虾青素介导Nrf2信号通路减轻大强度运动诱导大鼠心肌细胞的凋亡
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摘要
目的:观察补充虾青素能否介导核因子E2相关因子2(nuclear factor-E2-related factor 2,Nrf2)通路,减少6周大强度训练致大鼠心肌组织氧化应激,降低细胞凋亡水平。方法:7周龄SD雄性大鼠,适应性饲养及训练后随机分为对照组(C组,10只)、大强度训练组(HT组,15只)、虾青素+大强度训练组(HTA组,15只),大强度训练持续6周。训练期间,HTA组每天灌胃1次虾青素,灌胃剂量为20 mg/kg/d,C组和HT组分别灌胃同体积的大豆油。实验结束24 h后取血清测试血清肌钙蛋白I(Cardiac Troponin I,cTNI),取心肌测定凋亡指数、B细胞淋巴瘤/白血病基因2(B cell lymphoma/leukemia 2,Bcl2)、Bcl2相关x蛋白(Bcl2 associated x protein,Bax)、Nrf2、血红素加氧酶-1(heme oxygenase-1,HO-1)表达,同时测定心肌总抗氧化能力(Total antioxidative capacity,T-AOC)、超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(Malonaldehyde,MDA)浓度。结果:经6周大强度训练,与C组比较,HT组大鼠血清cTNI、心肌凋亡水平、Bax表达、MDA浓度极显著增强(p<0.01),Bcl2/Bax、HO-1表达量、SOD、T-AOC活性极显著下降(p<0.01)。而通过6周的虾青素干预,与HT组比较,HTA组血清cTNI、心肌MDA浓度、心肌组织细胞的凋亡水平、Bax表达显著下降(p<0.05),Nrf2和HO-1表达量、Bcl2/Bax、SOD和T-AOC活性显著升高(p<0.05)。除Bcl2和HO-1外,其余指标与C组相比仍有显著性差异(p<0.05)。结论:补充虾青素可以介导Nrf2通路,上调Nrf2、HO-1的蛋白表达,提高SOD和T-AOC抗氧化酶的活性,进而有效降低6周大强度运动训练引发的大鼠心脏组织氧化应激水平和心肌细胞凋亡水平,保护心脏组织结构和功能正常。
Objective:By supplementing the astaxanthin,the effects if astaxanthin could mediate Nrf2 and reduce myocardium oxidative injury in rats after high intensity training of 6 weeks were investigated in the research. Methods:7-week SD male rats were divided into 3 groups randomly:control group(C group,n=10),high intensity training group(HT group,n=15),astaxanthin and high intensity training group(HTA group,n=15). The rats in HTA group were intragastrically daily administered astaxanthin 20 mg/kg and were intragastrically administered equal amount of soybean oil in HT group during the training day.The serum cardiac troponin I(cTNI),myocardial apoptosis index,the expression of myocardial B cell lymphoma/leukemia 2(Bcl2),Bcl2 associated x protein(Bax),nuclear factor-E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),myocardial malonaldehyde(MDA),superoxide dismutase(SOD)and total antioxidative capacity(T-AOC)activity were detected at 24 h after the last training.Results:After 6-week of high intensity training,compared with C group,the serum cTNI,myocardial apoptosis index,the Bax expression and myocardial MDA were significantly higher in HT group(p<0.01). The Bcl2/Bax,the expression of HO-1,SOD and T-AOC activity were significantly lower(p<0.01). After the intervention of 6-week astaxanthin intragastrical administration,in HTA group,the serum cTNI,myocardial MDA,the myocardial apoptosis index,the Bax expression were significantly lower than HT group(p<0.05). Finally,Bcl2/Bax,SOD,T-AOC activity,the Nrf2 expression and HO-1 were significantly improved(p<0.05). Compared with C group,all these indexes increased significantly besides Bcl2 and HO-1(p<0.05). Conclusion:These results demonstrate that supplement of astaxanthin can mediate Nrf2 signaling pathway,and elevate the expression of Nrf2 and HO-1.It can also effectively increase the activity of SOD and T-AOC,after that the astaxanthin is helpful to alleviate the myocardial oxidative level and myocardial apoptosis which are caused by 6-week high intensity training. Finally,the structure and function of heart tissue are back to normal.
