坎地沙坦对脂多糖刺激巨噬细胞炎症因子释放的影响
|
摘要
目的研究坎地沙坦对脂多糖(LPS)刺激巨噬细胞炎症因子释放的影响。方法 MTT法测定不同浓度坎地沙坦对RAW264.7细胞活性的影响。将细胞分为5组:对照组、LPS干预组、LPS+坎地沙坦10~(-7)mol/L组、LPS+坎地沙坦10~(-6)mol/L组和LPS+坎地沙坦10~(-5)mol/L组。real-time RT-PCR和Western-blot测定各组细胞Toll样受体4(TLR4)、髓样分化因子88(Myd88)和核转录因子NF-κB(p65)mRNA和蛋白的表达;ELISA测定各组细胞上清液IL-1β和TNF-α分泌水平。结果同对照组相比,坎地沙坦可以有效抑制LPS诱导的TLR4、MyD88和NF-κB(p65)的mRNA表达和致炎细胞因子的释放,并具有剂量依赖性(P <0.01)。结论坎地沙坦能够减少巨噬细胞炎症因子IL-1β和TNF-α的分泌,这一作用可能部分通过抑制TLR4-Myd88-NF-κB传导通路的活化得以实现。
Objective To explore the influence of candesartan on the inflammatory cytokine expression of macrophage stimulated by Lipopolysaccharides(LPS). Methods Murine macrophage line(RAW264.7 cell)were stimulated with different concentration candesartan for 24 h,and then cell activity was observed by MTT assay.Cells were pretreated with different concentration candesartan(10~(-7),10~(-6) and 10~(-5)μmol/L)for 1 h,and then stimulated with LPS(500 ng/mL)for 4,9 or 24 h. mRNA and protein of Toll-like receptor-4(TLR4),myeloid differentiation primary response gene 88(Myd88)and Nuclear factor κB(NF-κB)were determined by real-time PCR and Western-blot;IL-1βand TNF-α concentration were detected by ELISA. Results Candesartan effectively inhibited mRNA and protein expressions of TLR4,Myd88 and NF-κB(p65)and secretion of IL-1βand TNF-α in macrophage stimulated by LPS,with concentration-dependent manners. Conclusion Candesartan has inhibitory dependent manner in macrophage stimulated by LPS,and this effect was also partly associated with inactivity of TLR4-Myd88-NF-κB signaling pathway.
Objective To explore the influence of candesartan on the inflammatory cytokine expression of macrophage stimulated by Lipopolysaccharides(LPS). Methods Murine macrophage line(RAW264.7 cell)were stimulated with different concentration candesartan for 24 h,and then cell activity was observed by MTT assay.Cells were pretreated with different concentration candesartan(10~(-7),10~(-6) and 10~(-5)μmol/L)for 1 h,and then stimulated with LPS(500 ng/mL)for 4,9 or 24 h. mRNA and protein of Toll-like receptor-4(TLR4),myeloid differentiation primary response gene 88(Myd88)and Nuclear factor κB(NF-κB)were determined by real-time PCR and Western-blot;IL-1βand TNF-α concentration were detected by ELISA. Results Candesartan effectively inhibited mRNA and protein expressions of TLR4,Myd88 and NF-κB(p65)and secretion of IL-1βand TNF-α in macrophage stimulated by LPS,with concentration-dependent manners. Conclusion Candesartan has inhibitory dependent manner in macrophage stimulated by LPS,and this effect was also partly associated with inactivity of TLR4-Myd88-NF-κB signaling pathway.
引文
[1]WU M Y,LI C J,HOU M F,et al.New insights into the role of inflammation in the pathogenesis of atherosclerosis[J].Int JMol Sci,2017,18(10):2034.
[2]JIAN-JUN L I.Research progress of inflammation and atherosclerosis[J].Adv Cardiovasc Dis,2012,33(1):10-13.
[3]LINTON M R F,BABAEY V R,HUANG J,et al.Macrophage apoptosis and efferocytosis in the pathogenesis of atherosclerosis[J].J Circ,2016,80(11):2259-2268.
[4]BRUCE A,BEUTLER.TLRs and innate immunity[J].BLOOD,2009,113(7):1399-1407.
[5]O′NEILL L A J,GOLENBOCK D,BOWIE A G.The history of Toll-like receptors redefining innate immunity[J].Nat Rev Immunol,2013,13(6):453-460.
[6]TAKEDA K,AKIRA S.Toll-like receptors in innate immunity[J].Int Immuol,2005,17(1):1-14.
[7]SALTIEL A R,OLEFSKY J M.Inflammatory mechanisms linking obesity and metabolic disease[J].J Clin Invest,2017,127(1):1-4.
