黄连素激活Nrf2通路缓解新生大鼠哮喘模型氧化应激反应和对心肺组织的保护作用
|
摘要
目的:探讨黄连素(berberine,BB)对新生大鼠哮喘模型氧化应激反应的影响和对心肺组织的保护作用以及潜在的分子机制。方法:将新生SD雄性大鼠随机分成健康对照组、健康加药组、哮喘模型组和模型加药组;卵清蛋白诱导新生大鼠哮喘模型;收集肺泡灌洗液(broncho alveokar lavage fluid,BALF)并分析细胞类型和细胞数目;苏木伊红(hematoxylin eosin,HE)染色观察心肌组织和肺组织病理变化;末端标记法(terminal deoxynucleoitidyl transferase mediated nick end labeling,TUNEL)染色检测心肌组织和肺组织细胞凋亡情况;试剂盒检测血清中超氧化物歧化酶(superoxide dismutase,SOD)的活性和丙二醛(malondialdehyde,MDA)、一氧化氮(nitric oxide,NO)的含量;蛋白质印记检测肺组织中Kelch样ECH相关蛋白1(Kelch-like ECH-associated protein 1,Keap1)、NFE2相关因子2(NF-E2 related factor 2,Nrf2)和血红素氧合酶1(heme oxygenase 1,HMOX-1)的表达水平。结果:哮喘模型组与对照组相比,肺泡灌洗液中炎性细胞的数目显著增加;心肌组织细胞排列不规则,部分细胞结构不清晰;肺组织中有大量的炎性细胞浸润,黏膜下水肿,气道上皮断裂脱落,支气管壁明显增厚;心肌组织和肺组织中凋亡细胞比例显著增加;血清中SOD的活性显著降低,MDA和NO的含量显著增加;肺组织中Keap1、Nrf2和HMOX-1的表达水平显著升高。模型加药组与模型组相比,肺泡灌洗液中炎性细胞的数目明显减少;心肌组织较规则,细胞排列较整齐;肺组织中仍有部分炎性细胞浸润,部分肺泡壁增厚;心肌组织和肺组织中凋亡细胞比例显著降低;SOD的活性显著升高,MDA和NO的含量显著降低;Keap1、Nrf2和HMOX-1的表达水平显著升高。结论:黄连素可激活Nrf2通路,缓解新生大鼠哮喘模型的氧化应激反应,对心肺组织具有保护作用。
OBJECTIVE To investigate the potential cardiopulmonary protective effects of berberine on oxidative stress in the asthmatic neonatal rat model,and to explore the potential molecular mechanism.METHODS Male neonatal SD rats were randomly divided into the healthy control group,healthy medication group,asthma model group and model medication group. Ovalbumin was used to induce the newborn asthma rat model; Broncho alveolar lavage fluid(BALF) was collected,and cell type and number were analyzed. Pathological changes in the myocardial tissues and lung tissues were observed through hematoxylin eosin staining. Apoptosis of cardiomyocytes and lung tissues was detected with TUNEL staining; superoxide dismutase(SOD) activity and contents of malondialdehyde(MDA) and nitric oxide(NO) in the serum were detected with related kits. The expression levels of Kelch-like ECH-associated protein 1(Keap1),NFE2 related factor 2(Nrf2) and heme oxygenase 1(HMOX-1) in lung tissues were detected with Western blot. RESULTS Compared with the control group,the number of inflammatory cells in the alveolar lavage fluid increased significantly in the asthma model group. The myocardial tissue cells showed irregular arrangement and some cell structures were not clear. Infiltration of a large number of inflammatory cells was observed in lung tissues,with submucosal edema,airway epithelium rupture and shedding,and significantly thickened bronchial wall. The ratio of apoptotic cells in lung tissues increased significantly. The activity of SOD in the serum decreased significantly,and the content of MDA and NO increased significantly. And the expression levels of Keap1,Nrf2 and HMOX-1 in lung tissues increased significantly. Compared with the model group,the number of inflammatory cells in the alveolar lavage fluid in the model group was significantly reduced; the myocardium was more regular and the cells were arranged more regularly; some inflammatory cell infiltration in the lung tissue and some alveolar wall thickening still existed. The apoptotic cell ratio in the myocardium and lung tissues was significantly decreased. The activity of SOD was significantly increased,and the concentrations of MDA and NO were significantly reduced. The expression levels of Keap1,Nrf2 and HMOX-1 increased significantly.CONCLUSION Berberine can activate the Nrf2 pathway and alleviate oxidative stress in the neonatal asthma rat model,so as to protect cardiopulmonary tissues.