Objective:By supplementing the astaxanthin,the effects if astaxanthin could mediate Nrf2 and reduce myocardium oxidative injury in rats after high intensity training of 6 weeks were investigated in the research. Methods:7-week SD male rats were divided into 3 groups randomly:control group(C group,n=10),high intensity training group(HT group,n=15),astaxanthin and high intensity training group(HTA group,n=15). The rats in HTA group were intragastrically daily administered astaxanthin 20 mg/kg and were intragastrically administered equal amount of soybean oil in HT group during the training day.The serum cardiac troponin I(cTNI),myocardial apoptosis index,the expression of myocardial B cell lymphoma/leukemia 2(Bcl2),Bcl2 associated x protein(Bax),nuclear factor-E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),myocardial malonaldehyde(MDA),superoxide dismutase(SOD)and total antioxidative capacity(T-AOC)activity were detected at 24 h after the last training.Results:After 6-week of high intensity training,compared with C group,the serum cTNI,myocardial apoptosis index,the Bax expression and myocardial MDA were significantly higher in HT group(p<0.01). The Bcl2/Bax,the expression of HO-1,SOD and T-AOC activity were significantly lower(p<0.01). After the intervention of 6-week astaxanthin intragastrical administration,in HTA group,the serum cTNI,myocardial MDA,the myocardial apoptosis index,the Bax expression were significantly lower than HT group(p<0.05). Finally,Bcl2/Bax,SOD,T-AOC activity,the Nrf2 expression and HO-1 were significantly improved(p<0.05). Compared with C group,all these indexes increased significantly besides Bcl2 and HO-1(p<0.05). Conclusion:These results demonstrate that supplement of astaxanthin can mediate Nrf2 signaling pathway,and elevate the expression of Nrf2 and HO-1.It can also effectively increase the activity of SOD and T-AOC,after that the astaxanthin is helpful to alleviate the myocardial oxidative level and myocardial apoptosis which are caused by 6-week high intensity training. Finally,the structure and function of heart tissue are back to normal.
引文
[1]周庆新,刘婷婷,杨鲁. 虾青素的来源、生物功效及吸收代谢研究进展[J]. 食品研究与开发,2017,38(16):214-219.
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[5]Vidotto C,Tschan H,Atamaniuk J,et al. Responses of N-terminal pro-brain natriuretic peptide(NT-proBNP)and cardiac troponin I(cTnI)to competitive endurance exercise in recreational athletes[J]. International Journal of Sports Medicine,2005,26(8):645-650.
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[9]倪晓锋,于海宁,王姗姗,等. 虾青素保护 PC-3 细胞免受 H2O2氧化应激的作用机制[J]. 食品科学,2018,39(9):154-162.
[10]郑宁. 补充天然虾青素对递增负荷训练大鼠骨骼肌自由基代谢的影响[J].成都体育学院学报,2014,40(2):75-79.
[11]周海涛,曹建民,郭娴,等. 虾青素对大鼠运动性心肌损伤的保护作用[J].天然产物研究与开发,2016,28:1150-1155.
[12]曹敏敏. SPAG9在宫颈腺癌中的表达及其临床意义[D]. 济南:山东大学,2014.
[13]庞立杰. 大鼠过度训练后血清cTnI、CK-MB和TNF-α的变化及复方丹参的干预效应[D]. 太原:山西大学,2007.
[14]王凤娟. 实验性心肌损伤大鼠血清心肌肌钙蛋白Ⅰ变化的比较研究[D].上海:上海体育学院,2010.
[15]路巍. 红景天苷通过Keap1-Nrf2-ARE信号通路对大鼠力竭心脏氧化应激的调控作用[D].石家庄:河北医科大学,2016.
