肉苁蓉苷A通过TGF-β1/VEGF通路对放射性肺炎小鼠肺部氧化应激和炎症指标影响的研究
|
摘要
目的:探究肉苁蓉苷A对放射性肺炎小鼠肺部氧化应激及炎症反应的作用及机制。方法:X射线照射复制小鼠放射性肺损伤模型并同时给予肉苁蓉苷A,30 d后处死小鼠,用Tunel法检测细胞凋亡情况,试剂盒检测肺匀浆抗氧化酶、氧化产物和炎症因子的含量,免疫组化检测TGF-1β的表达,Western blot检测TGF-1β/VEGF通路相关蛋白表达。结果:肉苁蓉苷A能显著抑制放射诱导的凋亡小体的形成,能明显升高肺匀浆抗氧化酶GSH-PX、T-Aoc及SOD的浓度,降低氧化产物MDA浓度;同时,肉苁蓉苷A还能显著降低肺匀浆炎症因子IL-6和IL-1β浓度,促进IL-10分泌。此外,肉苁蓉苷A能显著减少模型小鼠肺组织TGF-β1的阳性表达,降低TGF-β1、VEGF和VEGFR2的蛋白表达水平。结论:肉苁蓉苷A能抑制放射性肺损伤小鼠肺部的氧化应激和炎症反应,其机制与TGF-1β/VEGF通路有关。
Objective: To investigate the effects and mechanism of Cistanoside A( Cis A) on oxidative stress and inflammatory cytokines in radiation-induced pulmonary injury mice. Methods: Mice were exposed to X-ray and were sacrificed after treating with Cis A for 30 days. Cell apoptosis was determined by Tunel assay. The concentrations of antioxidases,an oxidation product and inflammatory cytokines were measured by using kits. Immunohistochemistry was employed to detect the expression of TGF-1β,and the TGF-1β/VEGF pathway related-protein levels were measured by Western blot. Results: Cis A inhibited the formation of apoptotic body significantly,and increased the concentrations of GSH-PX,T-Aoc and SOD coupled with decreasing MDA. Meanwhile,Cis A inhibited inflammation as demonstrated by decreasing IL-6,IL-1β and increasing IL-10. In addition,Cis A inhibited the positive expression of TGF-β1 and down-regulated the protein levels of TGF-β1,VEGF and VEGFR2. Conclusion: Cis A inhibits oxidative stress and inflammation in radiation-induced pulmonary injury mice and the mechanism is related to TGF-β1/VEGF pathway.
Objective: To investigate the effects and mechanism of Cistanoside A( Cis A) on oxidative stress and inflammatory cytokines in radiation-induced pulmonary injury mice. Methods: Mice were exposed to X-ray and were sacrificed after treating with Cis A for 30 days. Cell apoptosis was determined by Tunel assay. The concentrations of antioxidases,an oxidation product and inflammatory cytokines were measured by using kits. Immunohistochemistry was employed to detect the expression of TGF-1β,and the TGF-1β/VEGF pathway related-protein levels were measured by Western blot. Results: Cis A inhibited the formation of apoptotic body significantly,and increased the concentrations of GSH-PX,T-Aoc and SOD coupled with decreasing MDA. Meanwhile,Cis A inhibited inflammation as demonstrated by decreasing IL-6,IL-1β and increasing IL-10. In addition,Cis A inhibited the positive expression of TGF-β1 and down-regulated the protein levels of TGF-β1,VEGF and VEGFR2. Conclusion: Cis A inhibits oxidative stress and inflammation in radiation-induced pulmonary injury mice and the mechanism is related to TGF-β1/VEGF pathway.
引文
[1]Lingos TI,Recht A,Vicini F,et al.Radiation pneumonitis in breast cancer patients treated with conservative surgery and radiation therapy[J].Int J Rad Oncol Biol Phys,1991,21(2):355-360.
