留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

针刺内关和足三里双穴位对大鼠心肌缺血再灌注损伤后的血管重塑作用

张宏如 陈静 徐森磊 顾任钧 白桦 卢圣锋 潘玉璟 顾一煌

张宏如, 陈静, 徐森磊, 顾任钧, 白桦, 卢圣锋, 潘玉璟, 顾一煌. 针刺内关和足三里双穴位对大鼠心肌缺血再灌注损伤后的血管重塑作用[J]. 南京中医药大学学报, 2023, 39(1): 42-49. doi: 10.14148/j.issn.1672-0482.2023.0042
引用本文: 张宏如, 陈静, 徐森磊, 顾任钧, 白桦, 卢圣锋, 潘玉璟, 顾一煌. 针刺内关和足三里双穴位对大鼠心肌缺血再灌注损伤后的血管重塑作用[J]. 南京中医药大学学报, 2023, 39(1): 42-49. doi: 10.14148/j.issn.1672-0482.2023.0042
ZHANG Hong-ru, CHEN Jing, XU Sen-lei, GU Ren-jun, BAI Hua, LU Shen-feng, PAN Yu-jing, GU Yi-huang. The Effect of Combination Acupuncture Treatment of Neiguan and Zusanli on Vascular Remodeling after Myocardial Ischemia Reperfusion Injury[J]. Journal of Nanjing University of traditional Chinese Medicine, 2023, 39(1): 42-49. doi: 10.14148/j.issn.1672-0482.2023.0042
Citation: ZHANG Hong-ru, CHEN Jing, XU Sen-lei, GU Ren-jun, BAI Hua, LU Shen-feng, PAN Yu-jing, GU Yi-huang. The Effect of Combination Acupuncture Treatment of Neiguan and Zusanli on Vascular Remodeling after Myocardial Ischemia Reperfusion Injury[J]. Journal of Nanjing University of traditional Chinese Medicine, 2023, 39(1): 42-49. doi: 10.14148/j.issn.1672-0482.2023.0042

针刺内关和足三里双穴位对大鼠心肌缺血再灌注损伤后的血管重塑作用

doi: 10.14148/j.issn.1672-0482.2023.0042
基金项目: 

国家自然科学基金面上项目 81974583

南京中医药大学校自然青年基金项目 NZY81704169

详细信息
    通讯作者:

    张宏如, 男, 副教授, 主要从事针灸效应机制研究, E-mail: 170947@njucm.edu.cn

  • 中图分类号: R285.5

The Effect of Combination Acupuncture Treatment of Neiguan and Zusanli on Vascular Remodeling after Myocardial Ischemia Reperfusion Injury

  • 摘要:   目的  研究针刺内关和足三里对心肌缺血再灌注损伤后的血管重塑作用。  方法  将SPF级雄性SD大鼠分为空白组(10只)、MIRI组(10只)、内关组(10只)、足三里组(10只)以及内关+足三里组(10只)。对大鼠进行心肌缺血再灌注(MIRI)处理从而构建MIRI动物模型。穴取双侧内关、足三里以及内关+足三里, 疏密波, 频率2 Hz。关胸后第2天进行电针治疗, 每日1次, 共14 d。超声心动图(ECG)检查各组大鼠左心室射血分数(LVEF)及左心室内径缩短率(LVFS)。TTC染色检测各组大鼠心肌梗死面积。免疫荧光法(IF)检测各组大鼠心肌组织中α-SMA和CD31蛋白共表达情况。蛋白免疫印迹法(Western blot)分别检测各组大鼠心肌组织中VEGFA、α-SMA, CD31和p-eNOS蛋白表达水平变化。  结果  与空白组比较, MIRI模型组大鼠LVEF和LVFS参数显著降低(P < 0.000 1)。针刺内关或足三里穴位的大鼠LVEF和LVFS参数较未进行针刺的MIRI大鼠显著升高(P < 0.000 1)。针刺内关和足三里双穴位的大鼠LVEF和LVFS参数相比MIRI模型组(P < 0.000 1)以及针刺单穴位组(P < 0.01)均有显著升高。MIRI组的梗死面积显著高于空白组(P < 0.000 1)。单独针刺内关穴(P < 0.001)和足三里穴(P < 0.01)能显著缩小梗死面积, 而针刺内关穴和足三里双穴位相比针刺单个穴位能更有效缩小梗死面积(P < 0.05, P < 0.001)。与空白组比较, MIRI组大鼠心肌组织中α-SMA和CD31表达阳性显著减弱。相较MIRI组, 单独针刺内关或足三里穴位组的大鼠心肌组织中α-SMA和CD31表达阳性增强, 而针刺内关和足三里双穴位组的表达阳性强于MIRI对照组以及针刺单穴位组。MIRI组的α-SMA、CD31、VEGFA和p-eNOS蛋白表达水平相对空白组显著降低(P < 0.000 1)。单独针刺内关或足三里穴位组的大鼠心肌组织中VEGFA(P < 0.001, P < 0.000 1)、α-SMA(P < 0.000 1)、CD31(P < 0.000 1)和p-eNOS(P < 0.000 1)表达增强, 而针刺内关和足三里双穴位组表达升高更为显著(P < 0.000 1)。  结论  针刺内关和足三里双穴位对心肌缺血再灌注损伤后心血管重塑效果更佳。

