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Effect of electroacupuncture pretreatment on adenine nucleotides in myocardial tissues of rats with myocardial ischemia-reperfusion injury detected by high performance liquid chromatography (HPLC)

基于HPLC观察电针预处理对心肌缺血再灌注损伤大鼠心肌组织中腺苷酸的影响

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Abstract

Objective: To observe the effect of electroacupuncture (EA) pretreatment on adenine nucleotides in the myocardial tissues of the myocardial ischemia-reperfusion injury (MIRI) rats, and to explore the mechanism of EA pretreatment on myocardial prevention and protection in MIRI rats.

Methods: Forty SPF male Sprague-Dawley (SD) rats were randomly divided into 5 groups: a blank group, a sham operation group, a model group, an EA at Neiguan (PC 6) group and an EA at Hegu (LI 4) group, with 8 rats in each group. Rats in the blank group only received binding to the rat plate, 30 min/time, once a day for 7 d; on the 7th day, rats in the sham operation group were subjected to threading for 40 min at the left anterior descending coronary artery without ligation, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day, rats in the model group were subjected to threading at the left anterior descending coronary artery with ligation, for 40 min before the blood flow was restored, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day of pretreatment with EA at Neiguan (PC 6) or Hegu (LI 4) for 30 min per day (once a day for 7 d), rats in the EA at Neiguan (PC 6) group and EA at Hegu (LI 4) group were subjected to modeling and sample collection same as in the model group. The left ventricular myocardium of the lower left anterior descending coronary artery was collected from rats in all 5 groups. Hematoxylin-eosin (HE) staining and transmission electron microscope (TEM) were used to observe the changes in myocardial pathological morphology. The change in the adenine nucleotide level of myocardial tissue was measured by high performance liquid chromatography (HPLC).

Results: The HE staining and ultrastructure showed that the myocardial injury was severer in the model group compared with the sham operation group. Compared with the model group, the myocardial injury in the EA at Neiguan (PC 6) and the EA at Hegu (LI 4) groups was mild or hardly any. The adenine nucleotide levels in the sham operation group and the model group were all decreased compared with the blank group (all P<0.05); compared with the sham operation group, the adenine nucleotide level of the model group was also decreased, but the difference was not statistically significant (P>0.05); compared with the model group, the adenine nucleotide level in the EA at Neiguan (PC 6) group was increased (P<0.05), and the adenine nucleotide level in the EA at Hegu (LI 4) group was significantly increased (P<0.01). The adenine nucleotide level in the EA at Hegu (LI 4) group was higher than that in the EA at Neiguan (PC 6) group, but the difference was not statistically significant (P>0.05). Compared with the EA at Neiguan (PC 6) group, the levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in the EA at Hegu (LI 4) group were significantly increased (all P<0.01).

Conclusion: Both EA at Neiguan (PC 6) and Hegu (LI 4) can alleviate the pathological damage to myocardium in MIRI rats, and increase the adenine nucleotide level in myocardial tissues, and thus protect MIRI rats. EA at Hegu (LI 4) has a better protective effect than Neiguan (PC 6).

摘要

目的:观察电针(EA)预处理对心肌缺血再灌注损伤(MIRI)大鼠心肌组织中腺苷酸的影响, 探讨EA预处理对 MIRI大鼠心肌预防保护的作用机制。方法:将40 只SPF级雄性Sprague-Dawley (SD)大鼠随机分为5 组, 空白组、 假手术组、模型组、EA内关组和EA合谷组, 每组8 只。空白组仅用鼠板束缚大鼠, 每次30 min, 每天1 次,连续7 d; 假手术组于第7 天在左冠状动脉前降支穿线不结扎, 穿线40 min后取线, 静置60 min后取材; 模型组于第7 天在 左冠状动脉前降支结扎, 40 min后恢复血流, 60 min后取材; EA内关组和EA合谷组每天EA内关或合谷30 min, 每天 1 次, 连续7 d, 于第7 天进行造模和取材, 方法同模型组。收集5 组大鼠的左冠状动脉前降支下段左心室心肌组 织。采用苏木精-伊红染色法(HE)及透射电子显微镜(TEM)检测心肌病理形态学的变化, 用高效液相色谱法(HPLC) 测定心肌组织中腺苷酸的含量变化。结果:在心肌HE染色及超微结构方面, 与假手术组比较, 模型组心肌损伤严 重; 与模型组比较, EA内关组心肌损伤较轻微, EA合谷组心肌损伤亦较轻微, 且趋于正常。在腺苷酸含量方面, 与 空白组比较, 假手术组和模型组腺苷酸含量均降低(均P<0.05); 与假手术组比较, 模型组腺苷酸含量亦降低, 但 差异无统计学意义(P>0.05); 与模型组比较, EA内关组腺苷酸含量升高(P<0.05), EA合谷组腺苷酸含量有明显升 高(P<0.01); 与EA内关组比较, EA合谷组腺苷酸含量高于EA内关组, 但差异无统计学意义(P>0.05); 与EA内关组 比较, EA合谷组中三磷酸腺苷(ATP)、二磷酸腺苷(ADP)和一磷酸腺苷(AMP)含量明显升高(均P<0.01)。结论:EA内 关和合谷均能减轻MIRI大鼠心肌的病理损伤, 升高心肌组织中腺苷酸含量, 对MIRI大鼠具有预防保护作用, 且EA 合谷比EA内关的预防保护效果更佳。

