青蒿总黄酮联合有氧间歇运动训练抑制肝纤维化小鼠模型炎症反应
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摘要
利用青蒿总黄酮联合有氧间歇运动训练处理肝纤维化小鼠模型,检测肝纤维化小鼠模型中肝纤维化程度及炎症相关因子的变化,探究青蒿总黄酮及有氧间歇运动训练在治疗肝纤维化过程中的作用。取雄性Balb/c小鼠40只,刀豆蛋白A尾静脉注射成模后随机分为模型组(Model组)、黄酮组(FLA组)、运动组(TM组),联合组(FTM组)。用Masson染色检测各组小鼠肝脏纤维化程度,实时荧光定量PCR检测白细胞介素-1β(IL-1β)、IL-10、IL-6以及肿瘤坏死因子α(TNF-α)的表达;用ELISA方法检测IL-1β、IL-10以及TNF-α的表达。结果显示,与模型组相比,运动组及黄酮组肝纤维化程度显著降低,联合组纤维化程度极显著减少;黄酮组和运动组中IL-1β、IL-6、TNF-α的转录水平显著低于模型组,联合组中含量极显著降低,IL-10呈现相反的趋势;血清中IL-1β、TNF-α的含量在黄酮组和运动组中亦显著下降,联合组中含量极显著下降,而IL-10含量则相反。说明青蒿总黄酮及运动训练联合使用能够有效降低小鼠模型中肝纤维化程度,降低肝脏炎症反应,提示其能够阻滞肝纤维化向肝癌的发展。
To study the mechanisms of flavonoids in artemisia combined with aerobic interval training against inflammation in liver fibrosis mice,forty health male Balb/c mice were treated with ConA to develop hepatic fibrosis models.Then,mice were randomly divided into model,flavonoids,treadmill and combined group.Masson staining was carried out to detect the degree of hepatic fibrosis among the groups.The hepatic expression levels of inflammatory factors,such as interleukin-1β(IL-1β),interleukin-10(IL-10),interleukin-6(IL-6),and tumor necrosis factorα(TNF-α),were checked by real-time polymerase chain reaction(real-time PCR).The differences of serum IL-1β,IL-10,and TNF-αwere detected by enzyme linked immunosorbent assay(ELISA).The degree of hepatic fibrosis was deteriorated in the model group,but significantly relieved in the aerobic interval training group(TM),flavonoids group(FLA),and combined group(FTM).In addition,the transcription levels of inflammatory factors were significantly declined in the TM,FLA,and FTM groups,while the expression level of IL-10 was increased significantly in the TM,FLA,and FTM groups.Finally,the serum levels of IL-1βand TNF-αwere significantly decreased in the TM,FLA,and FTM groups,but serum content of IL-10 displayed a reversal trend.Both the treadmill training and flavonoids can ameliorate the hepatic fibrosis,furthermore,the combination of them showed better effect,which may be related with the enhanced anti-inflammation ability.
To study the mechanisms of flavonoids in artemisia combined with aerobic interval training against inflammation in liver fibrosis mice,forty health male Balb/c mice were treated with ConA to develop hepatic fibrosis models.Then,mice were randomly divided into model,flavonoids,treadmill and combined group.Masson staining was carried out to detect the degree of hepatic fibrosis among the groups.The hepatic expression levels of inflammatory factors,such as interleukin-1β(IL-1β),interleukin-10(IL-10),interleukin-6(IL-6),and tumor necrosis factorα(TNF-α),were checked by real-time polymerase chain reaction(real-time PCR).The differences of serum IL-1β,IL-10,and TNF-αwere detected by enzyme linked immunosorbent assay(ELISA).The degree of hepatic fibrosis was deteriorated in the model group,but significantly relieved in the aerobic interval training group(TM),flavonoids group(FLA),and combined group(FTM).In addition,the transcription levels of inflammatory factors were significantly declined in the TM,FLA,and FTM groups,while the expression level of IL-10 was increased significantly in the TM,FLA,and FTM groups.Finally,the serum levels of IL-1βand TNF-αwere significantly decreased in the TM,FLA,and FTM groups,but serum content of IL-10 displayed a reversal trend.Both the treadmill training and flavonoids can ameliorate the hepatic fibrosis,furthermore,the combination of them showed better effect,which may be related with the enhanced anti-inflammation ability.