     

  • 图  1  超声心动图分析模前后大鼠的心电情况

    Figure  1.  Detect the electrocardiogram of rats in each group by ECG

    图  2  超声心动图检测各组大鼠的LVEF及LVFS (x±s, n=10)

    注: **P < 0.01, ****P < 0.000 1。

    Figure  2.  Detect the LVEF and LVFS by ECG (x±s, n=10)

    图  3  TTC染色检测各组大鼠心肌梗死面积(x±s, n=10)

    注: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1。

    Figure  3.  Detect the infarct area in each groups using TTC staining (x±s, n=10)

    图  4  免疫荧光法检测各组大鼠心肌组织中CD31和α-SMA共表达情况

    Figure  4.  Test the co-expression of α-SMA and CD31 protein in each groups by IF

    图  5  Western blot法检测各组大鼠心肌组织中VEGFA、CD31、α-SMA和p-eNOS蛋白表达变化

    注: ***P < 0.001, ****P < 0.000 1。

    Figure  5.  Measure the protein levels of VEGFA, α-SMA and CD31 in each groups by Western blot

  • [1] ZALEWSKI J, NOWAK K, FURCZYNSKA P, et al. Complicating acute myocardial infarction. current status and unresolved targets for subsequent research[J]. J Clin Med, 2021, 10(24): 5904. doi: 10.3390/jcm10245904
    [2] BOATENG S, SANBORN T. Acute myocardial infarction[J]. Dis Mon, 2013, 59(3): 83-96. doi: 10.1016/j.disamonth.2012.12.004
    [3] LU L, LIU M, SUN RR, et al. Myocardial infarction: Symptoms and treatments[J]. Cell Biochem Biophys, 2015, 72(3): 865-867. doi: 10.1007/s12013-015-0553-4
    [4] SCHUMACHER B, PECHER P, VON SPECHT BU, et al. Induction of neoangiogenesis in ischemic myocardium by human growth factors: First clinical results of a new treatment of coronary heart disease[J]. Circulation, 1998, 97(7): 645-650. doi: 10.1161/01.CIR.97.7.645
    [5] SPATH NB, MILLS NL, CRUDEN NL. Novel cardioprotective and regenerative therapies in acute myocardial infarction: A review of recent and ongoing clinical trials[J]. Future Cardiol, 2016, 12(6): 655-672. doi: 10.2217/fca-2016-0044
    [6] 王贞, 张志国, 张君. 益气通脉解毒方治疗心肌梗死疗效观察及对血管新生指标的影响[J]. 新中医, 2021, 53(8): 47-50. https://www.cnki.com.cn/Article/CJFDTOTAL-REND202108013.htm

    WANG Z, ZHANG ZG, ZHANG J. Curative effect of Yiqi Tongmai Jiedu prescription on myocardial infarction and its effect on angiogenesis indexes[J]. J New Chin Med, 2021, 53(8): 47-50. https://www.cnki.com.cn/Article/CJFDTOTAL-REND202108013.htm
    [7] 刘甜甜, 张彦丽, 何婷, 等. 中药改善心肌梗死促血管新生机制的研究进展[J]. 环球中医药, 2022, 15(8): 1503-1509. doi: 10.3969/j.issn.1674-1749.2022.08.044

    LIU TT, ZHANG YL, HE T, et al. Research progress on the mechanism of traditional Chinese medicine promoting angiogenesis in myocardial infarction[J]. Glob Tradit Chin Med, 2022, 15(8): 1503-1509. doi: 10.3969/j.issn.1674-1749.2022.08.044
    [8] 曾玉玲, 李世阁, 占德进. 参麦注射液促进心肌梗死模型大鼠心肌血管新生[J]. 基础医学与临床, 2021, 41(11): 1624-1628. doi: 10.16352/j.issn.1001-6325.2021.11.011

