益气养阴通络法对急性心肌梗死大鼠心肌细胞凋亡Bax、Bcl-2、Caspase3表达的影响
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摘要
目的:观察益气养阴通络法对大鼠心肌细胞凋亡Bax、Bcl-2和Caspase3表达的影响,并探讨其作用机制。方法:健康SD大鼠60只,随机分为6组,正常组、模型组、西药组(盐酸地尔硫卓片)、中药低剂量组、中药中剂量组、中药高剂量组。对模型组、中药组、西药组实验动物进行造模,复制急性心肌缺血的模型。西药组及中药组分别给予盐酸地尔硫卓片、中药双参通脉颗粒灌胃2周。采用Western Blotting法检测各组心肌组织中Bax、Bcl-2、Caspase3的表达水平。结果:与正常组比较,模型组Bcl-2表达均显著降低,Bax、Caspase3表达升高(P<0.01或P<0.05)。与模型组比较,各组Bcl-2表达均显著升高,Bax、Caspase3表达均降低(P<0.01或P<0.05)。与模型组比较,中药治疗组中,高剂量组Bcl-2表达明显升高,Bax、Caspase3表达明显降低(P<0.05);西药组与模型组相比,Bcl-2表达明显升高(P<0.01),Caspase3表达明显降低(P<0.05),Bax表达有降低趋势(P>0.05)。中药组高剂量组与西药组相比,Bcl-2表达无明显差异(P>0.05)。结论:益气养阴通络法能够上调急性心肌梗死大鼠心肌细胞Bcl-2表达,下调Bax、Caspase3表达。
Objective:To observe the effect of supplementing Qi,nourishing Yin and removing meridian obstruction method on cardiomyocyte apoptosis and Bax,Bcl-2 and Caspase3 protein expression in rats and explore its mechanism. Methods: Sixty healthy SD rats were randomly divided into 6 groups: normal group, model group, western medicine group(diltiazem hydrochloride tablets), low dose group, medium dose group and high dose group. Experimental animals in the model group, traditional Chinese medicine group and western medicine group were modeled to replicate the model of acute myocardial ischemia. Western medicine group was given diltiazem hydrochloride and the traditional Chinese medicine(TCM)group was given Shuangshen tongmai particals for 2 weeks. The levels of Bax, bcl-2 and Caspase3 in myocardial tissues of each group were determined by Western Blotting.Results: Compared with the normal group,Bc1-2 expression was significantly decreased in each group, and Bax and Caspase3 expressions were increased in the model group(P<0.01 or P<0.05).Compared with the model group, Bc1-2 expression was significantly increased in each group, and Bax and Caspase3 expressions were decreased(P<0.01 or P<0.05).Compared with the model group, Bcl-2 expression was significantly increased in the medium dose group and high dose group, Bax and Caspase3 expressions were decreased in the medium dose group and high dose group(P<0.05). Compared with the model group, the expression of Bc1-2 in the western medicine group was significantly increased(P<0.01).The expression of Caspase3 decreased significantly(P<0.05). Bax expression showed a decreasing trend(P>0.05). There was no significant difference in the expression of Bc1-2 between the highdose Chinese medicine group and the western medicine group(P>0.05). Conclusion: Supplementing Qi,nourishing Yin and removing meridian obstruction method can inhibit the apoptosis of myocardial cells.The mechanism may be related to up-regulate the expression of Bc1-2 and down-regulate the expressions of Bax and Caspase3.
Objective:To observe the effect of supplementing Qi,nourishing Yin and removing meridian obstruction method on cardiomyocyte apoptosis and Bax,Bcl-2 and Caspase3 protein expression in rats and explore its mechanism. Methods: Sixty healthy SD rats were randomly divided into 6 groups: normal group, model group, western medicine group(diltiazem hydrochloride tablets), low dose group, medium dose group and high dose group. Experimental animals in the model group, traditional Chinese medicine group and western medicine group were modeled to replicate the model of acute myocardial ischemia. Western medicine group was given diltiazem hydrochloride and the traditional Chinese medicine(TCM)group was given Shuangshen tongmai particals for 2 weeks. The levels of Bax, bcl-2 and Caspase3 in myocardial tissues of each group were determined by Western Blotting.Results: Compared with the normal group,Bc1-2 expression was significantly decreased in each group, and Bax and Caspase3 expressions were increased in the model group(P<0.01 or P<0.05).Compared with the model group, Bc1-2 expression was significantly increased in each group, and Bax and Caspase3 expressions were decreased(P<0.01 or P<0.05).Compared with the model group, Bcl-2 expression was significantly increased in the medium dose group and high dose group, Bax and Caspase3 expressions were decreased in the medium dose group and high dose group(P<0.05). Compared with the model group, the expression of Bc1-2 in the western medicine group was significantly increased(P<0.01).The expression of Caspase3 decreased significantly(P<0.05). Bax expression showed a decreasing trend(P>0.05). There was no significant difference in the expression of Bc1-2 between the highdose Chinese medicine group and the western medicine group(P>0.05). Conclusion: Supplementing Qi,nourishing Yin and removing meridian obstruction method can inhibit the apoptosis of myocardial cells.The mechanism may be related to up-regulate the expression of Bc1-2 and down-regulate the expressions of Bax and Caspase3.