OBJECTIVE To investigate the potential cardiopulmonary protective effects of berberine on oxidative stress in the asthmatic neonatal rat model,and to explore the potential molecular mechanism.METHODS Male neonatal SD rats were randomly divided into the healthy control group,healthy medication group,asthma model group and model medication group. Ovalbumin was used to induce the newborn asthma rat model; Broncho alveolar lavage fluid(BALF) was collected,and cell type and number were analyzed. Pathological changes in the myocardial tissues and lung tissues were observed through hematoxylin eosin staining. Apoptosis of cardiomyocytes and lung tissues was detected with TUNEL staining; superoxide dismutase(SOD) activity and contents of malondialdehyde(MDA) and nitric oxide(NO) in the serum were detected with related kits. The expression levels of Kelch-like ECH-associated protein 1(Keap1),NFE2 related factor 2(Nrf2) and heme oxygenase 1(HMOX-1) in lung tissues were detected with Western blot. RESULTS Compared with the control group,the number of inflammatory cells in the alveolar lavage fluid increased significantly in the asthma model group. The myocardial tissue cells showed irregular arrangement and some cell structures were not clear. Infiltration of a large number of inflammatory cells was observed in lung tissues,with submucosal edema,airway epithelium rupture and shedding,and significantly thickened bronchial wall. The ratio of apoptotic cells in lung tissues increased significantly. The activity of SOD in the serum decreased significantly,and the content of MDA and NO increased significantly. And the expression levels of Keap1,Nrf2 and HMOX-1 in lung tissues increased significantly. Compared with the model group,the number of inflammatory cells in the alveolar lavage fluid in the model group was significantly reduced; the myocardium was more regular and the cells were arranged more regularly; some inflammatory cell infiltration in the lung tissue and some alveolar wall thickening still existed. The apoptotic cell ratio in the myocardium and lung tissues was significantly decreased. The activity of SOD was significantly increased,and the concentrations of MDA and NO were significantly reduced. The expression levels of Keap1,Nrf2 and HMOX-1 increased significantly.CONCLUSION Berberine can activate the Nrf2 pathway and alleviate oxidative stress in the neonatal asthma rat model,so as to protect cardiopulmonary tissues.
引文
[1] Katikireddi SV.Global,regional,and national incidence and prevalence,and years lived with disability for 328 diseases and injuries in 195 countries,1990-2016:A systematic analysis for the global burden of disease study 2016 [J].Lancet,2017,390(10100):1211-1259.
[2] Reid J,Marciniuk DD,Cockcroft DW.Asthma management in the emergency department [J].Can Respir J,2016,7(3):255-260.
[3] Bayes HK,Cowan DC.Biomarkers and asthma management:An update [J].Curr Opin Allergy Clin Immunol,2016,16(3):210-217.
[4] Wen SQ,Jeyakkumar P,Avula SR,et al.Discovery of novel berberine imidazoles as safe antimicrobial agents by down regulating ros generation [J].Bioorg Med Chem Lett,2016,26(12):2768-2773.
[5] Zhang Z,Li X,Li F,et al.Berberine alleviates postoperative cognitive dysfunction by suppressing neuroinflammation in aged mice [J].Int Immunopharmacol,2016,38:426-433.
[6] Chen DL,Yang KY.Berberine alleviates oxidative stress in islets of diabetic mice by inhibiting mir-106b expression and up-regulating sirt1 [J].J Cell Biochem,2017,118(12):4349-4357.
[7] Yao BY,Huang ZM.Berberine protect mice againest CCl4-induced acute liver injury[J].Chin J Mod Appl Pharm(中国现代应用药学),2016,33(4):424-427.
[8] Li Z,Zheng J,Zhang N.Berberine improves airway inflammation and inhibits nf-κb signaling pathway in an ovalbumin-induced rat model of asthma [J].J Asthma,2016,53(10):999-1005.