[2]Martel MK,Ten Haken RK,Hazuka MB,et al.Dose-volume histogram and 3-D treatment planning evaluation of patients with pneumonitis[J].Int J Rad Oncol Biol Phys,1994,28(3):575-581.
[3]Yu B,Wang J,Xu Y,et al.Radiotherapy techniques and radiation pneumonitis:a lot to a little or a little to a lot?[J].Chin J Lung Cancer,2015,18(12):752-757.
[4]Anscher M S,Garst J,Marks L B,et al.Assessing the ability of the antiangiogenic and anticytokine agent thalidomide to modulate radiation-induced lung injury[J].Int J Rad Oncol Biol Phys,2006,66(2):477-482.
[5]Robbins ME,Diz DI.Pathogenic role of the renin-angiotensin system in modulating radiation-induced late effects[J].Int J Rad Oncol Biol Phys,2006,64(1):6-12.
[6]Lee SJ,Yi C,Heo RW,et al.Clarithromycin attenuates radiationinduced lung injury in mice[J].Plos One,2015,10(6):e0131671.
[7]Antonic V,Rabbani ZN,Jackson IL,et al.Subcutaneous administration of bovine superoxide dismutase protects lungs from radiation-induced lung injury[J].Free Radical Res,2015,49(10):1259-1268.
[8]Abdulla S,Salavati A,Saboury B,et al.Quantitative assessment of global lung inflammation following radiation therapy using FDGPET/CT:a pilot study[J].Eur J Nuclear Med Mol Imaging,2014,41(2):350-356.
[9]Tian W,Chen C,Man Y,et al.Ethanol extract mediates rat sex hormone levels by induction of testicular steroidgenic enzymes[J].Pharmaceutical Biol,2016,54(3):481-487.
[10]Xiong Q,Kadota S,Tani T,et al.Antioxidative effects of phenylethanoids from Cistanche deserticola[J].Biol Pharm Bull,1996,19(12):1580-1585.
[11]Wang T,Zhang X,Xie W.Cistanche deserticola Y.C.Ma,"Desert ginseng":a review[J].Am J Chin Med,2012,40(06):1123-1141.
[12]Gu L,Xiong WT,Wang C,et al.Cistanche deserticola decoction alleviates the testicular toxicity induced by hydroxyurea in male mice[J].Asian J Androl,2013,15(6):838-840.
[13]Luo H,Cao R,Wang L,et al.Protective effect of Cistanchis,A on ethanol-induced damage in primary cultured mouse hepatocytes[J].Biomed Pharmacotherapy,2016,83:1071-1079.
[14]Vujaskovic Z,Marks L B,Anscher M S.The physical parameters and molecular events associated with radiation-induced lung toxicity[J].Seminars Rad Oncol,2000,10(4):296-307.
[15]Nan ZD,Zeng KW,Shi SP,et al.Phenylethanoid glycosides with anti-inflammatory activities from the stems of Cistanche deserticola cultured in Tarim desert[J].Fitoterapia,2013,89(1):167-174.
[16]Haimovitzfriedman A,Yang TJ,Thin TH,et al.Imaging Radiotherapy-Induced Apoptosis[J].Rad Res,2012,177(4):467-482.
[17]Ahire V,Mishra KP,Kulkarni GR.On the mechanism of cellular toxicity in breast cancer by ionizing radiation and chemotherapeutic drugs[J].J Environ Pathol Toxicol Oncol,2014,33(1):69-82.
[18]Bakkal BH,Gultekin FA,Guven B,et al.Effect of ozone oxidative preconditioning in preventing early radiation-induced lung injury in rats[J].Brazilian J Med Biol Res,2013,46(9):789-796.
[19]Martin M,Lefaix J,Delanian S.TGF-beta1 and radiation fibrosis:a master switch and a specific therapeutic target?[J].Int J Rad Oncol Biol Phys,2000,47(2):277-290.
