当归多糖对缺氧-复氧损害导致的H9c2心肌细胞能量代谢障碍的保护作用
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摘要
目的:探索当归多糖(ASP)对缺氧-复氧损害导致的H9c2心肌细胞能量代谢障碍的保护作用。方法:MTT检测ASP对H9c2细胞增殖的影响;在体外建造ASP干预和非干预下的缺氧-复氧模型;观察细胞形态;流式检测细胞凋亡;线粒体膜电位(MMP)检测试剂盒检测细胞线粒体膜电位变化;试剂盒检测心肌细胞培养上清液中LDH和心肌细胞中Na~+-K~+ATP酶和Ca~(2+)-Mg~(2+)ATP酶以及线粒体呼吸酶链复合物Ⅰ、Ⅳ活性。结果:ASP浓度-时间依赖地抑制H9c2细胞增殖;缺氧-复氧诱导H9c2细胞凋亡、MMP减小,同时造成H9c2细胞上清中LDH上升,细胞中Na~+-K~+ATP酶、Ca~(2+)-Mg~(2+)ATP酶以及线粒体呼吸酶链复合物Ⅰ、Ⅳ活性降低; ASP可浓度依赖的抑制细胞凋亡,提高MMP,降低上清中LDH,增加Na~+-K~+ATP酶、Ca~(2+)-Mg~(2+)ATP酶以及线粒体呼吸酶链复合物Ⅰ、Ⅳ活性。结论:ASP可通过改善缺氧-复氧造成的心肌细胞能量代谢障碍,保护心肌细胞免受缺氧-复氧的损伤。
Objective: To explore the protective effect of Angelica polysaccharide (ASP) on energy metabolism disorder of H9c2 myocardial cells induced by hypoxia-reoxygenation damage. Methods: MTT was used to detect the effect of polysaccharides from Angelica sinensis on the proliferation of H9c2 cells. The hypoxia-reoxygenation model of Angelica sinensis polysaccharide intervention and non-intervention was constructed in vitro; cell morphology was observed; apoptosis was detected by flow cytometry; mitochondrial membrane potential detection kit was used to detect cells. Mitochondrial membrane potential changes; kits were used to detect the activity of Na~+-K~+ ATPase and Ca~(2+)-Mg~(2+)ATPase and mitochondrial respiratory enzyme chain complex Ⅰ and Ⅳ in LDH and cardiomyocytes in culture supernatant of cardiomyocytes. Results:Angelica polysaccharide inhibited the proliferation of H9c2 cells in a concentration-dependent manner. Hypoxia-reoxygenation induced apoptosis of H9c2 cells,decreased mitochondrial membrane potential,and increased LDH in H9c2 cell supernatant. Na~+-K~+ ATPase and Ca~(2+)-Mg~(2+)ATPase in cells and mitochondrial respiratory enzyme chain complex Ⅰ,Ⅳ activity decreased; Angelica polysaccharide can inhibit cell apoptosis in a concentration-dependent manner,increase mitochondrial membrane potential,reduce LDH in supernatant,increase Na~+-K~+ ATPase,Ca~(2+)-Mg~(2+)ATPase and mitochondrial respiratory enzyme chain complex Ⅰ,Ⅳ activity.Conclusion: Angelica polysaccharide can protect cardiomyocytes from hypoxia-reoxygenation injury by improving myocardial cell energy metabolism disorder caused by hypoxia-reoxygenation.
Objective: To explore the protective effect of Angelica polysaccharide (ASP) on energy metabolism disorder of H9c2 myocardial cells induced by hypoxia-reoxygenation damage. Methods: MTT was used to detect the effect of polysaccharides from Angelica sinensis on the proliferation of H9c2 cells. The hypoxia-reoxygenation model of Angelica sinensis polysaccharide intervention and non-intervention was constructed in vitro; cell morphology was observed; apoptosis was detected by flow cytometry; mitochondrial membrane potential detection kit was used to detect cells. Mitochondrial membrane potential changes; kits were used to detect the activity of Na~+-K~+ ATPase and Ca~(2+)-Mg~(2+)ATPase and mitochondrial respiratory enzyme chain complex Ⅰ and Ⅳ in LDH and cardiomyocytes in culture supernatant of cardiomyocytes. Results:Angelica polysaccharide inhibited the proliferation of H9c2 cells in a concentration-dependent manner. Hypoxia-reoxygenation induced apoptosis of H9c2 cells,decreased mitochondrial membrane potential,and increased LDH in H9c2 cell supernatant. Na~+-K~+ ATPase and Ca~(2+)-Mg~(2+)ATPase in cells and mitochondrial respiratory enzyme chain complex Ⅰ,Ⅳ activity decreased; Angelica polysaccharide can inhibit cell apoptosis in a concentration-dependent manner,increase mitochondrial membrane potential,reduce LDH in supernatant,increase Na~+-K~+ ATPase,Ca~(2+)-Mg~(2+)ATPase and mitochondrial respiratory enzyme chain complex Ⅰ,Ⅳ activity.Conclusion: Angelica polysaccharide can protect cardiomyocytes from hypoxia-reoxygenation injury by improving myocardial cell energy metabolism disorder caused by hypoxia-reoxygenation.
引文
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[2]吴晓燕,苗琳,郑蕊,等.心肌缺血再灌注损伤的研究进展[J].中国临床药理学杂志,2016,32(11):1043~1045.
[3]Lalota J.Molecular mechanism of ischemia-reperfusion injury after myocardial infarction and its possible targeted treatment[J].International Journal of Cardiology,2016,220:571~572.
[4]丁士骜,梅举.线粒体与心肌缺血/再灌注损伤的研究进展[J].医学综述,2016,22(1):37~41.
[5]林荣,林琳,贾兴泽,等.心肌缺血再灌注损伤后对线粒体功能的影响和能量代谢的变化研究[J].贵州医药,2016,40(6):589~590.
[6]张尚智,朱田田,晋玲,等.当归单体成分与复合制剂功效研究进展[J].中国中医药信息杂志,2016,23(12):128~132.
[7]Daz A,Humeres C,Gonxlez V,et al.Insulin/NFκB protects against ischemia-induced necrotic cardiomyocyte death[J].Biochemical and Biophysical Research Communications,2015,467(2):451~457.
[8]张海红,王树,王金,等.当归多糖抗氧化及神经保护作用研究进展[J].神经药理学报,2014,4(2):39~45.
[9]陈福晖,刘达兴,容松.心肌缺血再灌注损伤发生机制的研究进展[J].安徽医药,2017,21(12):2145~2148.
[10]吕宝北,赵鹏翔,张鑫,等.线粒体呼吸链复合物I结构和功能的研究进展[J].现代生物医学进展,2018,2:356~359,38.