[9] Kim SJ,Kim CH,Ahn JH,et al.Time sequence of airway remodeling in a mouse model of chronic asthma:The relation with airway hyperresponsiveness [J].J Korean Med Sci,2007,22(2):183-191.
[10] Hou CC,Zhao HJ,Cai SX,et al.Expression of high mobility group box-1 in the lung tissue and balf of asthmatic mice and the influence of dexamethasone[J].J South Med Univ(南方医科大学学报),2010,30(9):2051-2054.
[11] Lin K,Liu S,Shen Y,et al.Berberine attenuates cigarette smoke-induced acute lung inflammation [J].Inflammation,2013,36(5):1079-1086.
[12] Zhang X,Liang D,Lian X,et al.Berberine activates nrf2 nuclear translocation and inhibits apoptosis induced by high glucose in renal tubular epithelial cells through a phosphatidylinositol 3-kinase/akt-dependent mechanism [J].Apoptosis,2016,21(6):721-736.
[13] Lin X,Zhang N.Berberine:Pathways to protect neurons [J].Phytotherapy Res,2018,32(8):1501-1510.
[14] Zhu X,Guo X,Mao G,et al.Hepatoprotection of berberine against hydrogen peroxide-induced apoptosis by upregulation of sirtuin 1 [J].Phytotherapy Res,2013,27(3):417-421.
[15] Kleniewska P,Pawliczak R.The participation of oxidative stress in the pathogenesis of bronchial asthma [J].Biomed Pharmacother,2017,94:100-108.
[16] Mcgovern T,Goldberger M,Chen M,et al.Cyslt1 receptor is protective against oxidative stress in a model of irritant-induced asthma [J].J Immunol,2016,197(1):266-277.
[17] Janssen-Heininger Y,Ckless K,Reynaert N,et al.Sod inactivation in asthma:Bad news or no news?[J].Am J Pathol,2005,166(3):649-652.
[18] Sugiura H,Ichinose M.Role of oxidative stress in aggravation of asthma [J].Arerugi,2017,66(7):931-935.
[19] Kaspar JW,Niture SKJaiswal AK.Nrf2:Inrf2 (keap1) signaling in oxidative stress [J].Free Radic Biol Med,2009,47(9):1304-1309.
[20] Kensler TW,Wakabayashi N,Biswal S.Cell survival responses to environmental stresses via the keap1-nrf2-are pathway [J].Annu Rev Pharmacol Toxicol,2007,47(1):89-116.
[21] Duan L,Li J,Ma P,et al.Vitamin e antagonizes ozone-induced asthma exacerbation in balb/c mice through the nrf2 pathway [J].Food Chem Toxicol,2017,107(Pt A):47-56.
[22] Nadeem A,Siddiqui N,Al-Harbi NO,et al.Tlr-7 agonist attenuates airway reactivity and inflammation through nrf2-mediated antioxidant protection in a murine model of allergic asthma [J].Int J Biochem Cell Biol,2016,73:53-62.
[23] Wang L,Zhao YL,Liu NN,et al.Epithelial ho-1/stat3 affords the protection of subanesthetic isoflurane against zymosan-induced lung injury in mice [J].Oncotarget,2017,8(33):54889-54903.
[24] Leung WS,Yang ML,Lee SS,et al.Protective effect of zerumbone reduces lipopolysaccharide-induced acute lung injury via antioxidative enzymes and nrf2/ho-1 pathway [J].Int Immunopharmacol,2017,46:194-200.
[2] Reid J,Marciniuk DD,Cockcroft DW.Asthma management in the emergency department [J].Can Respir J,2016,7(3):255-260.
[3] Bayes HK,Cowan DC.Biomarkers and asthma management:An update [J].Curr Opin Allergy Clin Immunol,2016,16(3):210-217.
[4] Wen SQ,Jeyakkumar P,Avula SR,et al.Discovery of novel berberine imidazoles as safe antimicrobial agents by down regulating ros generation [J].Bioorg Med Chem Lett,2016,26(12):2768-2773.
[5] Zhang Z,Li X,Li F,et al.Berberine alleviates postoperative cognitive dysfunction by suppressing neuroinflammation in aged mice [J].Int Immunopharmacol,2016,38:426-433.