[20]Zaidi A,Jelveh S,Mahmood J,et al.Effects of lipopolysaccharide on the response of C57BL/6J mice to whole thorax irradiation[J].Radiotherapy Oncol,2012,105(3):341-349.
[21]Hasegawa M,Fujimoto M,Takehara K,et al.Pathogenesis of systemic sclerosis:Altered B cell function is the key linking systemic autoimmunity and tissue fibrosis[J].J Dermato Sci,2005,39(1):1-7.
[22]Torres JPD,Pintoplata V,Casanova C,et al.C-Reactive protein levels and survival in patients with moderate to very severe COPD[J].Chest,2008,133(6):1336-1343.
[23]Younis MK,Paul K,Peter J,et al.Brd4 Is essential for IL-1β-induced inflammation in human airway epithelial cells[J].PLo SOne,2014,9(4):e95051.
[24]Baines KJ,Backer V,Gibson PG,et al.Impaired lung function is associated with systemic inflammation and macrophage activation[J].Eur Respiratory J,2014,45(2):557-559.
[25]Jang SS,Kim HG,Lee JS,et al.Melatonin reduces X-ray radiation-induced lung injury in mice by modulating oxidative stress and cytokine expression[J].Intern J Rad Biol,2013,89(2):97-105.
[26]Zhao W,Robbins ME.Inflammation and chronic oxidative stress in radiation-induced late normal tissue injury:therapeutic implications[J].Current Med Chem,2009,16(2):130-143.
[27]Robbins ME,Zhao W.Chronic oxidative stress and radiation-induced late normal tissue injury:a review[J].Int J Radiat Biol,2004,80(4):251-259.
[28]Ghafoori P,Marks LB,Vujaskovic Z,et al.Radiation-induced lung injury.Assessment,management,and prevention[J].Oncology,2008,22(1):52-53.
[29]Tak JK,Park JW.The use of ebselen for radioprotection in cultured cells and mice[J].Free Radical Biol Med,2009,46(8):1177-1185.
[30]Terasaki Y,Ohsawa I,Terasaki M,et al.Hydrogen therapy attenuates irradiation-induced lung damage by reducing oxidative stress[J].Am J Physiol Lung Cell Mol Physiol,2011,301(4):415-426.
[31]Marklund SL.Extracellular superoxide dismutase and other superoxide dismutase isoenzymes in tissues from nine mammalian species[J].Biochem J,1984,222(3):649-655.
[32]Zahid N Rabbani,Mitchell S Anscher,Rodney J Folz,et al.Overexpression of extracellular superoxide dismutase reduces acute radiation induced lung toxicity[J].BMC Cancer,2005,5(1):59.
[33]Thompson HG,Mih JD,Krasieva TB,et al.Epithelial-derived TGF-beta2 modulates basal and wound-healing subepithelial matrix homeostasis[J].Am J Physiol Lung Cell Mol Physiol,2006,291(6):1277-1285.
[34]Khalil N,Bereznay O,Sporn M,et al.Macrophage production of transforming growth factor beta and fibroblast collagen synthesis in chronic pulmonary inflammation[J].J Exp Med,1989,170(3):727-737.
[35]Vujaskovic Z,Groen HJ.TGF-beta,radiation-induced pulmonary injury and lung cancer[J].Intern J Rad Biol,2000,76(4):511-516.
[36]Roberts A B,Russo A,Felici A,et al.Smad3:A Key Player in Pathogenetic Mechanisms Dependent on TGF-β[J].Ann New York Acad Sci,2003,995(1):1-10.
[37]Mata-Greenwood E,Grobe A,Kumar S,et al.Cyclic stretch increases VEGF expression in pulmonary arterial smooth muscle cells via TGF-beta1 and reactive oxygen species:a requirement for NAD(P)H oxidase[J].Am J Physiol Lung Cell Mol Physiol,2005,289(2):288-289.
