NADPH氧化酶抑制剂Apocynin预防脂多糖心肌纤维化的研究
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摘要
目的探讨NADPH氧化酶抑制剂Apocynin对脂多糖(Lipopolysaccharide,LPS)心肌纤维化的预防作用。方法 40只小鼠随机分成4组:对照组(CON)、Apocynin组、LPS组和LPS+Apocynin组,每组10只。LPS组和LPS+Apocynin组给予LPS 10 mg/kg,每周1次,连续4周;Apocynin组和LPS+Apocynin组给予Apocynin 10 mg/kg,1次/d,连续4周。采用天狼星红染色评估心肌纤维化程度,RT-PCR检测ColⅠa1、ColⅢa1、MMP2、MMP9 m RNA的表达,Western blot检测心肌gp91~(phox) 和p67~(phox)的表达,DHE检测心肌活性氧生成。结果小鼠LPS注射4周后出现心肌纤维化。与对照组比较,LPS组心肌间质内弥漫性纤维组织显著增多,胶原容积分数增加,心肌组织中胶原相关因子ColⅠa1、ColⅢa1、MMP2、MMP9 m RNA表达明显增加,心肌gp91~(phox) 和p67~(phox)的表达增加,ROS生成增加(P<0.05)。与LPS组比较,Apocynin干预可抑制上述指标的增加(P<0.05)。结论 Apocynin能预防LPS诱导小鼠心肌纤维化,其效应可能与抑制心肌gp91~(phox) 和p67~(phox)过表达有关。
Objective To investigate the preventive effects of apocynin,an NADPH oxidase inhibitor,on cardiac fibrosis in mice exposed to subclinical lipopolysaccharide. Methods Totally 40 male C57 BL/6 J mice w ere divided randomly into 4 groups: control group,apocynin group,LPS group and LPS + apocynin group. LPS w as injected intraperitoneally once a w eek for 4 w eeks to mimic cardiac fibrosis in LPS group and LPS + apocynin group. Apocynin w as given in apocynin group and LPS + apocynin group,once a w eek for 4 w eeks. Collagen fraction area of the left ventricle w as measured w ith picrosirius red staining. The cardiac expression of fibrosis-related genes w as measured by RT-PCR. The subunits gp91~(phox) and p67~(phox)of NADPH oxidase w ere analyzed by Western blot,ROS production w as determined by DHE staining. Results The cardiac fibrosis of mice occurred after 4 w eeks. Compared w ith control group,the interstitial diffuse fibrous tissue in myocardium,the collagen volume fraction,the m RNA expression of LPS-induced activation of fibrosis-related genes ColⅠa1,ColⅢa1,M M P2 and M M P9,the expression gp91~(phox) and p67~(phox)and ROS production in LPS group w ere increased. Compared w ith LPS group,apocynin decreased the above indexes significantly( P < 0. 05). Conclusion Apocynin can protect against the cardiac fibrosis in mice exposed to subclinical LPS by inhibiting the over-expression of myocardial gp91~(phox) and p67~(phox).
Objective To investigate the preventive effects of apocynin,an NADPH oxidase inhibitor,on cardiac fibrosis in mice exposed to subclinical lipopolysaccharide. Methods Totally 40 male C57 BL/6 J mice w ere divided randomly into 4 groups: control group,apocynin group,LPS group and LPS + apocynin group. LPS w as injected intraperitoneally once a w eek for 4 w eeks to mimic cardiac fibrosis in LPS group and LPS + apocynin group. Apocynin w as given in apocynin group and LPS + apocynin group,once a w eek for 4 w eeks. Collagen fraction area of the left ventricle w as measured w ith picrosirius red staining. The cardiac expression of fibrosis-related genes w as measured by RT-PCR. The subunits gp91~(phox) and p67~(phox)of NADPH oxidase w ere analyzed by Western blot,ROS production w as determined by DHE staining. Results The cardiac fibrosis of mice occurred after 4 w eeks. Compared w ith control group,the interstitial diffuse fibrous tissue in myocardium,the collagen volume fraction,the m RNA expression of LPS-induced activation of fibrosis-related genes ColⅠa1,ColⅢa1,M M P2 and M M P9,the expression gp91~(phox) and p67~(phox)and ROS production in LPS group w ere increased. Compared w ith LPS group,apocynin decreased the above indexes significantly( P < 0. 05). Conclusion Apocynin can protect against the cardiac fibrosis in mice exposed to subclinical LPS by inhibiting the over-expression of myocardial gp91~(phox) and p67~(phox).