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References

  1. Gao JH, Fu WX, Jin ZG, Yu XC. Acupuncture pretreatment protects heart from injury in rats with myocardial ischemia and reperfusion via inhibition of the ß1-adrenoceptor signaling pathway. Life Sci, 2007, 80(16): 1484–1489.

    Article  CAS  Google Scholar 

  2. Yang L, Yang J, Wang Q, Chen M, Lu Z, Chen S, Xiong L. Cardioprotective effects of electroacupuncture pretreatment on patients undergoing heart valve replacement surgery: a randomized controlled trial. Ann Thorac Surg, 2010, 89(3): 781–786.

    Article  Google Scholar 

  3. You HS, Zhu YN, Dong YL, Lü J. Determination of ATP, ADP and AMP in cardiac muscle of rats with heart failure by HPLC. Zhongguo Yaoshi, 2015, 18(5): 722–724.

    CAS  Google Scholar 

  4. Wu YC, Li YH, Xie SN, Li X, Yu S, He SD. High performance liquid chromatographic determination of adenosine phosphates in the mouse brain. Shoudu Yike Daxue Xuebao, 2011, 32(4): 534–537.

    CAS  Google Scholar 

  5. Gao N, Yang Y, Wang SJ, Rong R, Lin JJ, Wang XP. Determination of contents of adenine nucleotides and energy charge in rat liver tissue treated with aconite roots by HPLC. Zhongguo Shiyan Fangjixue Zazhi, 2010, 16(15): 172–175.

    Google Scholar 

  6. Mao KJ, Li FQ, Bai GG, Li X, Chen JW. Simultaneous determination of ten nucleoside and nucleobase components in Hulisan tablets by HPLC. Yaowu Fenxi Zazhi, 2014, 34(11): 1959–1963.

    CAS  Google Scholar 

  7. Wang YQ, Jiang SS, Liu P, Mao GP. Analysis of electrocardiogram changes on myocardial ischemiareperfusion model in rats. Yixue Yanjiusheng Xuebao, 2006, 19(8): 700–702.

    Google Scholar 

  8. Li ZR. Experimental Acupuncture Science. Beijing: China Press of Traditional Chinese Medicine, 2007: 255–257.

    Google Scholar 

  9. Wang C, Zhang JL, Liu Z, Liu M, Yan J, Chang XR, Guo LN, Ai K. Effect of pretreatment with electroacupuncture at point Neiguan (PC 6) on CK and myocardial cell activity in rabbit myocardial ischemia and reperfusion. Shanghai Zhenjiu Zazhi, 2015, 34(1): 67–69.

    Google Scholar 

  10. Tan SJ, Yu Z, Du LD, Chen BC, Dong QT, Wang XQ. HPLC determination of ATP, ADP and AMP in skeletal muscle of rats of postoperative fatigue syndrome. Yaowu Fenxi Zazhi, 2011, 31(11): 2025–2029.

    CAS  Google Scholar 

  11. Xing LL, Guan YX. Reversed-phase high performance liquid chromatography for the detection of ATP and ADP in organisms. Keji Xinxi (Kexue Jiaoyan), 2007, 21(34): 59–60.

    Google Scholar 

  12. Shen W, Vatner DE, Vatner SF, Ingwall JS. Progressive loss of creatine maintains a near normal DeltaG approximately (ATP) in transgenic mouse hearts with cardiomyopathy caused by overexpressing Gsalpha. J Mol Cell Cardiol, 2010, 48(4): 591–599.