[16]潘珊珊,孙晓娟,尤培建. 过度训练对心肌和骨骼肌缺血缺氧改变的形态学研究[J]. 体育科学,2005,25(12):49-52.
[17]王凤娟. 实验性心肌损伤大鼠血清心肌肌钙蛋白I变化的比较研究[D].上海:上海体育学院,2010.
[18]杜洁,李增明. 人参黄连对力竭运动大鼠血清肌钙蛋白T、脑钠肽以及心脏的可能影响实验研究[J]. 广州体育学院学报,2016,36(6):97-99.
[19]王瑜娟. 不同强度力竭运动对大鼠心肌细胞凋亡的影响[D].长沙:湖南师范大学,2005.
[20]Ishii T,Itoh K,Takahashi S,et al. Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages[J]. The Journal of Biological Chemistry,2000,275(21):16023-16029.
[21]Reuland D J,Khademi S,Castle C J,et al. Upregulation of phase II enzymes through phytochemical activation of Nrf2 protects cardiomyocytes against oxidant stress[J]. Free Radical Biology & Medicine,2013,56:102-111.
[22]陶天琪,王晓礽,徐菲菲,等. MR-1通过抑制PERK/Nrf2途径减轻缺氧/复氧诱导的心肌细胞凋亡[J]. 中国病理生理杂志,2014,30(2):193-202.
[23]杨芳,吴泽幼,包思,等. 松果菊苷通过调节Nrf2表达减轻H2O2致心肌细胞损伤的实验研究[J]. 中国中医急症,2017,26(12):2078-2082.
[24]Satoh T,Mckercher S R,Lipton S A. Reprint of:Nrf2/ARE-mediated antioxidant actions of pro-electrophilic drugs[J]. Free Radical Biology & Medicine,2014,66(2):45-57.
[25]Tsai C Y,Wang C C,Lai T Y,et al. Antioxidant effects of diallyl trisulfide on high glucose-induced apoptosis are mediated by the PI3K/Akt-dependent activation of Nrf2 in cardiomyocytes[J]. International Journal of Cardiology,2013,168(2):1286-1297.
[26]Chan K C,Mong M C,Yin M C. Antioxidative and anti-inflammatory neuroprotective effects of astaxanthin and canthaxanthin in nerve growth factor differentiated PC12 cells[J].Journal of Food Science,2009,74(7):H225-231.
[27]Song X,Wang B,Lin S,et al. Astaxanthin inhibits apoptosis in alveolar epithelial cells type II in vivo and in vitro through the ROS-dependent mitochondrial signalling pathway[J]. Journal of Cellular and Molecular Medicine,2014,18(11):2198-2212.
[2]Shen M,Chen K,Lu J,et al. Protective effect of astaxanthin on liver fibrosis through modulation of TGF-beta1 expression and autophagy[J]. Mediators of Inflammation,2014,2014:1-14.
[3]Li J,Xia Y,Liu T,et al. Protective effects of astaxanthin on ConA-induced autoimmune hepatitis by the JNK/p-JNK pathway-mediated inhibition of autophagy and apoptosis[J]. Plos One,2015,10(3):e0120440.
[4]Shave R,Ross P,Low D,et al. Cardiac troponin I is released following high-intensity short-duration exercise in healthy humans[J]. International Journal of Cardiology,2010,145(2):337-339.
[5]Vidotto C,Tschan H,Atamaniuk J,et al. Responses of N-terminal pro-brain natriuretic peptide(NT-proBNP)and cardiac troponin I(cTnI)to competitive endurance exercise in recreational athletes[J]. International Journal of Sports Medicine,2005,26(8):645-650.
[6]Sinha M K,Gaze D C,Tippins J R,et al. Ischemia modified albumin is a sensitive marker of myocardial ischemia after percutaneous coronary intervention[J]. Circulation,2003,107(19):2403-2405.
[7]宋晓冬,王美蓉,张瑾锦,等. 虾青素对大鼠肝癌CBRH-7919细胞骨架和nm23蛋白的影响[J]. 滨州医学院学报,2010,33(5):321-324.