[2]Martel MK,Ten Haken RK,Hazuka MB,et al.Dose-volume histogram and 3-D treatment planning evaluation of patients with pneumonitis[J].Int J Rad Oncol Biol Phys,1994,28(3):575-581.
[3]Yu B,Wang J,Xu Y,et al.Radiotherapy techniques and radiation pneumonitis:a lot to a little or a little to a lot?[J].Chin J Lung Cancer,2015,18(12):752-757.
[4]Anscher M S,Garst J,Marks L B,et al.Assessing the ability of the antiangiogenic and anticytokine agent thalidomide to modulate radiation-induced lung injury[J].Int J Rad Oncol Biol Phys,2006,66(2):477-482.
[5]Robbins ME,Diz DI.Pathogenic role of the renin-angiotensin system in modulating radiation-induced late effects[J].Int J Rad Oncol Biol Phys,2006,64(1):6-12.
[6]Lee SJ,Yi C,Heo RW,et al.Clarithromycin attenuates radiationinduced lung injury in mice[J].Plos One,2015,10(6):e0131671.
[7]Antonic V,Rabbani ZN,Jackson IL,et al.Subcutaneous administration of bovine superoxide dismutase protects lungs from radiation-induced lung injury[J].Free Radical Res,2015,49(10):1259-1268.
[8]Abdulla S,Salavati A,Saboury B,et al.Quantitative assessment of global lung inflammation following radiation therapy using FDGPET/CT:a pilot study[J].Eur J Nuclear Med Mol Imaging,2014,41(2):350-356.
[9]Tian W,Chen C,Man Y,et al.Ethanol extract mediates rat sex hormone levels by induction of testicular steroidgenic enzymes[J].Pharmaceutical Biol,2016,54(3):481-487.
[10]Xiong Q,Kadota S,Tani T,et al.Antioxidative effects of phenylethanoids from Cistanche deserticola[J].Biol Pharm Bull,1996,19(12):1580-1585.
[11]Wang T,Zhang X,Xie W.Cistanche deserticola Y.C.Ma,"Desert ginseng":a review[J].Am J Chin Med,2012,40(06):1123-1141.
[12]Gu L,Xiong WT,Wang C,et al.Cistanche deserticola decoction alleviates the testicular toxicity induced by hydroxyurea in male mice[J].Asian J Androl,2013,15(6):838-840.
[13]Luo H,Cao R,Wang L,et al.Protective effect of Cistanchis,A on ethanol-induced damage in primary cultured mouse hepatocytes[J].Biomed Pharmacotherapy,2016,83:1071-1079.
[14]Vujaskovic Z,Marks L B,Anscher M S.The physical parameters and molecular events associated with radiation-induced lung toxicity[J].Seminars Rad Oncol,2000,10(4):296-307.
[15]Nan ZD,Zeng KW,Shi SP,et al.Phenylethanoid glycosides with anti-inflammatory activities from the stems of Cistanche deserticola cultured in Tarim desert[J].Fitoterapia,2013,89(1):167-174.
[16]Haimovitzfriedman A,Yang TJ,Thin TH,et al.Imaging Radiotherapy-Induced Apoptosis[J].Rad Res,2012,177(4):467-482.
[17]Ahire V,Mishra KP,Kulkarni GR.On the mechanism of cellular toxicity in breast cancer by ionizing radiation and chemotherapeutic drugs[J].J Environ Pathol Toxicol Oncol,2014,33(1):69-82.
[18]Bakkal BH,Gultekin FA,Guven B,et al.Effect of ozone oxidative preconditioning in preventing early radiation-induced lung injury in rats[J].Brazilian J Med Biol Res,2013,46(9):789-796.
[19]Martin M,Lefaix J,Delanian S.TGF-beta1 and radiation fibrosis:a master switch and a specific therapeutic target?[J].Int J Rad Oncol Biol Phys,2000,47(2):277-290.