[8]ABRAHAM S N,MIAO Y.The nature of immune responses to urinary tract infections[J].Nat Rev Immunol,2015,15(10):655-663.
[9]SAVVA A,ROGER T.Targeting toll-like receptors:promising therapeutic strategies for the management of sepsis-associated pathology and infectious diseases[J].Front Immunol,2013,4:387.
[10]HEENEMAN S,SLUIMER J C,DAEMEN M J A P.Angiotensin converting enzyme and vascular remodeling[J].Circ Res,2007,101:441-454.
[11]WOLF J,ROSE-JOHN S,GARBERS C.Interleukin-6 and its receptors:a highly regulated and dynamic system[J].Cytokine,2014,70(1):11-20.
[12]ARUMUGAM S,SREEDHAR R,THANDAYARAYAN R A,et al.Angiotensin receptor blockers:Focus on cardiac and renal injury[J].Trends Cardiovasc Med,2016,26(3):221-228.
[13]BENICKY J,SANCHEZ-LEMUS E,et al.Anti-inflammatory effects of angiotensin receptor blockersin the brain and the periphery[J].Cell Mol Neurobiol,2009,29(6-7):781-792.
[14]NAIR A R,EENEZER P J,SAINI Y,et al.Angiotensin II-induced hypertensive renal inflammation is mediated through HMGB1-TLR4 signaling in rat tubulo-epithelial cells[J].Exp Cell Res,2015,335(2):238-247
[15]KIKUCHI K,TANCHAROEN S,ITO T,et al.Potential of the angiotensin receptor blockers(ARBs)telmisartan,irbesartan,and candesartan for inhibiting the HMGB1/RAGE axis in prevention and acute treatment of stroke[J].Int J Mol Sci,2013,14(9):18899-18924.
[16]于新辉,闫超,孟哲.坎地沙坦抑制内毒素诱导的VSMCs炎症因子释放的作用研究[J].重庆医学,2014,43(36):4860-4863.
[17]KAWASAKI T,KAWAI T.Toll-like receptor signaling pathways[J].Front Immunol,2014,5:461.
[18]JIMENEZ-DALMARONI M J,GERSWHIN M E,ADAMOPOU-LOS I E.The critical role of toll-like receptors-from microbial recognition to autoimmunity:a comprehensive review[J].Autoimmun Rev,2016,15(1):1-8.
[19]EDFLDT K,SWEDENBORG J,HANSSON G K,et al.Expression of Toll-like receptors in human atherosclerotic lesions:a possible pathway for plaque activation[J].Circulation,2002,105(10):1158-1161.
[20]LI H,SUN B.Toll-like receptor 4 in atherosclerosis[J].J Cell Mol Med,2007,11(1):88-95.
[21]ZHOU Z,DING M,HUANG L,et al.Toll-like receptor-mediated immune responses in intestinal macrophages;implications for mucosal immunity and autoimmune diseases[J].Clin Immunol,2016,173:81-86.
[22]MULLICK A E,TOBIAS P S,CURTISS L K.Toll-like receptors andatherosclerosis:key contributors in disease and health[J].Immunol Res,2006,34(3):193-209.
[23]KAWAI T,AKIRA S.TLR signaling[J].Cell Death Differ,2006,13(5):8156-8165.
[24]MICHEL M C,BRUNNER H R,FOSTER C,et al.Angiotensin II type 1 receptor antagonists in animal models of vascular,cardiac,metabolic and renal disease[J].Pharmacol Ther,2016,164:1-81.
[25]YAN W H,PAN C Y,DOU J T,et al.Candesartan cilexetil prevents diet-induced insulin resistance via peroxisome proliferator-activated receptor-γactivation in an obese rat model[J].Exp Therapeutic Med,2016,12(1):272-278.
[26]DOHI Y,OHASHI M,SUGIYAMA M,et al.Candesartan reduces oxida-tive stress and inflammation in patients with essential hypertension[J].Hypertens Res,2003,26(9):691-697.
[27]LAUKOVA M,VARGOVIC P,ROKYTOVA I,et al.Repeated stress exaggerates lipopolysaccharide-induced inflammatory response in the rat spleen[J].Cell Mol Neurobiol,2018,38(1):195-208.
[2]JIAN-JUN L I.Research progress of inflammation and atherosclerosis[J].Adv Cardiovasc Dis,2012,33(1):10-13.
[3]LINTON M R F,BABAEY V R,HUANG J,et al.Macrophage apoptosis and efferocytosis in the pathogenesis of atherosclerosis[J].J Circ,2016,80(11):2259-2268.
[4]BRUCE A,BEUTLER.TLRs and innate immunity[J].BLOOD,2009,113(7):1399-1407.