    ZENG YL, LI SG, ZHAN DJ. Promoting effect of Shenmai injection on myocardial angiogenesis in rat models with myocardial infarction[J]. Basic Clin Med, 2021, 41(11): 1624-1628. doi: 10.16352/j.issn.1001-6325.2021.11.011
    [9] YANG MX, SUN MS, DU T, et al. The efficacy of acupuncture for stable angina pectoris: A systematic review and meta-analysis[J]. Eur J Prev Cardiol, 2021, 28: 1415-1425. doi: 10.1177/2047487319876761
    [10] 覃佩兰. 针刺"内关穴"对心肌缺血小鼠心肌组织氯离子通道CLC-3、CLC-4蛋白表达的影响[D]: 沈阳: 辽宁中医药大学, 2016.

    TAN PL. Effect of acupuncture at Neiguan Point on protein expression of chloride ion channels CLC-3 and CLC-4 in myocardial ischemia mice[D]. Shenyang: Liaoning University of Chinese Medicine, 2016.
    [11] ZHAO L, LI DH, ZHENG H, et al. Acupuncture as adjunctive therapy for chronic stable angina: A randomized clinical trial[J]. JAMA Intern Med, 2019, 179(10): 1388-1397. doi: 10.1001/jamainternmed.2019.2407
    [12] 王华, 卢继东, 吴松, 等. 电针不同腧穴对心肌缺血模型大鼠细胞凋亡及心肌细胞miRNAs表达的影响[J]. 中国针灸, 2016, 36(3): 281-286. doi: 10.13703/j.0255-2930.2016.03.016

    WANG H, LU JD, WU S, et al. Effects of electroacupuncture at different acupoints on apoptosis and the expression of miRNAs in myocardial cells in rats model of myocardial ischemia[J]. Chin Acupunct Moxibust, 2016, 36(3): 281-286. doi: 10.13703/j.0255-2930.2016.03.016
    [13] 付梦婷, 王华, 吴松. 标本配穴电针预处理对心肌缺血再灌注损伤大鼠CTGF表达的影响[J]. 湖北中医药大学学报, 2019, 21(6): 5-8. https://www.cnki.com.cn/Article/CJFDTOTAL-HZXX201906001.htm

    FU MT, WANG H, WU S. Effect of the pretreatment with electric acupuncture of "combined Biao and Ben acupoints" on CTGF in myocardial ischemia-reperfusion injury[J]. J Hubei Univ Chin Med, 2019, 21(6): 5-8. https://www.cnki.com.cn/Article/CJFDTOTAL-HZXX201906001.htm
    [14] 张桂诚, 余剑波. 电针足三里穴和内关穴对兔内毒素休克诱发心肌损伤的影响及血红素氧合酶-1在其中的作用[J]. 中华麻醉学杂志, 2014, 34(2): 211-215.

    ZHANG GC, YU JB. Effect of electro-acupuncture at Zusanli and Neiguan acupoints on endotoxic shock-induced myocardial injury in rabbits and the role of heme oxygenase-1[J]. Chin J Anesthesiol, 2014, 34(2): 211-215.
    [15] COCHAIN C, CHANNON KM, SILVESTRE JS. Angiogenesis in the infarcted myocardium[J]. Antioxid Redox Signal, 2013, 18(9): 1100-1113. doi: 10.1089/ars.2012.4849
    [16] ROBICH MP, CHU LM, OYAMADA S, et al. Myocardial therapeutic angiogenesis: A review of the state of development and future obstacles[J]. Expert Rev Cardiovasc Ther, 2011, 9(11): 1469-1479. doi: 10.1586/erc.11.148
    [17] LOSORDO DW, VALE PR, SYMES JF, et al. Gene therapy for myocardial angiogenesis: Initial clinical results with direct myocardial injection of phVEGF165 as sole therapy for myocardial ischemia[J]. Circulation, 1998, 98(25): 2800-2804. doi: 10.1161/01.CIR.98.25.2800
    [18] YUAN LB, DAI XM, FU HY, et al. Vaspin protects rats against myocardial ischemia/reperfusion injury (MIRI) through the TLR4/NF-κB signaling pathway[J]. Eur J Pharmacol, 2018, 835: 132-139. doi: 10.1016/j.ejphar.2018.07.052
    [19] HU JJ, CHEN XL, LI P, et al. Exosomes derived from human amniotic fluid mesenchymal stem cells alleviate cardiac fibrosis via enhancing angiogenesis in vivo and in vitro[J]. Cardiovasc Diagn Ther, 2021, 11(2): 348-361. doi: 10.21037/cdt-20-1032
    [20] 葛瑜. 温和灸预处理对幼鼠心肌缺血再灌注损伤的影响[J]. 上海针灸杂志, 2021, 40(3): 352-357. https://www.cnki.com.cn/Article/CJFDTOTAL-SHZJ202103021.htm