[8]葛晓平,汪云开. 虾青素对过氧化氢损伤心肌细胞的保护作用[J]. 中华危重症医学杂志(电子版),2011,4(1):35-38.
[9]倪晓锋,于海宁,王姗姗,等. 虾青素保护 PC-3 细胞免受 H2O2氧化应激的作用机制[J]. 食品科学,2018,39(9):154-162.
[10]郑宁. 补充天然虾青素对递增负荷训练大鼠骨骼肌自由基代谢的影响[J].成都体育学院学报,2014,40(2):75-79.
[11]周海涛,曹建民,郭娴,等. 虾青素对大鼠运动性心肌损伤的保护作用[J].天然产物研究与开发,2016,28:1150-1155.
[12]曹敏敏. SPAG9在宫颈腺癌中的表达及其临床意义[D]. 济南:山东大学,2014.
[13]庞立杰. 大鼠过度训练后血清cTnI、CK-MB和TNF-α的变化及复方丹参的干预效应[D]. 太原:山西大学,2007.
[14]王凤娟. 实验性心肌损伤大鼠血清心肌肌钙蛋白Ⅰ变化的比较研究[D].上海:上海体育学院,2010.
[15]路巍. 红景天苷通过Keap1-Nrf2-ARE信号通路对大鼠力竭心脏氧化应激的调控作用[D].石家庄:河北医科大学,2016.
[16]潘珊珊,孙晓娟,尤培建. 过度训练对心肌和骨骼肌缺血缺氧改变的形态学研究[J]. 体育科学,2005,25(12):49-52.
[17]王凤娟. 实验性心肌损伤大鼠血清心肌肌钙蛋白I变化的比较研究[D].上海:上海体育学院,2010.
[18]杜洁,李增明. 人参黄连对力竭运动大鼠血清肌钙蛋白T、脑钠肽以及心脏的可能影响实验研究[J]. 广州体育学院学报,2016,36(6):97-99.
[19]王瑜娟. 不同强度力竭运动对大鼠心肌细胞凋亡的影响[D].长沙:湖南师范大学,2005.
[20]Ishii T,Itoh K,Takahashi S,et al. Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages[J]. The Journal of Biological Chemistry,2000,275(21):16023-16029.
[21]Reuland D J,Khademi S,Castle C J,et al. Upregulation of phase II enzymes through phytochemical activation of Nrf2 protects cardiomyocytes against oxidant stress[J]. Free Radical Biology & Medicine,2013,56:102-111.
[22]陶天琪,王晓礽,徐菲菲,等. MR-1通过抑制PERK/Nrf2途径减轻缺氧/复氧诱导的心肌细胞凋亡[J]. 中国病理生理杂志,2014,30(2):193-202.
[23]杨芳,吴泽幼,包思,等. 松果菊苷通过调节Nrf2表达减轻H2O2致心肌细胞损伤的实验研究[J]. 中国中医急症,2017,26(12):2078-2082.
[24]Satoh T,Mckercher S R,Lipton S A. Reprint of:Nrf2/ARE-mediated antioxidant actions of pro-electrophilic drugs[J]. Free Radical Biology & Medicine,2014,66(2):45-57.
[25]Tsai C Y,Wang C C,Lai T Y,et al. Antioxidant effects of diallyl trisulfide on high glucose-induced apoptosis are mediated by the PI3K/Akt-dependent activation of Nrf2 in cardiomyocytes[J]. International Journal of Cardiology,2013,168(2):1286-1297.
[26]Chan K C,Mong M C,Yin M C. Antioxidative and anti-inflammatory neuroprotective effects of astaxanthin and canthaxanthin in nerve growth factor differentiated PC12 cells[J].Journal of Food Science,2009,74(7):H225-231.
[27]Song X,Wang B,Lin S,et al. Astaxanthin inhibits apoptosis in alveolar epithelial cells type II in vivo and in vitro through the ROS-dependent mitochondrial signalling pathway[J]. Journal of Cellular and Molecular Medicine,2014,18(11):2198-2212.