同型半胱氨酸经内质网应激介导APOE~(-/-)小鼠心肌细胞凋亡及枸杞多糖的干预作用
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摘要
目的探讨内质网应激(ERS)在同型半胱氨酸(Hcy)介导APOE~(-/-)小鼠心肌细胞凋亡中的作用及枸杞多糖(LBP)的干预效果。方法取36只健康SPF级6周龄雄性APOE~(-/-)小鼠,随机分为对照组(C组)、同型半胱氨酸组(H组)、同型半胱氨酸+枸杞多糖组(HL组),每组12只。C组给予正常饮食,H组、HL组给予高蛋氨酸饮食建立高同型半胱氨酸血症(HHcy)模型,HL组每天上午9:00给予0.1mL LBP溶液灌胃[80mg/(kg·d)-1],C组、H组给予0.1mL生理盐水灌胃。干预24周后处死小鼠,检测血清Hcy浓度,记录心脏脏器系数,TUNEL染色检测各组心肌细胞凋亡差异,免疫组化及蛋白印迹检测ERS相关因子(BIP、ATF6、IRE1、PERK、ERO1-α、CHOP)表达情况。结果与C组相比,H组血清Hcy浓度明显升高,心脏脏器系数增加,心肌细胞凋亡增多(P均<0.05)。ERS相关因子的免疫组化及蛋白印迹结果显示H组较C组的表达量均增多,HL组较H组有所降低(P<0.05)。结论 ERS参与Hcy介导的APOE~(-/-)小鼠心肌细胞凋亡,LBP能减轻ERS而达到保护心肌的作用。
Objective To investigate the effect of endoplasmic reticulum stress(ERS) on apoptosis of APOE~(-/-)mouse myocardial cells induced by homocysteine(Hcy) and the intervention effect of LBP.Methods Thirty-six healthy 6-week-old male APOE~(-/-)mice of SPF grade were randomly divided into control group(C),homocysteine group(H),homocysteine + Lycium barbarum polysaccharide group(HL),12 mice in each group. Mice in C group were given normal diet,and H and HL mice were given high methionine diet to establish a model of hyperhomocysteinemia(HHcy). HL mice were given 0.1 mL LBP solution[(80 mg/(kg·d)-1]at 9:00 a.m. and C and H mice were given 0.1 mL saline. After 24 weeks of intervention,mice were sacrificed,serum Hcy concentration was detected,cardiac organ coefficient was recorded,apoptotic difference of cardiac myocytes was detected by TUNEL staining,and ERS-related factors(BIP,ATF6,IRE1,PERK,ERO1-alpha,CHOP)were detected by immunohistochemistry and Western blotting. Results Compared with mice of C group,serum Hcy concentration in H group mice increased significantly,cardiac organ coefficient increased, and myocardial cell apoptosis also increased.Immunohistochemistry and western blot results indicated that the expression levels of ERS related factors were significantly higher than that of C group,while that of HL group was lower than that of H group,and the difference was statistically significant(P<0.05).Conclusion ERS was involved in the apoptosis of APOE~(-/-)mouse myocardial cells mediated by Hcy,and LBP could alleviate ERS to achieve the effect of protecting myocardium.
Objective To investigate the effect of endoplasmic reticulum stress(ERS) on apoptosis of APOE~(-/-)mouse myocardial cells induced by homocysteine(Hcy) and the intervention effect of LBP.Methods Thirty-six healthy 6-week-old male APOE~(-/-)mice of SPF grade were randomly divided into control group(C),homocysteine group(H),homocysteine + Lycium barbarum polysaccharide group(HL),12 mice in each group. Mice in C group were given normal diet,and H and HL mice were given high methionine diet to establish a model of hyperhomocysteinemia(HHcy). HL mice were given 0.1 mL LBP solution[(80 mg/(kg·d)-1]at 9:00 a.m. and C and H mice were given 0.1 mL saline. After 24 weeks of intervention,mice were sacrificed,serum Hcy concentration was detected,cardiac organ coefficient was recorded,apoptotic difference of cardiac myocytes was detected by TUNEL staining,and ERS-related factors(BIP,ATF6,IRE1,PERK,ERO1-alpha,CHOP)were detected by immunohistochemistry and Western blotting. Results Compared with mice of C group,serum Hcy concentration in H group mice increased significantly,cardiac organ coefficient increased, and myocardial cell apoptosis also increased.Immunohistochemistry and western blot results indicated that the expression levels of ERS related factors were significantly higher than that of C group,while that of HL group was lower than that of H group,and the difference was statistically significant(P<0.05).Conclusion ERS was involved in the apoptosis of APOE~(-/-)mouse myocardial cells mediated by Hcy,and LBP could alleviate ERS to achieve the effect of protecting myocardium.
