Influence of Sheng-Huang mixture on cardiocyte apoptosis and bax and bcl-2 mRNA expression in a rat model of ischemia-reperfusion injury

doi:10.3969/j.issn.2095-4344.2016.27.014

Abstract

Abstract:

BACKGROUND: Sheng-Huang mixture including Chinese medicine Shengmai Decoction and total flavonoids of stems and leaves of radix has been shown to resist inflammation, regulate immune function, and protect ischemic myocardial tissues. However, its effect on the apoptosis of cardiac muscle cells after ischemia/reperfusion injury remains unclear.
OBJECTIVE: To investigate the effects of Sheng-Huang mixture on cardiocyte apoptosis and bax and bcl-2 mRNA expression in rats with ischemia-reperfusion injury.
METHODS: Thirty-six Sprague-Dawley rats were divided randomly into six groups: low-dose, moderate-dose and high-dose Sheng-Huang mixture, positive control, blank and model groups (n=6). After 7 days of administration, models of myocardial ischemia-reperfusion injury were established. TUNEL was used to detect myocardial apoptosis. RT-PCR was utilized to measure bax and bcl-2 mRNA expression in the ischemic and reperfusion region.
RESULTS AND CONCLUSION: (1) The bcl-2 mRNA expression was significantly higher in the Sheng-Huang mixture group than in the model group (P < 0.05), but bax mRNA expression was significantly lower (P < 0.05). Thus, bcl-2/bax ratio increased. In addition, apoptosis index was more significantly decreased in the Sheng-Huang mixture group (P < 0.05). (2) Results demonstrated that Sheng-Huang mixture can protect rat myocardium against ischemia/reperfusion injury, and effectively increase the bcl-2/bax ratio and inhibit the apoptosis of cardiomyocytes, and the underlying mechanism is mediated by up-regulating bcl-2 mRNA expression and down-regulating bax mRNA expression.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

Key words: Myocardial Ischemia, Apoptosis, Tissue Engineering

CLC Number:

R318

Chen Long, Tian Huan-na, Gao Yu-feng. Influence of Sheng-Huang mixture on cardiocyte apoptosis and bax and bcl-2 mRNA expression in a rat model of ischemia-reperfusion injury[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(27): 4049-4054.