引文
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[24]胡茂志,赵维芯,康喜龙,等.鼠源炎症因子流式蛋白定量试剂盒的优化及初步应用[J].动物医学进展,2017,38(5):17-21.
[25] Koyama Y,Brenner D A.Liver inflammation and fibrosis[J].Int J cancer,2017,127(1):55-64.
[26] Angulo P,Keach J C,Batts K P,et al.Independent predictors of liver fibrosis in patients with nonalcoholic steatohepatitis[J].Hepatol,1999,30(6):1356-1362.
[27] Bi Y,Mukhopadhyay D,Drinane M,et al.Endocytosis of collagen by hepatic stellate cells regulates extracellular matrix dynamics[J].Am J Physiol-Cell P,2014,307(7):C622-633.
[28] Affo S,Yu L,Schwabe R F.The role of cancer-associated fibroblasts and fibrosis in liver cancer[J].Annu Rev Pathol,2016,12(1):153-186.
[29]陈飞虎,袁丽萍,钟明媚,等.鬼针草总黄酮抗大鼠肝纤维化的实验研究[J].中国临床药理学与治疗学,2006,11(12):1369-1374.
[30]石晓,卓菊.赶黄草总黄酮抗大鼠酒精性肝纤维化作用的实验研究[J].中药材,2015,38(7):1485-1487.
[31]吴景华,郭佳培,曹青,等.青蒿素通过抑制IL-17/IL-17R表达诱导肝癌细胞凋亡的机制研究[J].现代预防医学,2017,44(4):697-700.
[2] Chen W,Zheng R,Baade P D,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[3] Chen W.Cancer statistics:updated cancer burden in China[J].Chin J Cancer Res,2015,27(1):1.
[4]中华人民共和国卫生和计划生育委员会医政医管局.原发性肝癌诊疗规范(2017年版)[J].传染病信息,2017,16(3):705-720.
[5] Ferlay J,Soerjomataram I,Dikshit R,et al.Cancer incidence and mortality worldwide:sources,methods and major patterns in GLOBOCAN 2012[J].Int J Cancer,2015,136(5):E359.
[6] Unalparida A,Ruhl C E.Liver fibrosis scores predict liver disease mortality in the United States population[J].Hepatology,2017,66(1):84-95.
[7] Alves C R,da Cunha T F,da Paixao N A,et al.Aerobic exercise training as therapy for cardiac and cancer cachexia[J].Life Sci,2015,125:9-14.
[8] Zhan L,Huang C,Meng X M,et al.Hypoxia-inducible factor-1alpha in hepatic fibrosis:A promising therapeutic target[J].Biochimie,2015,108(C):1-7.
[9] Roychowdhury S,Chiang D J,McMullen M R,et al.Moderate,chronic ethanol feeding exacerbates carbon-tetrachloride-induced hepatic fibrosis via hepatocyte-specific hypoxia inducible factor 1alpha[J].Pharmacol Res Perspect,2014,2(5):e00061.
[10] Eltzschig H K,Carmeliet P.Hypoxia and inflammation[J].New Engl J Med,2011,364(7):656-665.
[11] Aggarwal H,Nair J,Sharma P,et al.Aegle marmelos differentially affects hepatic markers of glycolysis,insulin signalling pathway,hypoxia,and inflammation in HepG2 cells grown in fructose versus glucose-rich environment[J].Mol Cell Biochem,2017,https://doi.org/10.1007/s11010-017-3108-8.
[12] Feng J,Wang Q S,Chiang A,et al.The effects of sleep hypoxia on coagulant factors and hepatic inflammation in emphysematous rats[J].PLoS One,2010,5(10):e13201.