    GE Y. Effect of pretreatment with mild moxibustion on myocardial ischemia-reperfusion injury in young rats[J]. Shanghai J Acupunct Moxibust, 2021, 40(3): 352-357. https://www.cnki.com.cn/Article/CJFDTOTAL-SHZJ202103021.htm
    [21] 郭颖, 孙兴华, 苏苏, 等. 针刺内关穴对慢性心衰小鼠心脏血流动力学及心衰标志物的影响[J]. 中国中医急症, 2017, 26(3): 387-389. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYJZ201703004.htm

    GUO Y, SUN XH, SU S, et al. Study on the effect of acupuncture on the point of Neiguan on heart hemodynamics and heart failure markers of chronic heart failure rats[J]. J Emerg Tradit Chin Med, 2017, 26(3): 387-389. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYJZ201703004.htm
    [22] 刘柏, 王瑞元, 熊开宇, 等. 针刺内关穴对改善运动中心肌缺血和心脏泵血功能的研究[J]. 体育科学, 1993, 13(6): 61-65, 95. https://www.cnki.com.cn/Article/CJFDTOTAL-TYKX199306020.htm

    LIU B, WANG RY, XIONG KY, et al. A study of the effect of acupuncture of Neiguan point on improving myocardial iachemia and the blood pumping function of the hart during exercise[J]. China Sport Sci, 1993, 13(6): 61-65, 95. https://www.cnki.com.cn/Article/CJFDTOTAL-TYKX199306020.htm
    [23] 唐关敏, 翟昌林, 宋国杰, 等. 针灸联合骨髓间充质干细胞移植对大鼠心肌梗死区心肌重构的影响[J]. 中华危重症医学杂志(电子版), 2014, 7(6): 395-399. https://www.cnki.com.cn/Article/CJFDTOTAL-ZWZD201406007.htm

    TANG GM, ZHAI CL, SONG GJ, et al. Effects of acupuncture combined with bone marrow mesenchymal stem cells on myocardial remodeling in rats after myocardial infarction[J]. Chin J Crit Care Med Electron Ed, 2014, 7(6): 395-399. https://www.cnki.com.cn/Article/CJFDTOTAL-ZWZD201406007.htm
    [24] 孙忠人, 姜松林, 赵瑛. 针刺"足三里"穴对大鼠心脏功能影响的实验研究[J]. 中医药信息, 1988, 5(1): 40-41. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYXN198801026.htm

    SUN ZR, JIANG SL, ZHAO Y. Experimental study on the effect of acupuncture at Zusanli point on cardiac function in rats[J]. Inf Tradit Chin Med, 1988, 5(1): 40-41. https://www.cnki.com.cn/Article/CJFDTOTAL-ZYXN198801026.htm
    [25] 许涛, 曹好好, 许美霞, 等. 电针刺足三里穴对脓毒症大鼠心脏损害机制的研究[J]. 中华实验外科杂志, 2013, 30(8): 1564-1567.

    XU T, CAO HH, XU MX, et al. Effect of electro-acupuncture at Zusanli point on the injury of the heart function in sepsis rats[J]. Chin J Exp Surg, 2013, 30(8): 1564-1567.
    [26] 徐添添, 周建庆. 动脉粥样硬化斑块内血管新生的研究进展[J]. 浙江医学, 2019, 41(4): 393-396. https://www.cnki.com.cn/Article/CJFDTOTAL-ZJYE201904031.htm

