黄芪多糖抑制p38MAPK信号通路减轻缺氧复氧肾小管上皮细胞炎症反应
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摘要
目的研究黄芪多糖对缺氧复氧肾小管上皮细胞炎症反应的影响及机制。方法构建缺氧复氧肾小管上皮细胞损伤模型,用黄芪多糖处理后,分光光度计法检测上清中乳酸脱氢酶(LDH)含量,ELISA法检测上清中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平,Western blot法检测细胞中磷酸化p38MAPK蛋白水平。用p38MAPK激活剂和黄芪多糖处理缺氧复氧肾小管上皮细胞,同样检测细胞分泌的LDH、IL-1β、TNF-α水平。结果缺氧复氧诱导肾小管上皮细胞培养液上清中LDH水平和IL-1β、TNF-α水平升高,促进细胞中磷酸化p38MAPK蛋白表达(P<0.05)。黄芪多糖处理能够下调缺氧复氧肾小管上皮细胞培养液上清中LDH和IL-1β、TNF-α水平,降低细胞中磷酸化p38MAPK蛋白水平(P<0.05)。p38MAPK激活剂处理可以逆转黄芪多糖对缺氧复氧肾小管上皮细胞分泌LDH、IL-1β、TNF-α的促进作用。结论黄芪多糖能够通过抑制p38MAPK信号通路减轻缺氧复氧肾小管上皮细胞炎症反应。
To study the effects and mechanisms of Astragalus polysaccharides in inflammatory response of renal tubular epithelial cells after hypoxia reoxygenation, a hypoxia reoxygenation injury model of renal tubular epithelial cells was constructed and treated with Astragalus polysaccharides. The content of LDH in supernatant was detected by spectrophotometer, while ELISA method was used to detect the levels of IL-1β and TNF-α in the supernatant. Western blot assay was used to detect the level of phosphorylated p38 MAPK protein in cells.Furthermore, both p38 MAPK activator and Astragalus polysaccharides were used to treat anoxia reoxygenation renal tubular epithelial cells, and the levels of LDH, IL-1β and TNF-α secreted by cells were also detected. Data showed that the levels of LDH, IL-1β, TNF-α, and phosphorylated p38 MAPK protein were increased in the supernatant of hypoxia reoxygenation renal tubular epithelial cells(P<0.05), and Astragalus polysaccharide treatment could downregulate the indexes mentioned above(P<0.05). p38 MAPK activator could reverse the effects of Astragalus polysaccharide on LDH, IL-1β and TNF-α secretion in hypoxic-reoxygenated renal tubular epithelial cells.In conclusion, Astragalus polysaccharide can alleviate the inflammation of renal tubular epithelial cells by inhibiting p38 MAPK signaling pathway.
To study the effects and mechanisms of Astragalus polysaccharides in inflammatory response of renal tubular epithelial cells after hypoxia reoxygenation, a hypoxia reoxygenation injury model of renal tubular epithelial cells was constructed and treated with Astragalus polysaccharides. The content of LDH in supernatant was detected by spectrophotometer, while ELISA method was used to detect the levels of IL-1β and TNF-α in the supernatant. Western blot assay was used to detect the level of phosphorylated p38 MAPK protein in cells.Furthermore, both p38 MAPK activator and Astragalus polysaccharides were used to treat anoxia reoxygenation renal tubular epithelial cells, and the levels of LDH, IL-1β and TNF-α secreted by cells were also detected. Data showed that the levels of LDH, IL-1β, TNF-α, and phosphorylated p38 MAPK protein were increased in the supernatant of hypoxia reoxygenation renal tubular epithelial cells(P<0.05), and Astragalus polysaccharide treatment could downregulate the indexes mentioned above(P<0.05). p38 MAPK activator could reverse the effects of Astragalus polysaccharide on LDH, IL-1β and TNF-α secretion in hypoxic-reoxygenated renal tubular epithelial cells.In conclusion, Astragalus polysaccharide can alleviate the inflammation of renal tubular epithelial cells by inhibiting p38 MAPK signaling pathway.
引文
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[20]罗素,荣晓凤,彭菲菲.大黄素对TNF-α诱导的成纤维样滑膜细胞株MH7A细胞ERK1/2和p38MAPK的影响[J].免疫学杂志,2017,33(2):113-117.
[21]何丹,任朋亮,范雪娇,等.p38MAPK与疾病关系的研究进展[J].西部医学,2014,26(2):259-262.
[22]Liu LM,Tu WJ,Zhu T,et al.IRF3 is an important molecule in the UII/UT system and mediates immune inflammatory injury in acute liver failure[J].Oncotarget,2016,7(31):49027-49041.
[23]Schett G,Zwerina J,Firestein G.The p38 mitogenactivated protein kinase(MAPK)pathway in rheumatoid arthritis[J].Ann Rheum Dis,2008,67(7):909-916.
[24]Li Z,Wang H,Liu L.Interleukin-25 enhances allergic inflammation through p38MAPK and NF-κB pathways in mouse models of allergic rhinitis[J].Iran J Allergy Asthma Immunol,2014,13(6):412-419.
[25]刘蓓,朱海燕,高永红,等.黄芪多糖对TNF-α诱导心脏微血管内皮细胞黏附分子基因转录及p38MAPK信号通路的影响[J].世界中医药,2011,6(3):263-265.
