±7.68) pg/mL。用不同浓度和时间的IL-17A诱导HUVEC,可导致TF表达和活性呈浓度和时间依赖性升高。IL-17A (100 ng/mL)处理可诱导HUVEC中活性氧簇(reactive oxygen species, ROS)水平逐渐升高,6 h以后逐渐降低。使用10 mmol/L NAC (ROS清除剂)和2 mg/mL IL-17受体C抗体(Interleukin-17 receptor A, anti-IL-17RA)预处理HUVEC 2h,可抑制TF表达和活性。添加人IL-17抗体(1μg/mL)和NAC (10 mmol/L)的两组血清TF表达和活性显著降低。另外,对照组血清中IL-17A的添加量达到80 pg/mL时,TF活性及表达水平与肝硬化组一致。本研究初步结论表明IL-17A诱导的TF表达通过活化ROS介导,靶向IL-17A和ROS信号通路可能有助于治疗与TF相关的肝硬化。
In order to evaluate the regulation of IL-17 A on endothelial cell(EC) tissue factor(TF) expression in cirrhotic tissues, in this study, 50 patients with cirrhosis(liver cirrhosis group) and 9 patients with traumatic spleen rupture(control group) were selected as subjects. The TF expression and serum IL-17 A levels in the spleen veins of the two groups were tested, as well as TF expression and activity in human umbilical vein endothelial cells(HUVEC) after IL-17 A induction. In addition, the effect of reactive oxygen species(ROS) on the expression and activity of TF in endothelial cells in serum of patients with liver cirrhosis was investigated. Studies have shown that the expression of TF on the spleen vein endothelium in the cirrhosis group was significantly higher than that in the control group. Compared with the control group(24.65±4.02) pg/mL, the level of IL-17 A in the cirrhosis group significantly increased(46.78±7.68) pg/mL. Stimulation of HUVEC with different concentrations and time duration of IL-17 A resulted in a concentration and time-dependent increase in TF expression and activity.Treatment with IL-17 A(100 ng/mL) induced a gradual increase in ROS levels in HUVECs and decreased gradually after 6 h. Pretreatment of HUVECs with 10 mmol/L NAC(ROS scavenger) and 2 mg/mL anti-IL-17 RA(IL-17 receptor antibody) for 2 h inhibited TF expression and activity. The TF expression and activity significantly reduced in the two groups of serum supplemented with human IL-17 antibody(1 μg/mL) and NAC(10 mmol/L).In addition, when the amount of IL-17 A addition in the serum of the control group reached 80 pg/mL, the TF activity and expression level were consistent with that of the cirrhosis group. The preliminary conclusions of this study demonstrated that IL-17 A-induced TF expression would be mediated through activation of ROS, and targeting IL-17 A and ROS signaling pathways might help to treat cirrhosis associated with TF.
In order to evaluate the regulation of IL-17 A on endothelial cell(EC) tissue factor(TF) expression in cirrhotic tissues, in this study, 50 patients with cirrhosis(liver cirrhosis group) and 9 patients with traumatic spleen rupture(control group) were selected as subjects. The TF expression and serum IL-17 A levels in the spleen veins of the two groups were tested, as well as TF expression and activity in human umbilical vein endothelial cells(HUVEC) after IL-17 A induction. In addition, the effect of reactive oxygen species(ROS) on the expression and activity of TF in endothelial cells in serum of patients with liver cirrhosis was investigated. Studies have shown that the expression of TF on the spleen vein endothelium in the cirrhosis group was significantly higher than that in the control group. Compared with the control group(24.65±4.02) pg/mL, the level of IL-17 A in the cirrhosis group significantly increased(46.78±7.68) pg/mL. Stimulation of HUVEC with different concentrations and time duration of IL-17 A resulted in a concentration and time-dependent increase in TF expression and activity.Treatment with IL-17 A(100 ng/mL) induced a gradual increase in ROS levels in HUVECs and decreased gradually after 6 h. Pretreatment of HUVECs with 10 mmol/L NAC(ROS scavenger) and 2 mg/mL anti-IL-17 RA(IL-17 receptor antibody) for 2 h inhibited TF expression and activity. The TF expression and activity significantly reduced in the two groups of serum supplemented with human IL-17 antibody(1 μg/mL) and NAC(10 mmol/L).In addition, when the amount of IL-17 A addition in the serum of the control group reached 80 pg/mL, the TF activity and expression level were consistent with that of the cirrhosis group. The preliminary conclusions of this study demonstrated that IL-17 A-induced TF expression would be mediated through activation of ROS, and targeting IL-17 A and ROS signaling pathways might help to treat cirrhosis associated with TF.