引文
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[7]李丽芳,杜娟,薛华,等.枸杞多糖对心肌梗死大鼠心肌细胞凋亡的影响[J].宁夏医学杂志,2018,40(2):97-99.
[8]刘楷曦,杨歆然,梁统.枸杞多糖对阿霉素诱导大鼠心肌病的影响[J].医学理论与实践,2014(14):1821-1822.
[9] Hetz C. The unfolded protein response:controlling cell fate decisions under ER stress and beyond[J].Nat Rev Mol Cell Biol,2012,13(2):89-102.
[10] Walter P,Ron D. The unfolded protein response:from stress pathway to homeostatic regulation[J].Science,2011,334(6059):1081-1086.
[11] Pakpoor J,Disanto G,Altmann DR,et al. No evidence for higher risk of cancer in patients with multiple sclerosis taking cladribine[J]. Neurol Neuroimmunol Neuroinflamm,2015,2(6):158.
[12] Zhu J,Liu W,Yu JP,et al. Characterization and hypoglycemic effect of a polysaccharide extracted from the fruit of Lycium barbarum L[J]. Carbohydrate Polymers,2013,98(1):8-16.
[13]马云霞,张俭.慢性心力衰竭患者血清同型半胱氨酸、高密度脂蛋白和高敏肌钙蛋白T水平与心功能的相关性研究[J].宁夏医科大学学报,2018,40(4):424-428.
[14] Zhang ZM,Zhao LY,Zhou YF,et al. Taurine ameliorated homocysteine-induced H9C2 cardiomyocyte apoptosis by modulating endoplasmic reticulum stress[J]. Apoptosis,2017,22(5):647-661.
[15]冯彦.枸杞多糖药理作用与临床应用[J].中国现代药物应用,2016,34(6):278-179.
[16]南亚昀,雍学芳,王礼星.枸杞多糖药理学研究进展[J].天津中医药,2014,31(12):763-765.
[2] Moshal K,Camel C,Kartha G,et al. Cardiac dys-synchronization and arrhythmia in hyperhomocysteinemia[J]. Curr Neurovasc Res,2007,4(4):289-294.
[3] Vizzardi E,Bonadei I,Zanini G,et al. Homocysteine and heart failure:An overview[J]. PRC,2009,4(1):15-21.
[4] Wang XC,Sun WT,Yu CM,et al. ER stress mediates homocysteine-induced endothelial dysfunction:Modulation of IKCa and SKCa channels[J]. Atherosclerosis,2015,242(1):191-198.
[5] Cong GZ,Yan R,Huang H,et al. Involvement of histone methylation in macrophage apoptosis and unstable plaque formation in methionine-induced hyperhomocysteinemic ApoE-/-mice[J]. Life Sciences,2017,173:135-144.
[6]杜海林,杨绍兵,丛广志,等.同型半胱氨酸经PERK磷酸化激活CHOP-ERO1α通路介导心肌细胞凋亡的研究[J].重庆医学,2018,47(5):584-587.
[7]李丽芳,杜娟,薛华,等.枸杞多糖对心肌梗死大鼠心肌细胞凋亡的影响[J].宁夏医学杂志,2018,40(2):97-99.
[8]刘楷曦,杨歆然,梁统.枸杞多糖对阿霉素诱导大鼠心肌病的影响[J].医学理论与实践,2014(14):1821-1822.
[9] Hetz C. The unfolded protein response:controlling cell fate decisions under ER stress and beyond[J].Nat Rev Mol Cell Biol,2012,13(2):89-102.
[10] Walter P,Ron D. The unfolded protein response:from stress pathway to homeostatic regulation[J].Science,2011,334(6059):1081-1086.
[11] Pakpoor J,Disanto G,Altmann DR,et al. No evidence for higher risk of cancer in patients with multiple sclerosis taking cladribine[J]. Neurol Neuroimmunol Neuroinflamm,2015,2(6):158.
[12] Zhu J,Liu W,Yu JP,et al. Characterization and hypoglycemic effect of a polysaccharide extracted from the fruit of Lycium barbarum L[J]. Carbohydrate Polymers,2013,98(1):8-16.
[13]马云霞,张俭.慢性心力衰竭患者血清同型半胱氨酸、高密度脂蛋白和高敏肌钙蛋白T水平与心功能的相关性研究[J].宁夏医科大学学报,2018,40(4):424-428.
[14] Zhang ZM,Zhao LY,Zhou YF,et al. Taurine ameliorated homocysteine-induced H9C2 cardiomyocyte apoptosis by modulating endoplasmic reticulum stress[J]. Apoptosis,2017,22(5):647-661.
[15]冯彦.枸杞多糖药理作用与临床应用[J].中国现代药物应用,2016,34(6):278-179.
[16]南亚昀,雍学芳,王礼星.枸杞多糖药理学研究进展[J].天津中医药,2014,31(12):763-765.
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