引文
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[7]Kim SY,Lee JG,Cho WS,et al.Role of NADPH oxidase-2 in lipopolysaccharide-induced matrix metalloproteinase expression and cell migration[J].Immunol Cell Biol,2010,88(2):197-204.
[8]Ngkelo A,Meja K,Yeadon M,et al.LPS induced inflammatory responses in human peripheral blood mononuclear cells is mediated through NOX4 and Gia dependent PI-3 kinase signalling[J].J Inflamm(Lond),2012,9(1):1.
[9]Gandhirajan RK,Meng S,Chandramoorthy HC,et al.Blockade of NOX2 and STIM1 signaling limits lipopolysaccharide-induced vascular inflammation[J].J Clin Invest,2013,123(2):887-890.
[10]Peng T,Lu X,Feng Q.Pivotal role of gp91phox-containing NADH oxidase in lipopolysaccharide-induced tumor necrosis factor-alpha expression and myocardial depression[J].Circulation,2005,111(13):1637-1644.
[11]Tsutsui H,Kinugawa S,Matsushima S.Oxidative stress and heart failure[J].Am J Physiol Heart Circ Physiol,2011,301(6):H2181-H2190.
[12]Octavia Y,Brunner-La Rocca HP,Moens AL.NADPH oxidase-dependent oxidative stress in the failing heart:from pathogenic roles to therapeutic approach[J].Free Radic Biol M ed,2012,52(2):291-297.
[2]Lew WY,Bayna E,Dalle ME,et al.Myocardial fibrosis induced by exposure to subclinical lipopolysaccharide is associated w ith decreased mi R-29c and enhanced NOX2 expression in mice[J].PLo S One,2014,9(9):e107556.
[3]Johar S,Cave AC,Narayanapanicker A,et al.Aldosterone mediates angiotensin II-induced interstitial cardiac fibrosis via a Nox2-containing NADPH oxidase[J].FASEB J,2006,20(9):1546-1548.
[4]Zhao QD,Viswanadhapalli S,Williams P,et al.NADPH oxidase 4 induces cardiac fibrosis and hypertrophy through activating Akt/m TOR and NF-κB signaling pathw ays[J].Circulation,2015,131(7):643-655.
[5]Abdelmageed ME,El-Awady MS,Suddek GM.Apocynin ameliorates endotoxin-induced acute lung injury in rats[J].Int Immunopharmacol,2016,30:163-170.
[6]Adler V,Yin Z,Tew KD,et al.Role of redox potential and reactive oxygen species in stress signaling[J].Oncogene,1998,18(45):6104-6111.
[7]Kim SY,Lee JG,Cho WS,et al.Role of NADPH oxidase-2 in lipopolysaccharide-induced matrix metalloproteinase expression and cell migration[J].Immunol Cell Biol,2010,88(2):197-204.
[8]Ngkelo A,Meja K,Yeadon M,et al.LPS induced inflammatory responses in human peripheral blood mononuclear cells is mediated through NOX4 and Gia dependent PI-3 kinase signalling[J].J Inflamm(Lond),2012,9(1):1.
[9]Gandhirajan RK,Meng S,Chandramoorthy HC,et al.Blockade of NOX2 and STIM1 signaling limits lipopolysaccharide-induced vascular inflammation[J].J Clin Invest,2013,123(2):887-890.
[10]Peng T,Lu X,Feng Q.Pivotal role of gp91phox-containing NADH oxidase in lipopolysaccharide-induced tumor necrosis factor-alpha expression and myocardial depression[J].Circulation,2005,111(13):1637-1644.
[11]Tsutsui H,Kinugawa S,Matsushima S.Oxidative stress and heart failure[J].Am J Physiol Heart Circ Physiol,2011,301(6):H2181-H2190.
[12]Octavia Y,Brunner-La Rocca HP,Moens AL.NADPH oxidase-dependent oxidative stress in the failing heart:from pathogenic roles to therapeutic approach[J].Free Radic Biol M ed,2012,52(2):291-297.