引文
A nsari S.A.,Pendurthi U.R.,and Rao L.V.M.,2017,The lipid peroxidation product 4-hydroxy-2-nonenal induces tissue factor decryption via ROS generation and the thioredoxin system,Blood Adv.,1(25):2399-2413
Devaraj S.,Dasu M.U.,Rao L.,and Jialal I.,2009,C-reactive protein stimulates superoxide anion release and tissue factor activity in vivo,Atherosclerosis,203(1):67-74
Du W.J.,Zhen J.H.,Zeng Z.Q.,Zheng Z.M.,Xu Y.,Qin L.Y.,and Chen S.J.,2013,Expression of Interleukin-17 associated with disease progression and;liver fibrosis with hepatitis B virus infection:IL-17 in HBV infection,Diagnostic Pathology,8(1):40
Hatakeyama K.,Asada Y.,Marutsuka K.,Kataoka H.,Sato Y.,and Sumiyoshi A.,1998,Expression of tissue factor in the rabbit aorta after balloon injury,Atherosclerosis,139(2):265-271
Lv B.,Wang H.,Tang Y.,Fan Z.,Xiao X.,and Chen F.,2009,High-mobility group box 1 protein induces tissue factor expression in vascular endothelial cells via activation of NF-kappaB and Egr-1,Thrombosis&Haemostasis,101(2):352-359
Miyoshi T.,Yamashita K.,Arai T.,Yamamoto K.,Mizugishi K.,and Uchiyama T.,2010,The role of endothelial interleukin-8/NADPH oxidase 1 axis in sepsis,Immunology,131(3):331-339
Peng Y.,Qi X.,and Guo X.,2016,Child-Pugh versus MELDscore for the assessment of prognosis in liver cirrhosis:a systematic review and meta-analysis of observational stud ies,Medicine,95(8):e2877
Penn M.S.,and Topol E.J.,2001,Tissue factor,the emerging link between inflammation,thrombosis,and vascular remodeling,Circulation Research,89(1):1-2
Pietrowski E.,Bender B.,Huppert J.,White R.,Luhmann H.J.,and Kuhlmann C.R.,2011,Pro-inflammatory effects of interleukin-17A on vascular smooth muscle cells involve NAD(P)H-oxidase derived reactive oxygen species,Jour nal of Vascular Research,48(1):52-58
Shen Q.,Goderie S.K.,Jin L.,Karanth N.,Sun Y.,Abramova N.,Vincent P.,Pumiglia K.,and Temple S.,2004,Endothelial cells stimulate self-renewal and expand neurogenesis of neural stem cells,Science,304(5675):1338-1340
Versteeg H.H.,Peppelenbosch M.P.,and Spek C.A.,2001,The pleiotropic effects of tissue factor:a possible role for factor VIIa-induced intracellular signalling?Thromb Haemost,86(6):1353-1359
Devaraj S.,Dasu M.U.,Rao L.,and Jialal I.,2009,C-reactive protein stimulates superoxide anion release and tissue factor activity in vivo,Atherosclerosis,203(1):67-74
Du W.J.,Zhen J.H.,Zeng Z.Q.,Zheng Z.M.,Xu Y.,Qin L.Y.,and Chen S.J.,2013,Expression of Interleukin-17 associated with disease progression and;liver fibrosis with hepatitis B virus infection:IL-17 in HBV infection,Diagnostic Pathology,8(1):40
Hatakeyama K.,Asada Y.,Marutsuka K.,Kataoka H.,Sato Y.,and Sumiyoshi A.,1998,Expression of tissue factor in the rabbit aorta after balloon injury,Atherosclerosis,139(2):265-271
Lv B.,Wang H.,Tang Y.,Fan Z.,Xiao X.,and Chen F.,2009,High-mobility group box 1 protein induces tissue factor expression in vascular endothelial cells via activation of NF-kappaB and Egr-1,Thrombosis&Haemostasis,101(2):352-359
Miyoshi T.,Yamashita K.,Arai T.,Yamamoto K.,Mizugishi K.,and Uchiyama T.,2010,The role of endothelial interleukin-8/NADPH oxidase 1 axis in sepsis,Immunology,131(3):331-339
Peng Y.,Qi X.,and Guo X.,2016,Child-Pugh versus MELDscore for the assessment of prognosis in liver cirrhosis:a systematic review and meta-analysis of observational stud ies,Medicine,95(8):e2877
Penn M.S.,and Topol E.J.,2001,Tissue factor,the emerging link between inflammation,thrombosis,and vascular remodeling,Circulation Research,89(1):1-2
Pietrowski E.,Bender B.,Huppert J.,White R.,Luhmann H.J.,and Kuhlmann C.R.,2011,Pro-inflammatory effects of interleukin-17A on vascular smooth muscle cells involve NAD(P)H-oxidase derived reactive oxygen species,Jour nal of Vascular Research,48(1):52-58
Shen Q.,Goderie S.K.,Jin L.,Karanth N.,Sun Y.,Abramova N.,Vincent P.,Pumiglia K.,and Temple S.,2004,Endothelial cells stimulate self-renewal and expand neurogenesis of neural stem cells,Science,304(5675):1338-1340
Versteeg H.H.,Peppelenbosch M.P.,and Spek C.A.,2001,The pleiotropic effects of tissue factor:a possible role for factor VIIa-induced intracellular signalling?Thromb Haemost,86(6):1353-1359