Figures/Tables 1

References

[1] 李健,冯义柏,郎明健.吡格列酮预处理对大鼠心脏缺血再灌注/缺氧再复氧线粒体结构及膜电势的影响[J].中国病理生理杂志,2008, 24(4):720-724.
[2] 邓宇珺,符永恒,谭宁,等.大鼠活体心肌缺血再灌注损伤模型的建立与综合评估[J].中国病理生理杂志,2011,27(2): 412-416.
[3] 薛富善,孙海燕,李平,等.细胞凋亡与心肌缺血再灌注损伤[J].中国医药导刊,2007,9(4):335-339.
[4] Dianat M, Hamzavi GR, Badavi M, et al. Effects of losartan and vanillic Acid co-administration on ischemia-reperfusion-induced oxidative stress in isolated rat heart. Iran Red Crescent Med J. 2014; 16(7):e16664.
[5] Montero MF, Saurim R, Bonservizi WG, et al. Heart injury following intestinal ischemia reperfusion in rats is attenuated by association of ischemic preconditioning and adenosine. Acta Cir Bras. 2014;29 Suppl 2:67-71.
[6] Wang G, Jiang L, Song J, et al. Mipu1 protects H9c2 myogenic cells from hydrogen peroxide-induced apoptosis through inhibition of the expression of the death receptor Fas. Int J Mol Sci. 2014;15(10):18206- 18220.
[7] Saeed SA, Waqar MA, Zubairi AJ, et al. Myocardial ischaemia and reperfusion injury: reactive oxygen species and the role of neutrophil. J Coll Physicians Surg Pak. 2005;15(8):507-514.
[8] Kovacevi? M, Simi? O, Jonji? N, et al. Apoptosis and cardiopulmonary bypass. J Card Surg. 2007;22(2): 129-134.
[9] Zhao ZQ, Corvera JS, Halkos ME, et al. Inhibition of myocardial injury by ischemic postconditioning during reperfusion: comparison with ischemic preconditioning. Am J Physiol Heart Circ Physiol. 2003;285(2):H579- 588.
[10] 郝金凤,郭亚春.生黄合剂对病毒性心肌炎小鼠脾脏Ｔ淋巴细胞影响的研究[J].现代免疫学,2015, 35(4):294-298.
[11] 高玉峰,张凤英,米术斌. 生黄合剂对病毒性心肌炎小鼠的干预实验[J]. 中国临床康复,2006,10(35):51-53.
[12] 周晓春,颜勇,王宝忠,等. 生黄合剂对病毒性心肌炎小鼠血清SOD 活性和MDA 含量的影响[J]. 承德医学院学报,2008,25(1):10-11.
[13] 郭亚春,周晓春,宋鸿儒,等. 黄芩茎叶总黄酮与生脉饮对病毒性心肌炎心脏细胞因子mRNA 表达的影响[J]. 山东医药,2008,48(2):46-48.
[14] 张凤英,高玉峰,宋鸿儒.黄芩茎叶提取物与生脉饮抗柯萨奇病毒B3的体外研究[J]. 天津医药,2005,33(11): 716-718.
[15] Lindsten T, Ross AJ, King A, et al. The combined functions of proapoptotic Bcl-2 family members bak and bax are essential for normal development of multiple tissues. Mol Cell. 2000;6(6):1389-1399.
[16] 刘胜辉,杨新春,范谦.缺血后处理对大鼠缺血再灌注心肌细胞凋亡的影响[J].中国组织工程研究与临床康复, 2007, 11(8):1450-1452.
[17] 陈燕,夏豪,鲍翠玉,等.心肌缺血再灌注损伤模型大鼠心肌组织及血清肌钙蛋白Ⅰ表达和心肌肿瘤坏死因子α与重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白的干预[J]. 中国组织工程研究与临床康复,2011,11(37): 6879-6882.
[18] 解丽君,张建新,李兰芳,等.异丙酚激活蛋白激酶C 对心肌缺血再灌注损伤的保护作用[J]. 中国组织工程研究与临床康复,2007,11(29): 5735-5741
[19] 张慧芝,白建平,于肯明.芹菜素对缺血再灌注大鼠心肌核转录因子-κ B和ICAM-1表达的影响[J].中国组织化学与细胞化学杂志,2010,19(2):181-184.
[20] Xie Z, Koyama T, Suzuki J, et al. Coronary reperfusion following ischemia: different expression of bcl-2 and bax proteins, and cardiomyocyte apoptosis. Jpn Heart J. 2001;42(6):759-770.
[21] 赵永强,隽兆东,管英俊,等.PI3K与ERK1/2信号传导通路在吡格列酮保护大鼠心肌缺血再灌注损伤中的作用机制[J].中国实验诊断学,2012,16(2):207-210.
[22] 贺欣,陶贵周,齐志敏.葡萄糖酸镁对离体大鼠心肌缺血再灌注细胞凋亡的抑制效应[J].中国组织工程研究与临床康复,2007,11(8):1465-1468.
[23] Yang B, Johnson TS, Thomas GL, et al. Expression of apoptosis-related genes and proteins in experimental chronic renal scarring. J Am Soc Nephrol. 2001;12(2): 275-288.
[24] McClintock DS, Santore MT, Lee VY, et al. Bcl-2 family members and functional electron transport chain regulate oxygen deprivation-induced cell death. Mol Cell Biol. 2002;22(1):94-104.
[25] 刘静,牛鹏武.川芎嗪对大鼠心肌缺血再灌注损伤细胞凋亡的影响[J].中国实用医药,2011,6(2):29-30.
[26] 李波,杨萌,刘景伟,等. HRS调控凋亡相关蛋白Bcl-xL、Bad、Bcl-2、Bax对大鼠心肌缺血再灌注损伤影响[J].中国公共卫生,2015,31(7):889-892.
[27] 赵润英,郝伟,孟祥军,等.阿魏酸川芎嗪对缺血再灌注大鼠心肌细胞凋亡及凋亡相关蛋白表达的影响[J].中国病理生理杂志,2013,29(12):2139-2143.
[28] 陈文娟,胡道德,刘亮.中药治疗病毒性心肌炎的实验研究进展[J].中国中西医结合杂志,2013,33(10):1438-1440.
[29] Galagudza M, Kurapeev D, Minasian S, et al. Ischemic postconditioning: brief ischemia during reperfusion converts persistent ventricular fibrillation into regular rhythm. Eur J Cardiothorac Surg. 2004;25(6): 1006-1010.
[30] Gottlieb RA, Burleson KO, Kloner RA, et al. Reperfusion injury induces apoptosis in rabbit cardiomyocytes. J Clin Invest. 1994;94(4):1621-1628.
[31] Kajstura J, Cheng W, Reiss K, et al. Apoptotic and necrotic myocyte cell deaths are independent contributing variables of infarct size in rats. Lab Invest. 1996;74(1):86-107.
[32] 周晓慧,龚明玉,杨鹤梅,等. 黄芩茎叶总黄酮对缺血/复氧诱导心肌细胞凋亡的影响[J].中国中西医结合杂志,2011, 31(6):803-806.
[33] 卜丽梅,关凤英,乔萍,等.没食子酸对大鼠缺血再灌注损伤后细胞凋亡的保护作用及作用机制[J].中国实验诊断学, 2010,14(11):1693-1697.
[34] 闫文翠,王晶,张维娜,等.生黄合剂对大鼠心肌缺血再灌注损伤的干预作用[J].中国老年学杂志,2013,33(23): 5893-5895.
[35] Pchejetski D, Kunduzova O, Dayon A, et al. Oxidative stress-dependent sphingosine kinase-1 inhibition mediates monoamine oxidase A-associated cardiac cell apoptosis Circ Res. 2007;100(1):41-49.
[36] 贺苗,赵杰,苏健,等. 1-磷酸鞘氨醇后适应对大鼠心肌缺血/再灌注损伤的保护作用[J].中国药理学通报,2013, 29(10):1369-1372.