刺五加叶皂苷对乳鼠心肌细胞氧化应激损伤的保护作用

Acanthopanax senticosus saponins ameliorates oxidative damage induced by hydrogen peroxide in neonatal rat cardiomyocytes

目的:探讨刺五加叶皂苷(ASS)对乳鼠心肌细胞氧化应激损伤的保护作用。方法:采用过氧化氢(H2O2)诱导W istar大鼠乳鼠心肌细胞氧化损伤,给予ASS 600,300 mg.L-1,通过细胞形态学变化、细胞存活率、培养液乳酸脱氢酶(LDH)活性及细胞内丙二醛(MDA)含量来反映氧化损伤程度;同时通过检测细胞内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及谷胱甘肽过氧化物酶(GSH-Px)的活性及还原型谷胱甘肽(GSH)含量,探讨ASS对氧化应激损伤的影响。结果:50,100,200μmol.L-1的H2O2均能够诱导心肌细胞发生氧化应激性损伤:心肌细胞存活率显著降低(P<0.01或P<0.001),细胞培养液中LDH的活性及细胞内MDA含量明显增加(P<0.05,P<0.01或P<0.001);同时心肌细胞的形态学发生损伤性改变。而且,随着H2O2浓度的增加,心肌细胞的氧化损伤程度越严重。ASS 600 mg.L-1能减轻H2O2(100μmol.L-1)诱导的氧化应激损伤:LDH的活性(1 687.40±97.51)U.mL-1及细胞内MDA含量(16.50±2.66)nmol.mg-1,差异有统计学意义(P<0.01和P<0.05)。还可以增加细胞内GSH含量(8.91±1.06)μmol.mg-1及GSH-Px,SOD,CAT活性(845.87±63.76)mU.mg-1,(89.55±6.93),(93.07±10.40)U.mg-1,差异显著(P<0.05)。结论:ASS对100μmol.L-1H2O2所致的氧化应激性损伤有明显的保护作用,其机制可能与其抑制脂质过氧化反应,增强心肌细胞抗氧化能力有关。

Objective： To investigate the effect of Acanthopanax senticosus saponins （ASS） on oxidative damage induced by hydrogen peroxide （ H2O2 ） in eardiomyocytes. Method ： The cardiomyocytes were induced to oxidative damaged by exposed to H2O2. We evaluated the level of oxidative injury through morphology change, the survival rate, the activity of lactate dehydrogenase （LDH） and the content of cellular malondialdehyde （MDA）. The activities of superoxide dismutase （SOD）, glutathione peroxidase （GSH- Px）, catalase （CAT） and the content of reductive glutathione （GSH） of cardiomyocytes were also determinted. Result： When the cardiomyocytes were exposed to H2O2 （50, 100,200 μmol·L^-1 ） for deigned time, the percentage of survival cells was down significantly （ P 〈 0.01 or P 〈 0. 001 ）, and the activity of LDH and the content of MDA were increased markedly （ P 〈 0.05, P 〈 0. 01 or P 〈 0. 001 ）. These results show that the cells were subjected to oxidative damage. Treatment with ASS （600 mg ·L^-1 ） prior to H2O2 exposure could increase cell viability significantly, lessen cardiomyocyte morphological damaged change, and decrease LDH activity （ 1 687.40 ±97.51 ） U ·mL^-1 in media and cellular MDA content （ 16. 50 ±2. 66） nmol ·mg^-1 markedly （P 〈0. 01 and P 〈0. 05）. Furthermore, the activities of SOD （89.55 ±6.93） U ·mg^-1, GSH-Px （845.87 ±63.76） mU ·mg^-1 , CAT （93.07 ±10.40） U ·mg^-1 mg and the content of GSH （ 8.91 ± 1.06 ） μmol·L^-1 of eardiomyocytes were also raised by 600 mg ·L^-1 ASS （ P 〈 0. 05 ）. Conclusion： Taken together, the study implicate that ASS protects cardiomyocytes against oxidative-stress injury induced by H2O2 through reduction of lipid peroxidation and enhancement of the activity of antioxidant defense.