[6] Chen DL,Yang KY.Berberine alleviates oxidative stress in islets of diabetic mice by inhibiting mir-106b expression and up-regulating sirt1 [J].J Cell Biochem,2017,118(12):4349-4357.
[7] Yao BY,Huang ZM.Berberine protect mice againest CCl4-induced acute liver injury[J].Chin J Mod Appl Pharm(中国现代应用药学),2016,33(4):424-427.
[8] Li Z,Zheng J,Zhang N.Berberine improves airway inflammation and inhibits nf-κb signaling pathway in an ovalbumin-induced rat model of asthma [J].J Asthma,2016,53(10):999-1005.
[9] Kim SJ,Kim CH,Ahn JH,et al.Time sequence of airway remodeling in a mouse model of chronic asthma:The relation with airway hyperresponsiveness [J].J Korean Med Sci,2007,22(2):183-191.
[10] Hou CC,Zhao HJ,Cai SX,et al.Expression of high mobility group box-1 in the lung tissue and balf of asthmatic mice and the influence of dexamethasone[J].J South Med Univ(南方医科大学学报),2010,30(9):2051-2054.
[11] Lin K,Liu S,Shen Y,et al.Berberine attenuates cigarette smoke-induced acute lung inflammation [J].Inflammation,2013,36(5):1079-1086.
[12] Zhang X,Liang D,Lian X,et al.Berberine activates nrf2 nuclear translocation and inhibits apoptosis induced by high glucose in renal tubular epithelial cells through a phosphatidylinositol 3-kinase/akt-dependent mechanism [J].Apoptosis,2016,21(6):721-736.
[13] Lin X,Zhang N.Berberine:Pathways to protect neurons [J].Phytotherapy Res,2018,32(8):1501-1510.
[14] Zhu X,Guo X,Mao G,et al.Hepatoprotection of berberine against hydrogen peroxide-induced apoptosis by upregulation of sirtuin 1 [J].Phytotherapy Res,2013,27(3):417-421.
[15] Kleniewska P,Pawliczak R.The participation of oxidative stress in the pathogenesis of bronchial asthma [J].Biomed Pharmacother,2017,94:100-108.
[16] Mcgovern T,Goldberger M,Chen M,et al.Cyslt1 receptor is protective against oxidative stress in a model of irritant-induced asthma [J].J Immunol,2016,197(1):266-277.
[17] Janssen-Heininger Y,Ckless K,Reynaert N,et al.Sod inactivation in asthma:Bad news or no news?[J].Am J Pathol,2005,166(3):649-652.
[18] Sugiura H,Ichinose M.Role of oxidative stress in aggravation of asthma [J].Arerugi,2017,66(7):931-935.
[19] Kaspar JW,Niture SKJaiswal AK.Nrf2:Inrf2 (keap1) signaling in oxidative stress [J].Free Radic Biol Med,2009,47(9):1304-1309.
[20] Kensler TW,Wakabayashi N,Biswal S.Cell survival responses to environmental stresses via the keap1-nrf2-are pathway [J].Annu Rev Pharmacol Toxicol,2007,47(1):89-116.
[21] Duan L,Li J,Ma P,et al.Vitamin e antagonizes ozone-induced asthma exacerbation in balb/c mice through the nrf2 pathway [J].Food Chem Toxicol,2017,107(Pt A):47-56.
[22] Nadeem A,Siddiqui N,Al-Harbi NO,et al.Tlr-7 agonist attenuates airway reactivity and inflammation through nrf2-mediated antioxidant protection in a murine model of allergic asthma [J].Int J Biochem Cell Biol,2016,73:53-62.
[23] Wang L,Zhao YL,Liu NN,et al.Epithelial ho-1/stat3 affords the protection of subanesthetic isoflurane against zymosan-induced lung injury in mice [J].Oncotarget,2017,8(33):54889-54903.
[24] Leung WS,Yang ML,Lee SS,et al.Protective effect of zerumbone reduces lipopolysaccharide-induced acute lung injury via antioxidative enzymes and nrf2/ho-1 pathway [J].Int Immunopharmacol,2017,46:194-200.