[2]Ma T,Huang C,Xu Q,et al.Suppression of BMP-7 by histone deacetylase 2 promoted apoptosis of renal tubular epithelial cells in acute kidney injury[J].Cell Death Dis,2017,8(10):e3139.
[3]Kasprowska D,Machnik G,Kost A,et al.Time-dependent changes in apoptosis upon autophagy inhibition in astrocytes exposed to oxygen and glucose deprivation[J].Cell Mol Neurobiol,2017,37(2):1-12.
[4]黄家林,张勇.黄芪多糖抗炎免疫作用机制研究进展[J].中西医结合心脑血管病杂志,2013,11(11):1374-1376.
[5]宋艳,丁国华,桂元,等.黄芪多糖对肾盂肾炎大鼠肾脏TLR4表达的影响[J].武汉大学学报(医学版),2006,27(5):605-608.
[6]Zhang YW,Wu CY,Cheng JT.Merit of Astragalus polysaccharide in the improvement of early diabetic nephropathy with an effect on mRNA expressions of NF-kappaB and IkappaB in renal cortex of streptozotoxininduced diabetic rats[J].J Ethnopharmacol,2007,114(3):387-392.
[7]李智军,魏连波,贺丰,等.黄芪多糖治疗大鼠系膜细胞增生性肾炎的实验研究[J].中国中西医结合肾病杂志,2000,1(4):206-208.
[8]杨桂菊,张东霞.黄芪多糖通过p38MAPK-HSP27通路减轻大鼠急性坏死性胰腺炎肺损伤[J].中国中医急症,2017,26(12):2093-2096.
[9]赵启明,李彩娥,雷蕾,等.黄芪多糖在未成年大鼠心肌缺血再灌注炎性损伤中的作用[J].中国医院药学杂志,2017,37(7):625-628.
[10]Zhang L,Wang Y,Ma J,et al.Exogenous MSCs ameliorate hypoxia/reoxygenation injury in renal tubular epithelial cells through JAK/STAT signaling pathway-mediated regulation of HMGB1[J].Am J Transl Res,2017,9(5):2412-2420.
[11]Fontana J,Vogt A,Hohenstein A,et al.Impact of steroids on the inflammatory response after ischemic acute kidney injury in rats[J].Indian J Nephrol,2017,27(5):365-371.
[12]陈英,张文玲,黄涛,等.炎症因子TNF-α、IL-6、IL-17与类风湿关节炎并发动脉粥样硬化的关系[J].免疫学杂志,2017,33(3):268-272.
[13]Wang G,Cui J,Guo Y,et al.Cyclosporin a protects H9c2cells against chemical hypoxia-induced injury via inhibition of MAPK signaling pathway[J].Int Heart J,2016,57(4):483-489.
[14]李红专.黄芪对糖尿病肾病的药理作用机制研究进展[J].中国社区医师(医学专业),2012,14(15):6-7.
[15]李飞飞,沈建平,庄海峰.黄芪多糖对免疫作用的研究进展[J].医学综述,2012,18(8):1227-1229.
[16]Zhou X,Xin Q,Wang Y,et al.Total flavonoids of astragalus plays a cardioprotective role in viral myocarditis[J].Acta Cardiol Sin,2016,32(1):81-88.
[17]Cui K,Zhang S,Jiang X,et al.Novel synergic antidiabetic effects of Astragalus polysaccharides combined with Crataegus flavonoids via improvement of islet function and liver metabolism[J].Mol Med Rep,2016,13(6):4737-4744.
[18]刘爽,刘晓玲,张勇,等.黄芪多糖对CVB3感染的心肌细胞和心肌的保护作用研究[J].中国分子心脏病学杂志,2015,15(4):1407-1411.
[19]房信胜,穆象山,周红英,等.黄芪皂苷和黄芪多糖对大鼠肾脏系膜细胞增殖的影响[J].时珍国医国药,2008,19(6):1448-1449.
[20]罗素,荣晓凤,彭菲菲.大黄素对TNF-α诱导的成纤维样滑膜细胞株MH7A细胞ERK1/2和p38MAPK的影响[J].免疫学杂志,2017,33(2):113-117.
[21]何丹,任朋亮,范雪娇,等.p38MAPK与疾病关系的研究进展[J].西部医学,2014,26(2):259-262.
[22]Liu LM,Tu WJ,Zhu T,et al.IRF3 is an important molecule in the UII/UT system and mediates immune inflammatory injury in acute liver failure[J].Oncotarget,2016,7(31):49027-49041.
[23]Schett G,Zwerina J,Firestein G.The p38 mitogenactivated protein kinase(MAPK)pathway in rheumatoid arthritis[J].Ann Rheum Dis,2008,67(7):909-916.
[24]Li Z,Wang H,Liu L.Interleukin-25 enhances allergic inflammation through p38MAPK and NF-κB pathways in mouse models of allergic rhinitis[J].Iran J Allergy Asthma Immunol,2014,13(6):412-419.
[25]刘蓓,朱海燕,高永红,等.黄芪多糖对TNF-α诱导心脏微血管内皮细胞黏附分子基因转录及p38MAPK信号通路的影响[J].世界中医药,2011,6(3):263-265.