引文
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[17] 尹智勇,杨俊元,祁宏.Bcl-2蛋白质家族调控细胞凋亡机制的研究进展[J].信阳师范学院学报(自然科学版),2017,30(2):340-344.
[18] Kirshenbaaum LA,Moissac D.The bcl-2 gene product prevents program cell death of ventricular myocytes[J].Circulation,1997,96(7):1580-1585.
[19] 徐少杰,刘松,王宝魁,等.伊伐布雷定对兔急性心梗后心肌细胞凋亡及bcl-2与bax蛋白表达的影响[J].青岛大学医学院学报,2013,49(2):131-133.
[20] Guven BA,Ercan E,Asgun HF,et al.Experimental acute myocardial infarction in rats:HIF-1α,caspase-3,erythropoietin and erythropoietin receptor expression and the cardioprotective effects of two different erythropoietin doses[J].Acta Histochemica,2013,115(7):658-668.
[21] Schwarz K,Simonis G,Yu X,et al.Apoptosis at a distance:remote activation of caspase-3 occurs early after myocardial infarction[J].Molecular and Cellular Biochemistry,2006,281(1):45-54.
[2] 王卫东,陈正堂.Bcl-2/Bax 比率与细胞“命运”[J].中国肿瘤生物治疗杂志,2007,14(4):395.
[3] Brentnall M,Rodriguez-Menocal L,Guevara RLD,et al.Caspase-9,caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis[J].BMC Cell Biology,2013,14(1):32.
[4] 邓铁涛.冠心病的辨证论治[C]//中华中医药学会第十次中医药防治老年病学术交流会论文集,中华中医药学会,2012:2.
[5] 尹丽丽.人参的药理作用及现代研究进展[J].中医临床研究,2003,5(5):122.
[6] 姜文大,梁灵君,顾洁莹,等.人参皂苷Rgl对腹主动脉缩窄致大鼠心肌肥厚和TLR4的影响[J].中药药理与临床,2013,29(1):35-38.
[7] 朱瑄.黄芪的现代药理研究及其临床新用[J].中国中医药现代远程教育,2010,15(8):72.
[8] 代晓光,苏长兰.丹参化学成分及药理研究进展[J].中医药信息,2018,35(4):126-129.
[9] 吴素芬,余日跃,周俊,等.析因设计与中医补气生血方剂最佳药效配伍的研究[J].中国实验方剂学杂志,2011,7(8):153-154.
[10] Meier P,Finch A,Evan G.Apoptosis in development[J].Nature,2000,407(6805):796.
[11] Qiu H,Liu JY,Wei D,et al.Cardiac-generated prostanoids mediate cardiac myocyte apoptosis after myocardial ischaemia[J].Cardiovasc Res,2012,95(3):336-345.
[12] 李敏,王硕仁,赵明镜,等.活血益气方药对心肌梗死后心力衰竭大鼠心肌细胞凋亡的影响[J].中西医结合心脑血管病杂志,2005,3(1):33-35.
[13] Bibbinsdomingo K,Coxson P,Pletcher MJ,et al.Adolescent Overweight and Future Adult Coronary Heart Disease[J].New England Journal of Medicine,2007,357(23):2371-2379.
[14] Hoole SP,Heck PM,Sharples L,et al.Cardiac Remote Ischemic Preconditioning in Coronary Stenting (CRISP Stent) Study A Prospective,Randomized Control Trial [J].Circulation,2009,119(6):820-827.
[15] Ola MS,Nawaz M,Ahsan H.Role of Bcl-2 family proteins and caspases in the regulation of apoptosis[J].Mol Cell Biochem,2011,351:41-58.
[16] Kvansakul M,Hinds MG.The Bcl-2 family:structures,interactions and targets for drug discovery[J].Apoptosis,2015,20(2):136-150.
[17] 尹智勇,杨俊元,祁宏.Bcl-2蛋白质家族调控细胞凋亡机制的研究进展[J].信阳师范学院学报(自然科学版),2017,30(2):340-344.
[18] Kirshenbaaum LA,Moissac D.The bcl-2 gene product prevents program cell death of ventricular myocytes[J].Circulation,1997,96(7):1580-1585.
[19] 徐少杰,刘松,王宝魁,等.伊伐布雷定对兔急性心梗后心肌细胞凋亡及bcl-2与bax蛋白表达的影响[J].青岛大学医学院学报,2013,49(2):131-133.
[20] Guven BA,Ercan E,Asgun HF,et al.Experimental acute myocardial infarction in rats:HIF-1α,caspase-3,erythropoietin and erythropoietin receptor expression and the cardioprotective effects of two different erythropoietin doses[J].Acta Histochemica,2013,115(7):658-668.
[21] Schwarz K,Simonis G,Yu X,et al.Apoptosis at a distance:remote activation of caspase-3 occurs early after myocardial infarction[J].Molecular and Cellular Biochemistry,2006,281(1):45-54.