[5]O′NEILL L A J,GOLENBOCK D,BOWIE A G.The history of Toll-like receptors redefining innate immunity[J].Nat Rev Immunol,2013,13(6):453-460.
[6]TAKEDA K,AKIRA S.Toll-like receptors in innate immunity[J].Int Immuol,2005,17(1):1-14.
[7]SALTIEL A R,OLEFSKY J M.Inflammatory mechanisms linking obesity and metabolic disease[J].J Clin Invest,2017,127(1):1-4.
[8]ABRAHAM S N,MIAO Y.The nature of immune responses to urinary tract infections[J].Nat Rev Immunol,2015,15(10):655-663.
[9]SAVVA A,ROGER T.Targeting toll-like receptors:promising therapeutic strategies for the management of sepsis-associated pathology and infectious diseases[J].Front Immunol,2013,4:387.
[10]HEENEMAN S,SLUIMER J C,DAEMEN M J A P.Angiotensin converting enzyme and vascular remodeling[J].Circ Res,2007,101:441-454.
[11]WOLF J,ROSE-JOHN S,GARBERS C.Interleukin-6 and its receptors:a highly regulated and dynamic system[J].Cytokine,2014,70(1):11-20.
[12]ARUMUGAM S,SREEDHAR R,THANDAYARAYAN R A,et al.Angiotensin receptor blockers:Focus on cardiac and renal injury[J].Trends Cardiovasc Med,2016,26(3):221-228.
[13]BENICKY J,SANCHEZ-LEMUS E,et al.Anti-inflammatory effects of angiotensin receptor blockersin the brain and the periphery[J].Cell Mol Neurobiol,2009,29(6-7):781-792.
[14]NAIR A R,EENEZER P J,SAINI Y,et al.Angiotensin II-induced hypertensive renal inflammation is mediated through HMGB1-TLR4 signaling in rat tubulo-epithelial cells[J].Exp Cell Res,2015,335(2):238-247
[15]KIKUCHI K,TANCHAROEN S,ITO T,et al.Potential of the angiotensin receptor blockers(ARBs)telmisartan,irbesartan,and candesartan for inhibiting the HMGB1/RAGE axis in prevention and acute treatment of stroke[J].Int J Mol Sci,2013,14(9):18899-18924.
[16]于新辉,闫超,孟哲.坎地沙坦抑制内毒素诱导的VSMCs炎症因子释放的作用研究[J].重庆医学,2014,43(36):4860-4863.
[17]KAWASAKI T,KAWAI T.Toll-like receptor signaling pathways[J].Front Immunol,2014,5:461.
[18]JIMENEZ-DALMARONI M J,GERSWHIN M E,ADAMOPOU-LOS I E.The critical role of toll-like receptors-from microbial recognition to autoimmunity:a comprehensive review[J].Autoimmun Rev,2016,15(1):1-8.
[19]EDFLDT K,SWEDENBORG J,HANSSON G K,et al.Expression of Toll-like receptors in human atherosclerotic lesions:a possible pathway for plaque activation[J].Circulation,2002,105(10):1158-1161.
[20]LI H,SUN B.Toll-like receptor 4 in atherosclerosis[J].J Cell Mol Med,2007,11(1):88-95.
[21]ZHOU Z,DING M,HUANG L,et al.Toll-like receptor-mediated immune responses in intestinal macrophages;implications for mucosal immunity and autoimmune diseases[J].Clin Immunol,2016,173:81-86.
[22]MULLICK A E,TOBIAS P S,CURTISS L K.Toll-like receptors andatherosclerosis:key contributors in disease and health[J].Immunol Res,2006,34(3):193-209.
[23]KAWAI T,AKIRA S.TLR signaling[J].Cell Death Differ,2006,13(5):8156-8165.
[24]MICHEL M C,BRUNNER H R,FOSTER C,et al.Angiotensin II type 1 receptor antagonists in animal models of vascular,cardiac,metabolic and renal disease[J].Pharmacol Ther,2016,164:1-81.
[25]YAN W H,PAN C Y,DOU J T,et al.Candesartan cilexetil prevents diet-induced insulin resistance via peroxisome proliferator-activated receptor-γactivation in an obese rat model[J].Exp Therapeutic Med,2016,12(1):272-278.
[26]DOHI Y,OHASHI M,SUGIYAMA M,et al.Candesartan reduces oxida-tive stress and inflammation in patients with essential hypertension[J].Hypertens Res,2003,26(9):691-697.
[27]LAUKOVA M,VARGOVIC P,ROKYTOVA I,et al.Repeated stress exaggerates lipopolysaccharide-induced inflammatory response in the rat spleen[J].Cell Mol Neurobiol,2018,38(1):195-208.