    XU TT, ZHOU JQ. Research progress of angiogenesis in atherosclerotic plaque[J]. Zhejiang Med J, 2019, 41(4): 393-396. https://www.cnki.com.cn/Article/CJFDTOTAL-ZJYE201904031.htm
    [27] VAN DER LAAN AM, PIEK JJ, VAN ROYEN N. Targeting angiogenesis to restore the microcirculation after reperfused MI[J]. Nat Rev Cardiol, 2009, 6(8): 515-523. http://www.researchgate.net/publication/26294339_Laan_AM_Targeting_angiogenesis_to_restore_the_microcirculation_after_reperfused_MI_Nat_Rev_Cardiol_68_515-523
    [28] KOBAYASHI K, MAEDA K, TAKEFUJI M, et al. Dynamics of angiogenesis in ischemic areas of the infarcted heart[J]. Sci Rep, 2017, 7(1): 7156.
    [29] LOSORDO DW, DIMMELER S. Therapeutic angiogenesis and vasculogenesis for ischemic disease. part I: Angiogenic cytokines[J]. Circulation, 2004, 109(21): 2487-2491.
    [30] TONGERS J, LOSORDO DW, LANDMESSER U. Stem and progenitor cell-based therapy in ischaemic heart disease: Promise, uncertainties, and challenges[J]. Eur Heart J, 2011, 32(10): 1197-1206.
    [31] 陈春兰, 张运君, 许和平, 等. 藁本内酯通过降低VEGF的水平抑制人血管瘤内皮细胞的血管新生和上皮-间充质转化[J]. 中国病理生理杂志, 2020, 36(8): 1422-1427. https://www.cnki.com.cn/Article/CJFDTOTAL-ZBLS202008012.htm

    CHEN CL, ZHANG YJ, XU HP, et al. Ligustilide inhibits angiogenesis and epithelial-mesenchymal transition in human hemangioendothelial cells by reducing VEGF levels[J]. Chin J Pathophysiol, 2020, 36(8): 1422-1427. https://www.cnki.com.cn/Article/CJFDTOTAL-ZBLS202008012.htm
    [32] 冯燕兵, 熊烈, 石彦波, 等. 眼络通方对兔视网膜静脉阻塞模型促血管新生因子PDGF、VEGF表达的影响[J]. 浙江中医药大学学报, 2020, 44(7): 605-612. https://www.cnki.com.cn/Article/CJFDTOTAL-BHON202007004.htm

    FENG YB, XIONG L, SHI YB, et al. Effect of yanluotong decoction on the expression of proangiogenic factors PDGF and VEGF in rabbits model with retinal vein occlusion[J]. J Zhejiang Chin Med Univ, 2020, 44(7): 605-612. https://www.cnki.com.cn/Article/CJFDTOTAL-BHON202007004.htm
    [33] 胡扬涛. 实验性脉络膜新生血管COX-2、CD31的表达变化及塞来昔布对其影响的研究[D]. 昆明: 昆明医科大学, 2012.

    HU YT. Experimental choroid neovascularization COX-2, changes in the expression of CD31 and celecoxib the study of the impact[D]. Kunming: Kunming Medical College, 2012.
    [34] 魏彦珍. 基于调节α-SMA、CD133的脑脉通对I/R老龄大鼠脑微血管生成的实验与临床研究[D]. 郑州: 河南中医学院, 2015.

    WEI YZ. Based on regulating α-SMA, CD133 cerebral vein access to I/R aging rat microvascular generation experiment and clinical research[D]. Zhengzhou: Henan Medical College, 2015.
    [35] 王医术, 李玉林, 王心蕊, 等. 用α-SMA结合部位标记新生血管周细胞[J]. 吉林大学学报(医学版), 2003, 29(5): 594-595, 598. https://www.cnki.com.cn/Article/CJFDTOTAL-BQEB200305026.htm

    WANG YS, LI YL, WANG XR, et al. Study on α-SMA as a new marker of pericytes combined with location[J]. J Jilin Univ Med Ed, 2003, 29(5): 594-595, 598. https://www.cnki.com.cn/Article/CJFDTOTAL-BQEB200305026.htm
    [36] ZHENG Y, WU Z, YI F, et al. By activating Akt/eNOS bilobalide B inhibits autophagy and promotes angiogenesis following focal cerebral ischemia reperfusion[J]. Cell Physiol Biochem, 2018, 47(2): 604-616.
    [37] HUANG CC, KUO HM, WU PC, et al. Soluble delta-like 1 homolog (DLK1) stimulates angiogenesis through Notch1/Akt/eNOS signaling in endothelial cells[J]. Angiogenesis, 2018, 21(2): 299-312.
  • 加载中
图(5)
计量
  • 文章访问数:  307
  • HTML全文浏览量:  30
  • PDF下载量:  19
  • 被引次数: 0
出版历程
  • 收稿日期:  2022-11-20
  • 网络出版日期:  2023-01-18
  • 发布日期:  2023-01-10

目录

    /

    返回文章
    返回