Yue R C, Yang X L, Zhang R Y, Liu S, Liu J, Zeng J, Liang H, Wang W, Hu H X, Zeng C Y
Department of Cardiology, North Sichuan Medical College First Affiliated Hospital, Nanchong 637000, China.
Zhonghua Xin Xue Guan Bing Za Zhi. 2017 Jan 25;45(1):57-63. doi: 10.3760/cma.j.issn.0253-3758.2017.01.011.
To investigate the possible mechanism related to the protective effects of salvianolate in H9c2 cells underwent hypoxia/reoxygenation (H/R)-injury. H9c2 cells were divided into four groups: control group, salvianolate group (S group), H/R group, and salvianolate+ H/R group(S+ H/R group), in which the H9c2 cells were pretreated with salvianolate before H/R-treatment.Apoptotic cells were detected by Tunel assays and AnnexinⅤ-FITC apoptosis detection kit.The intracellular ATP level, the change of mitochondrial membrane potential and the mitochondrial DNA oxidative damage were also determined in these groups. (1) The apoptosis rate of H/R group(26.36±5.14)% was significantly higher compared to control group(2.71±1.66)%(=0.000 4), which could be significantly reduced in S+ H/R group(17.28±4.75)%(=0.012 8 vs. H/R group , =0.003 9 vs. control group). The ratio of AnnexinⅤ and PI double positive cells in H/R group(28.23±6.73)% was significantly higher compared to control group(3.53±2.83)%(=0.001 1), which was significantly reduced in S+ H/R group(18.10±4.56)%(=0.037 2 vs. H/R group, =0.038 3 vs. control group). (2)The ATP level of H9c2 cells in H/R group(49.05±10.12)% was significantly lower than in control group 100%(=0.000 5), which was significantly increased in S+ H/R group(68.67±13.32)%(=0.019 9 vs. H/R group). Confocal microscope showed that red fluorescence was dominant in the control group, red fluorescence was significantly reduced, while green fluorescence was significantly increased in H9c2 cells of H/R group and the fluorescence ratio of red to green in H/R group((37.13±8.47)%) was significantly decreased compared to control group (100%, =0.000 1), fluorescence ratio of red to green was significantly increased in S+ H/R group((63.77±12.32)% vs. H/R group, =0.007 3). (3)The mitochondrial DNA oxidative damage in different groups: there was only few 8-hydroxyguanine (8-OHdG) expression, which marked as green, in control group, and 8-OHdG expression was significantly upregulated in H/R group, moreover, the 8-OHdG was co-localized with mitochondria.The expression of 8-OHdG was significantly lower in S+ H/R group compared to H/R group. Salvianolate can reduce mitochondrial DNA oxidative damage, and protect mitochondrial function, thus inhibit myocardial cell apoptosis and eventually reduce the myocardial H/R-injury in H9c2 cells.
为研究丹酚酸对经历缺氧/复氧（H/R）损伤的H9c2细胞的保护作用的可能机制。将H9c2细胞分为四组：对照组、丹酚酸组（S组）、H/R组和丹酚酸 + H/R组（S + H/R组），其中H9c2细胞在H/R处理前用丹酚酸预处理。通过Tunel检测法和AnnexinⅤ-FITC凋亡检测试剂盒检测凋亡细胞。还测定了这些组细胞内ATP水平、线粒体膜电位变化和线粒体DNA氧化损伤情况。（1）H/R组的凋亡率（26.36±5.14）%显著高于对照组（2.71±1.66）%（P = 0.000 4），而S + H/R组（17.28±4.75）%可显著降低凋亡率（与H/R组相比P = 0.012 8，与对照组相比P = 0.003 9）。H/R组AnnexinⅤ和PI双阳性细胞比例（28.23±6.73）%显著高于对照组（3.53±2.83）%（P = 0.001 1），S + H/R组（18.10±4.56）%显著降低（与H/R组相比P = 0.0
