肠愈灌肠方对溃疡性结肠炎大鼠炎症相关因子及DOR、β-arrestin1、Bcl-2表达影响
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摘要
目的:探讨肠愈灌肠方对溃疡性结肠炎(Ulcerative Colitis,UC)大鼠肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)及δ阿片受体(DOR)、β-抑制蛋白1(β-arrestin1)、Bcl-2表达的影响。方法:SD大鼠60只,随机分为正常组、模型组、肠愈灌肠方组和美沙拉嗪组,除正常组外,其余大鼠采用5%TNBS灌肠建立大鼠溃疡性结肠炎模型。肠愈灌肠方组、美沙拉嗪组分别予肠愈灌肠方药液和美沙拉嗪悬浊液灌肠,正常组和模型组给予蒸馏水灌肠,连续15d,第16d处死大鼠,取结肠标本。分别采用Elisa和Western Blotting法检测结肠组织中TNF-α、T GF-β1及DOR、β-arrestin1、Bcl-2蛋白的表达。结果:模型组大鼠体重下降、精神萎靡,出现不同程度的腹泻及便血,造模3d后DAI评分高于正常组(P<0.01),大鼠结肠肠管变粗、肠壁增厚,黏膜水肿、充血。肠愈灌肠方能明显改善模型大鼠的结肠炎症损伤,与正常组比较,模型组的大鼠结肠黏膜组织抑炎因子TGF-β1显著降低(P<0.01),炎症因子TNF-α及DOR、β-arrestin1、Bcl-2表达水平显著增高(P<0.01);与模型组比较,肠愈灌肠方组与美沙拉嗪组抑炎因子TGFβ1明显升高(P<0.01),炎症因子TNF-α及DOR、β-arrestin1、Bcl-2蛋白表达明显下降(P<0.01,P<0.05),但肠愈灌肠方组和美沙拉嗪组之间比较无显著差异。结论:在实验性溃疡性结肠炎大鼠结肠组织中抑炎因子TGF-β1减少,炎症因子TNF-α及DOR、β-arrestin1、Bcl-2的表达升高,炎症相关因子及DOR-β-arrestin1-Bcl-2信号转导通路可能参与了溃疡性结肠炎的发展过程,肠愈灌肠方对其改善作用的机制可能与调节该信号通路有关。
Objective: To explore the effect of Changyu Guanchang recipe on the expression of inflammation related factors TNF-α, TGFβ1 and DOR、β-arrestin1、Bcl-2 in rats with ulcerative colitis.Methods 60 SD rats were randomly divided into normal group, model group,Changyu Guanchang recipe group and Mesalazine group.In addition to the normal group, the rest rats were used to establish the model of ulcerative colitis in rats by 5%TNBS enema. Changyu Guanchang recipe group and Mesalazine group were given the Changyu Guanchang recipe and Mesalazine, normal group and model group were given distilled water enema for 15 days, sixteenth days, rats were killed, and the colon specimens were taken. Elisa and Western Blotting respectively were used to detect the expression of TNF-α, TGFβ1 and DOR、β-arrestin1、Bcl-2 protein in colon tissues. Results Body weight of rats in model group decreased, listlessness, diarrhea and hematochezia in different degree, DAI score higher than the normal group 3 days after modeling(P<0.01).The rat colon intestinal thicker wall thickening, mucosal edema, hyperemia. Changyu Guanchang recipe can significantly improve the rat model of colitis in rats.Compared with the normal group, the model group of rat colonic mucosal tissue anti-inflammatory factor TGF-β1 significantly decreased(P<0.01), the levels of inflammatory factors TNF-α,and DOR、β-arrestin1、Bcl-2 expression was significantly increased(P<0.01); Compared with model group, the expression of TGF-β was significantly increased in Changyu Guanchang recipe group and Mesalazine group(P<0.01),the expression of TNF-α and DOR、β-arrestin1、Bcl-2 were significantly decreased(P<0.01, P<0.05). There was no significant difference between the Changyu Guanchang recipe group and Mesalazine group. Conclusion The signal transduction pathway of inflammatory cytokines and DOR-β-arrestin1-Bcl-2 may be involved in the development of ulcerative colitis. The mechanism of the effect of the intestinal healing enema may be related to the regulation of the signaling pathway.
Objective: To explore the effect of Changyu Guanchang recipe on the expression of inflammation related factors TNF-α, TGFβ1 and DOR、β-arrestin1、Bcl-2 in rats with ulcerative colitis.Methods 60 SD rats were randomly divided into normal group, model group,Changyu Guanchang recipe group and Mesalazine group.In addition to the normal group, the rest rats were used to establish the model of ulcerative colitis in rats by 5%TNBS enema. Changyu Guanchang recipe group and Mesalazine group were given the Changyu Guanchang recipe and Mesalazine, normal group and model group were given distilled water enema for 15 days, sixteenth days, rats were killed, and the colon specimens were taken. Elisa and Western Blotting respectively were used to detect the expression of TNF-α, TGFβ1 and DOR、β-arrestin1、Bcl-2 protein in colon tissues. Results Body weight of rats in model group decreased, listlessness, diarrhea and hematochezia in different degree, DAI score higher than the normal group 3 days after modeling(P<0.01).The rat colon intestinal thicker wall thickening, mucosal edema, hyperemia. Changyu Guanchang recipe can significantly improve the rat model of colitis in rats.Compared with the normal group, the model group of rat colonic mucosal tissue anti-inflammatory factor TGF-β1 significantly decreased(P<0.01), the levels of inflammatory factors TNF-α,and DOR、β-arrestin1、Bcl-2 expression was significantly increased(P<0.01); Compared with model group, the expression of TGF-β was significantly increased in Changyu Guanchang recipe group and Mesalazine group(P<0.01),the expression of TNF-α and DOR、β-arrestin1、Bcl-2 were significantly decreased(P<0.01, P<0.05). There was no significant difference between the Changyu Guanchang recipe group and Mesalazine group. Conclusion The signal transduction pathway of inflammatory cytokines and DOR-β-arrestin1-Bcl-2 may be involved in the development of ulcerative colitis. The mechanism of the effect of the intestinal healing enema may be related to the regulation of the signaling pathway.