[20]Zaidi A,Jelveh S,Mahmood J,et al.Effects of lipopolysaccharide on the response of C57BL/6J mice to whole thorax irradiation[J].Radiotherapy Oncol,2012,105(3):341-349.
[21]Hasegawa M,Fujimoto M,Takehara K,et al.Pathogenesis of systemic sclerosis:Altered B cell function is the key linking systemic autoimmunity and tissue fibrosis[J].J Dermato Sci,2005,39(1):1-7.
[22]Torres JPD,Pintoplata V,Casanova C,et al.C-Reactive protein levels and survival in patients with moderate to very severe COPD[J].Chest,2008,133(6):1336-1343.
[23]Younis MK,Paul K,Peter J,et al.Brd4 Is essential for IL-1β-induced inflammation in human airway epithelial cells[J].PLo SOne,2014,9(4):e95051.
[24]Baines KJ,Backer V,Gibson PG,et al.Impaired lung function is associated with systemic inflammation and macrophage activation[J].Eur Respiratory J,2014,45(2):557-559.
[25]Jang SS,Kim HG,Lee JS,et al.Melatonin reduces X-ray radiation-induced lung injury in mice by modulating oxidative stress and cytokine expression[J].Intern J Rad Biol,2013,89(2):97-105.
[26]Zhao W,Robbins ME.Inflammation and chronic oxidative stress in radiation-induced late normal tissue injury:therapeutic implications[J].Current Med Chem,2009,16(2):130-143.
[27]Robbins ME,Zhao W.Chronic oxidative stress and radiation-induced late normal tissue injury:a review[J].Int J Radiat Biol,2004,80(4):251-259.
[28]Ghafoori P,Marks LB,Vujaskovic Z,et al.Radiation-induced lung injury.Assessment,management,and prevention[J].Oncology,2008,22(1):52-53.
[29]Tak JK,Park JW.The use of ebselen for radioprotection in cultured cells and mice[J].Free Radical Biol Med,2009,46(8):1177-1185.
[30]Terasaki Y,Ohsawa I,Terasaki M,et al.Hydrogen therapy attenuates irradiation-induced lung damage by reducing oxidative stress[J].Am J Physiol Lung Cell Mol Physiol,2011,301(4):415-426.
[31]Marklund SL.Extracellular superoxide dismutase and other superoxide dismutase isoenzymes in tissues from nine mammalian species[J].Biochem J,1984,222(3):649-655.
[32]Zahid N Rabbani,Mitchell S Anscher,Rodney J Folz,et al.Overexpression of extracellular superoxide dismutase reduces acute radiation induced lung toxicity[J].BMC Cancer,2005,5(1):59.
[33]Thompson HG,Mih JD,Krasieva TB,et al.Epithelial-derived TGF-beta2 modulates basal and wound-healing subepithelial matrix homeostasis[J].Am J Physiol Lung Cell Mol Physiol,2006,291(6):1277-1285.
[34]Khalil N,Bereznay O,Sporn M,et al.Macrophage production of transforming growth factor beta and fibroblast collagen synthesis in chronic pulmonary inflammation[J].J Exp Med,1989,170(3):727-737.
[35]Vujaskovic Z,Groen HJ.TGF-beta,radiation-induced pulmonary injury and lung cancer[J].Intern J Rad Biol,2000,76(4):511-516.
[36]Roberts A B,Russo A,Felici A,et al.Smad3:A Key Player in Pathogenetic Mechanisms Dependent on TGF-β[J].Ann New York Acad Sci,2003,995(1):1-10.
[37]Mata-Greenwood E,Grobe A,Kumar S,et al.Cyclic stretch increases VEGF expression in pulmonary arterial smooth muscle cells via TGF-beta1 and reactive oxygen species:a requirement for NAD(P)H oxidase[J].Am J Physiol Lung Cell Mol Physiol,2005,289(2):288-289.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |