益气通脉颗粒对高脂大鼠心肌缺血再灌注损伤后心肌细胞凋亡的影响
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摘要
目的:研究益气通脉颗粒对高脂大鼠心肌缺血再灌注损伤后心肌细胞凋亡的影响,并探讨其机制。方法:采用高脂膳食喂养的方法复制高脂大鼠模型,再通过夹闭冠状动脉的方法复制心肌缺血再灌注大鼠模型,选取40只健康成年雄性SD大鼠随机分成假手术组、高脂大鼠缺血再灌注模型组(GI/R)、益气通脉颗粒高、中、低剂量药物治疗组(1.352、0.676、0.338 g/mL)。通过可逆性左冠状动脉前降支结扎法建立心肌缺血再灌注模型,假手术组仅穿线不结扎。益气通脉颗粒高、中、低剂量组大鼠给予不同浓度的益脉颗粒灌胃,3次/d,疗程1周,假手术组和GI/R组给予等量的0.9%氯化钠注射液灌胃。治疗完成后,取腹主动脉血,使用全自动生化分析仪检测血清肌酸激酶同功酶(CK-MB)、乳酸脱氢酶(LDH)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和甘油三酯(TG)的含量;通过TTC染色测定各组大鼠心肌梗死面积;免疫印迹(Western Blot)检测Bcl-2、Bax基因蛋白的表达。结果:与GI/R组比较,益气通脉颗粒各组大鼠心肌组织梗死面积减少,其中高剂量组明显降低(P<0.01);与假手术组相比,GI/R组和益气通脉颗粒各组的血清CK-MB、LDH、TC、LDL-C、TG水平明显升高(P<0.05),与GI/R组比较益气通脉颗粒高剂量组血清CK-MB、LDH、TC、LDL-C、TG水平明显降低(P<0.01或P<0.05);与假手术组大鼠比较,GI/R组大鼠基因蛋白Bax表达增强,基因蛋白Bcl-2表达明显减弱(P<0.01)。与GI/R组大鼠比较,益气通脉颗粒各组大鼠基因蛋白Bax的表达明显下降(P<0.05),基因蛋白Bcl-2表达上调,其中高剂量组表达明显上调(P<0.05)。结论:益气通脉颗粒组能降低缺血再灌注损伤的心肌细胞发生凋亡,其作用机制可能与益气通脉颗粒能够下调基因蛋白Bax表达,上调基因蛋白Bcl-2表达,在心肌缺血再灌注损伤时起到保护心肌细胞的作用。
Objectine:To investigate the effects of Yiqi Tongmai Granule on myocardial apoptosis after myocardial ischemia-reperfusion injury in hyperlipidemia ratsand explore its mechanism. Methods: A high-fat diet was used to replicate the hyperlipidemia rat model. The rat model of myocardial ischemia-reperfusion was replicated by clamping the coronary artery. Forty healthy adult male SD rats were randomly divided into sham operation group,lipid-induced rat ischemia-reperfusion model group(GI/R),Yiqi Tongmai Granules high,middle and low dose groups(1.352、0.676、0.338 g/mL). The myocardial ischemia-reperfusion model was established by reversible left anterior descending coronary artery ligation. The sham operation group only threaded without ligation. Rats in the high, middle and low dose groups of Yiqi Tongmai Granules were given different concentrations of Yimai Granules 3 times a day for 1 week. The sham operation group and GI/R group were given the same amount of 0.9% sodium chloride. The injection was administered by stomach. After the treatment was completed, abdominal aortic blood was taken out, and serum creatine kinase isoenzyme(CK-MB), lactate dehydrogenase(LDH), total cholesterol(TC)and low-density lipoprotein cholesterol and the contents of LDL-C and triglyceride(TG) were detected using an automatic biochemical analyzer. The myocardial infarct size of each group was determined by TTC staining. The expression of Bcl-2 and Bax protein was detected by Western Blot. Results: Compared with the GI/R group, the myocardial infarct size of the rats in Yiqi Tongmai Granules groups decreased, especially in the high-dose group(P<0.01). Compared with the shamoperation group, the GI/R group and Yiqi Tongmai Granules groups' levels of serum CK-MB, LDH, TC, LDL-C and TG were significantly increased(P<0.05). Compared with GI/R group, Yiqi Tongmai Granules high-dose group's serum levels of CK-MB, LDH, TC, LDL-Cand TG were significantly decreased(P<0.01 or P<0.05). Compared with the sham-operated rats, the expression of the gene protein Bax was enhanced in the GI/R group. The expression of protein Bcl-2 was significantly attenuated(P<0.01). Compared with the GI/R group, the expression of the gene protein Bax in Yiqi Tongmai Granules groups was significantly decreased(P<0.05), and the expression of the gene protein Bcl-2 was up-regulated, and the expression in the high-dose group was significantly up-regulated(P<0.05). Conclusion: Yiqi Tongmai Granules can reduce the apoptosis of cardiomyocytes induced by ischemia-reperfusion injury. The mechanism may be that Yiqi Tongmai Granules can down-regulate the expression of gene protein Bax and up-regulate the expression of gene protein Bcl-2. It plays a role in protecting cardiomyocytes during blood reperfusion injury.
Objectine:To investigate the effects of Yiqi Tongmai Granule on myocardial apoptosis after myocardial ischemia-reperfusion injury in hyperlipidemia ratsand explore its mechanism. Methods: A high-fat diet was used to replicate the hyperlipidemia rat model. The rat model of myocardial ischemia-reperfusion was replicated by clamping the coronary artery. Forty healthy adult male SD rats were randomly divided into sham operation group,lipid-induced rat ischemia-reperfusion model group(GI/R),Yiqi Tongmai Granules high,middle and low dose groups(1.352、0.676、0.338 g/mL). The myocardial ischemia-reperfusion model was established by reversible left anterior descending coronary artery ligation. The sham operation group only threaded without ligation. Rats in the high, middle and low dose groups of Yiqi Tongmai Granules were given different concentrations of Yimai Granules 3 times a day for 1 week. The sham operation group and GI/R group were given the same amount of 0.9% sodium chloride. The injection was administered by stomach. After the treatment was completed, abdominal aortic blood was taken out, and serum creatine kinase isoenzyme(CK-MB), lactate dehydrogenase(LDH), total cholesterol(TC)and low-density lipoprotein cholesterol and the contents of LDL-C and triglyceride(TG) were detected using an automatic biochemical analyzer. The myocardial infarct size of each group was determined by TTC staining. The expression of Bcl-2 and Bax protein was detected by Western Blot. Results: Compared with the GI/R group, the myocardial infarct size of the rats in Yiqi Tongmai Granules groups decreased, especially in the high-dose group(P<0.01). Compared with the shamoperation group, the GI/R group and Yiqi Tongmai Granules groups' levels of serum CK-MB, LDH, TC, LDL-C and TG were significantly increased(P<0.05). Compared with GI/R group, Yiqi Tongmai Granules high-dose group's serum levels of CK-MB, LDH, TC, LDL-Cand TG were significantly decreased(P<0.01 or P<0.05). Compared with the sham-operated rats, the expression of the gene protein Bax was enhanced in the GI/R group. The expression of protein Bcl-2 was significantly attenuated(P<0.01). Compared with the GI/R group, the expression of the gene protein Bax in Yiqi Tongmai Granules groups was significantly decreased(P<0.05), and the expression of the gene protein Bcl-2 was up-regulated, and the expression in the high-dose group was significantly up-regulated(P<0.05). Conclusion: Yiqi Tongmai Granules can reduce the apoptosis of cardiomyocytes induced by ischemia-reperfusion injury. The mechanism may be that Yiqi Tongmai Granules can down-regulate the expression of gene protein Bax and up-regulate the expression of gene protein Bcl-2. It plays a role in protecting cardiomyocytes during blood reperfusion injury.
引文
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[5] 马春香,孙媛,陈晓斌,等.丹参酮ⅡA预处理对心肌缺血再灌注损伤保护作用的研究[J].陕西医学杂志,2017,46(10):1358-1359.
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[12] 陈丝,贾连群,宋囡,等.香砂六君子汤对脾虚高脂血症大鼠 dyHDL的影响[J/OL].中国实验方剂学杂志.https://doi.org/10.13422/j.cnki.syfjx.20190737.
[13] 黄永艳.化痰活血方对高脂血症大鼠调脂作用的机制研究[D].武汉:湖北中医学院,2006.
[14] WU LL,ZHANG YY.Cardiovascular Pathphysiology[M].Beijing:Peking university medical press,2009:131-134.
[15] 陈卓,李文杰,杨莺.双参通脉颗粒对大鼠心肌缺血再灌注损伤的保护作用[J].中国中医基础医学杂志,2018,24(10):1384-1388.
[16] 贾连群,杨关林,张哲,等.从“脾主运化”理论探讨膏脂转输与胆固醇逆向转运[J].中医杂志,2013,54(20):1793-1795.
[17] 马丽雅,张辉锋.一氧化氮合酶(NOS)与心血管疾病[J].医学临床研究,2010,27(2):349-352.
[18] 张彦清,刘保江,田首元.丙泊酚对大鼠离体缺血/再灌注心肌细胞凋亡和Bcl-2/Bax蛋白表达的影响[J].中西医结合心脑血管病杂志,2011,9(1):55-57.
[19] Pan Y,Qian JX,Lu SQ,et al.Protective effects of tanshinone IIA sodium sulfonate on ischemia-reperfusion-induced myocardial injury in rats[J].Iran J Basic Med Sci,2017,20(3):308-315.
[20] Hu R,Chen ZF,Yan J,et al.Endoplasmic Reticulum Stress of Neutrophils Is Required for Ischemia/Reperfusion-Induced Acute Lung Injury[J].Journal of Immunology,2015,195(10):4802-4809.
[21] 黄欣玮,赵桂峰,毛静远.中药复方防治心肌缺血再灌注损伤机制的研究进展[J].中西医结合心脑血管病杂志,2016,14(23):2773-2776.
[22] Danial NN.BCL-2 family proteins:critical checkpoints of apoptotic cell death[J].Clin Cancer Res,2007,13(24):7254.
[23] Tsujimoto Y.Cell death regulation by the Bcl-2 protein family in the mitoehondria[J].J Cell Physiol,2003,195(2):158.
[2] Vila-Petroff M,Salas MA,Said M,et al.Ca MKⅡ inhi-bition protects against necrosis and apoptosis in irreversible ischemia-reper-fusion injury[J].Cardiovasc Res,2007,73(4):689-698.
[3] Li S J,Wu YN,Kang Y,et al.Effects of limb atraumatic ischemic preconditioning on apoptosis induced by myocar-dial ischemia-reperfusion injury in rats[J].Chin Pharma-col Bull,2009,25(1):100-104.
[4] 吴以岭.脉络学说构建及其指导血管病变防治研究[J].中国中西医结合杂志,2017,37(2):147-148.
[5] 马春香,孙媛,陈晓斌,等.丹参酮ⅡA预处理对心肌缺血再灌注损伤保护作用的研究[J].陕西医学杂志,2017,46(10):1358-1359.
[6] Liao Z,Liu D,Tang L,et al.Long-term oral resveratrol intake provides nutritional preconditioning against myocardial ischemia/reperfusion injury:involvement of VDAC1 downregulation[J].Mol Nutr Food Res,2015,59(3):454-464.
[7] 王俊东,崔勇,王建国,等.人参皂苷Rh3预处理对大鼠心肌缺血再灌注损伤大鼠心肌组织中caspase-3的作用[J].长春中医药大学学报,2017,33(1):13-15.
[8] Cong XQ,Li Y,Lu N,et al.Resveratrol attenuates the inflammatory reaction induced by ischemia/reperfusion in the rat heart[J].Molecular Medicine Reports,2014,9(6):2528-2532.
[9] 张韵致,王瑞雪,周智.复方丹参片-维生素C配伍保护大鼠心肌缺血再灌注损伤[J].中国生物化学与分子生物学报,2018,34(11):1221-1226.
[10] Schwager J,Richard N,Widmer F,et al.Resveratrol distinctively modulates the inflammatory profiles of immune and endothelial cells[J].BMC Complement Altern Med,2017,17(1):309.
[11] 李维娜,冯玲.高脂血症从脾论治探幽[J].世界中西医结合杂志,2017,12(4):577-580.
[12] 陈丝,贾连群,宋囡,等.香砂六君子汤对脾虚高脂血症大鼠 dyHDL的影响[J/OL].中国实验方剂学杂志.https://doi.org/10.13422/j.cnki.syfjx.20190737.
[13] 黄永艳.化痰活血方对高脂血症大鼠调脂作用的机制研究[D].武汉:湖北中医学院,2006.
[14] WU LL,ZHANG YY.Cardiovascular Pathphysiology[M].Beijing:Peking university medical press,2009:131-134.
[15] 陈卓,李文杰,杨莺.双参通脉颗粒对大鼠心肌缺血再灌注损伤的保护作用[J].中国中医基础医学杂志,2018,24(10):1384-1388.
[16] 贾连群,杨关林,张哲,等.从“脾主运化”理论探讨膏脂转输与胆固醇逆向转运[J].中医杂志,2013,54(20):1793-1795.
[17] 马丽雅,张辉锋.一氧化氮合酶(NOS)与心血管疾病[J].医学临床研究,2010,27(2):349-352.
[18] 张彦清,刘保江,田首元.丙泊酚对大鼠离体缺血/再灌注心肌细胞凋亡和Bcl-2/Bax蛋白表达的影响[J].中西医结合心脑血管病杂志,2011,9(1):55-57.
[19] Pan Y,Qian JX,Lu SQ,et al.Protective effects of tanshinone IIA sodium sulfonate on ischemia-reperfusion-induced myocardial injury in rats[J].Iran J Basic Med Sci,2017,20(3):308-315.
[20] Hu R,Chen ZF,Yan J,et al.Endoplasmic Reticulum Stress of Neutrophils Is Required for Ischemia/Reperfusion-Induced Acute Lung Injury[J].Journal of Immunology,2015,195(10):4802-4809.
[21] 黄欣玮,赵桂峰,毛静远.中药复方防治心肌缺血再灌注损伤机制的研究进展[J].中西医结合心脑血管病杂志,2016,14(23):2773-2776.
[22] Danial NN.BCL-2 family proteins:critical checkpoints of apoptotic cell death[J].Clin Cancer Res,2007,13(24):7254.
[23] Tsujimoto Y.Cell death regulation by the Bcl-2 protein family in the mitoehondria[J].J Cell Physiol,2003,195(2):158.