引文
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[12]李斌,谷松.乌梅丸及其拆方对溃疡性结肠炎大鼠结肠黏膜组织IL-10与TNF-α的影响[J].辽宁中医药大学学报,2015,17(6):20-22.
[13]王玉芳,魏兵,欧阳钦.转化生长因子β1在溃疡性结肠炎中的表达[J].四川大学学报(医学版),2005,36(2):204-206.
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[15]刘健,杨小玉,董健,等.Bcl-2蛋白与细胞凋亡的关系及其在中枢神经损伤修复中作用的研究进展[J].吉林大学学报(医学版),2012,38(3):603-606.
[16]Kang J,Shi Y,Xiang B,et al.A nuclear function of beta-arrestin1 in GPCR signaling:regulation of histone acetylation and gene transcription[J].Cel,2005,123(5):833-847.
[17]Martinou JC,Youle RJ.Mitochondria in apoptosis:Bcl-2 family members and mitochondrial dynamics[J].Dev Cell,2011,21(1):92-101.
[18]孙仁山,陈晓红,程天民,等.白芨对大鼠创面愈合几个要素的影响[J].中国临床康复,2003,7(29):3927-3929.
[19]何英,李春明,赵巍,等.锡类散治疗溃疡性结肠炎疗效观察[J].中国临床医学,2011,18(3):347-348.
[2]Gyires K,TóthéV,Zádori SZ.Gut inflammation:current update on pathophysiology,molecular mechanism and pharmacological treatment modalities[J].Curr Pharm Des,2014,20(7):1063-81.
[3]Mudter J,Neurath MF.Apoptosis of T cells and the control of inflammatory bowel disease:therapeutic implications[J].Gut,2007,56(2):293-303.
[4]Zhou PQ,Fan H,Hu H,et al.Role of DOR-β-arrestin1-Bcl2signal transduction pathway and intervention effects of oxymatrine in ulcerative colitis[J].J Huazhong Univ Sci Technolog Med Sci,2014,34(6):815-820.
[5]Fan H,Liu XX,Zhang LJ,et al.Intervention effects of QRZSLXF,a Chinese medicinal herb recipe,on the DOR-β-arrestin1-Bcl2 signal transduction pathway in a rat model of ulcerative colitis[J].Ethnopharmacol,2014,154(1):88-97.
[6]Shi Y,Feng Y,Kang J,et al.Critical regulation of CD4+T cell survival and autoimmunity by beta-arrestin 1[J].Nat Immunol,2007,8(8):817-824.
[7]Kucharzik T,Maaser C,Lügering A,et al.Recent understanding of IBD pathogenesis:implications for future therapies[J].Inflamm Bowel Dis,2006,12(11):1068-1083.
[8]王飞,王建民,石健.肠炎Ⅰ号灌肠方保留灌肠治疗溃疡性直肠炎30例[J].中医研究,2012,25(1):16-18.
[9]李明,王建民,石健,等.肠炎清Ⅰ号合锡类散保留灌肠治疗湿热内蕴型溃疡性结肠炎15例[J].安徽中医学院学报,2012,31(3):34-37.
[10]张冰冰,齐越,贾冬,等.2,4,6-三硝基苯磺酸诱导溃疡性结肠炎大鼠模型的建立及评价[J].中华中医药学刊,2015,33(8):1834-1836.
[11]贺海辉,沈洪,朱宣宣,等.2,4,6-三硝基苯磺酸/乙醇法诱导建立溃疡性结肠炎大鼠模型[J].中国老年学杂志,2015,35(15):4138-4140.
[12]李斌,谷松.乌梅丸及其拆方对溃疡性结肠炎大鼠结肠黏膜组织IL-10与TNF-α的影响[J].辽宁中医药大学学报,2015,17(6):20-22.
[13]王玉芳,魏兵,欧阳钦.转化生长因子β1在溃疡性结肠炎中的表达[J].四川大学学报(医学版),2005,36(2):204-206.
[14]Bickerstaff A,Orosz C.Evidence for a limited contribution of immune regulation to cardiac allograft acceptance[J].Hum Immunol,2002,63(10):935-947.
[15]刘健,杨小玉,董健,等.Bcl-2蛋白与细胞凋亡的关系及其在中枢神经损伤修复中作用的研究进展[J].吉林大学学报(医学版),2012,38(3):603-606.
[16]Kang J,Shi Y,Xiang B,et al.A nuclear function of beta-arrestin1 in GPCR signaling:regulation of histone acetylation and gene transcription[J].Cel,2005,123(5):833-847.
[17]Martinou JC,Youle RJ.Mitochondria in apoptosis:Bcl-2 family members and mitochondrial dynamics[J].Dev Cell,2011,21(1):92-101.
[18]孙仁山,陈晓红,程天民,等.白芨对大鼠创面愈合几个要素的影响[J].中国临床康复,2003,7(29):3927-3929.
[19]何英,李春明,赵巍,等.锡类散治疗溃疡性结肠炎疗效观察[J].中国临床医学,2011,18(3):347-348.