[13] Dimeo F,Rumberger B G,Keul J.Aerobic exercise as therapy for cancer fatigue[J].Med Sci Sports Exerc,1998,30(4):475-478.
[14] Crevenna R,Schmidinger M,Keilani M,et al.Aerobic exercise as additive palliative treatment for a patient with advanced hepatocellular cancer[J].Wien Med Wochenschr,2003,153(9-10):237-240.
[15] Klika R J,Callahan K E,Drum S N.Individualized 12-week exercise training programs enhance aerobic capacity of cancer survivors[J].Phys Sportsmed,2009,37(3):68-77.
[16] Jones L W,Viglianti B L,Tashjian J A,et al.Effect of aerobic exercise on tumor physiology in an animal model of human breast cancer[J].J Appl Physiol,2010,108(2):343-348.
[17] Ashcraft K A,Peace R M,Betof A S,et al.Efficacy and mechanisms of aerobic exercise on cancer initiation,progression,and metastasis:A critical systematic review of in vivo preclinical data[J].Cancer Res,2016,76(14):4032-4050.
[18] Brown J C,Zemel B S,Troxel A B,et al.Dose-response effects of aerobic exercise on body composition among colon cancer survivors:a randomised controlled trial[J].Br J Cancer,2017,117(11):1614-1620
[19] Kapravelou G,Martinez R,Nebot E,et al.The combined intervention with germinated Vigna radiata and aerobic interval training protocol is an effective strategy for the treatment of non-alcoholic fatty liver disease(NAFLD)and other alterations related to the metabolic syndrome in Zucker rats[J].Nutrients,2017,9(7):774-797
[20]罗伟生,龚受基,梁荣感,等.荔枝核黄酮类化合物体外抗流感病毒作用的研究[J].中国中药杂志,2006,31(16):1379-1380.
[21]谢婧雯,陈妍,贾蔓菁,等.欧芹根总黄酮对非酒精性脂肪性肝病大鼠的治疗作用及机制研究[J].中国药业,2016,25(20):29-32.
[22]张永昕,俞发.枳椇子总黄酮治疗酒精性肝病及其作用机制研究[J].中药材,2010,33(11):1782-1785.
[23] Batatinha H,Lima E A,Teixeira A A S,et al.Association between aerobic exercise and rosiglitazone avoided the NAFLD and liver inflammation exacerbated in PPAR-αknockout mice[J].J Cell Physiol,2017,232(5):1008-1019.
[24]胡茂志,赵维芯,康喜龙,等.鼠源炎症因子流式蛋白定量试剂盒的优化及初步应用[J].动物医学进展,2017,38(5):17-21.
[25] Koyama Y,Brenner D A.Liver inflammation and fibrosis[J].Int J cancer,2017,127(1):55-64.
[26] Angulo P,Keach J C,Batts K P,et al.Independent predictors of liver fibrosis in patients with nonalcoholic steatohepatitis[J].Hepatol,1999,30(6):1356-1362.
[27] Bi Y,Mukhopadhyay D,Drinane M,et al.Endocytosis of collagen by hepatic stellate cells regulates extracellular matrix dynamics[J].Am J Physiol-Cell P,2014,307(7):C622-633.
[28] Affo S,Yu L,Schwabe R F.The role of cancer-associated fibroblasts and fibrosis in liver cancer[J].Annu Rev Pathol,2016,12(1):153-186.
[29]陈飞虎,袁丽萍,钟明媚,等.鬼针草总黄酮抗大鼠肝纤维化的实验研究[J].中国临床药理学与治疗学,2006,11(12):1369-1374.
[30]石晓,卓菊.赶黄草总黄酮抗大鼠酒精性肝纤维化作用的实验研究[J].中药材,2015,38(7):1485-1487.
[31]吴景华,郭佳培,曹青,等.青蒿素通过抑制IL-17/IL-17R表达诱导肝癌细胞凋亡的机制研究[J].现代预防医学,2017,44(4):697-700.