    Article  CAS  Google Scholar 

  13. Ebrahimi H, Badalzadeh R, Mohammadi M, Yousefi B. Diosgenin attenuates inflammatory response induced by myocardial reperfusion injury: role of mitochondrial ATP-sensitive potassium channels. J Physiol Biochem, 2014, 70(2): 425–432.

    Article  CAS  Google Scholar 

  14. Li L, Zhang L, Shao Y. Effect on myocardial ischemiareperfusion injury and inflammatory cytokines levels by adenosine preconditioning. Linchuang Mazuixue Zazhi, 2002, 18(1): 11–13.

    CAS  Google Scholar 

  15. Li XJ, Wang X. AMP579 versus adenosine in myocardial ischemia reperfusion injury. Chin Reme Clin, 2015, 15(1): 19–22.

    Google Scholar 

  16. Yan J, Yang XF, Yi SX, Chang XR, Lin YP, Wang C, Diao LH. Effect of electroacupuncture of Neiguan (PC 6) on Na+-K+-ATPase activity and its gene expression in cardiocyte membrane in rats with myocardial ischemiareperfusion injury. Zhen Ci Yan Jiu, 2007, 32(5): 296–300.

    PubMed  Google Scholar 

  17. Wang C, Tian YF, Zhou D, Yang XF, Lin YP, Chang XR, Yan J. Influence of electroacupuncture on myocardial NO and NOS and intracellular Ca2+ contents in myocardial ischemia-reperfusion injury rats. Zhen Ci Yan Jiu, 2010, 35(2): 113–117.

    CAS  PubMed  Google Scholar 

  18. Yang JJ, Yan J, Wang C, Chang XR, Liu M, Lin HB, Xie WJ, He XC, Zhang JL. The effect of acupuncture at Neiguan (PC 6) pretreatment on the expression of blood serum NO, NOS, and adenosine in rabbits with MIR. Zhongguo Zhongyi Jizheng, 2014, 23(7): 1209–1211, 1227.

    Google Scholar 

  19. Tan CF, Yan J, Wang C, Chang XR, Xie WJ, Yang JJ, Liu M, Lin HB, He XC. Effects of electroacupuncture and moxibustion pretreatment on expressions of HSP 27, HSP 70, HSP 90 at different time-points in rabbits with myocardial ischemia-reperfusion injury. Zhen Ci Yan Jiu, 2017, 42(1): 31–38.

    PubMed  Google Scholar 

  20. Tang YN, Tan CF, Liu WW, Yan J, Wang C, Liu M, Lin DH, Huang CH, She L, Chen ML, Li JL, Zhu DM. Effect of electroacupuncture at Neiguan (PC 6) on serum and myocardial metabolites in rats with myocardial ischemia reperfusion injury based on nuclear magnetic resonance spectroscopy. Zhen Ci Yan Jiu, 2018, 43(3): 152–162.

    PubMed  Google Scholar 

  21. Tan CF, Du L, Li JL, Liu WW, Song J, Feng G, Wang C, Yan J, Yang JJ, Tang YN, Chen ML. Effect of electroacupuncture at Neiguan (PC6) on expression of adenosine A1 receptors and content of adenosine in rats with myocardial ischemia-reperfusion injury. Liaoning Zhongyi Zazhi, 2018, 45(3): 641–645.

    Google Scholar 

  22. Tan CF, Wang C, Du L, Liu WW, Song J, Feng G, Yan J, Yang JJ, Tang YN, Chen ML, Li JL. Effect of electroacupuncture and moxibustion pretreatment on expression of autophagy related proteins LC 3 and beclin 1 in rats with myocardial ischemia-reperfusion injury. Zhen Ci Yan Jiu, 2018, 43(1): 1–7.

    CAS  PubMed  Google Scholar 

  23. Szokodi I, Horkay F, Kiss P, Selmeci L, Merkely B. Characterization and stimuli for production of pericardial fluid atrial natriure tic peptide in dogs. Life Sci, 1997, 61(14): 1349–1359.

    Article  CAS  Google Scholar 

  24. Nkere UU, Whawell SA, Thompson EM, Thompson JN, Taylor KM. Changes in pericardial morphology and fibrinolytic activity during cardiopulmonary bypass. J Thorac Cardiovasc Surg, 1993, 106(2): 339–345.

    CAS  PubMed  Google Scholar 

  25. Xie L. Effect of Autologous Pericardium Transplantation on Cardiac Electrical Activity. Chongqing: Master Thesis of Chongqing Medical University, 2014.

    Google Scholar 

  26. Wang XM, Xu LH, Sun GZ, Meng Q. Protective role of pericardial tissue on injured myocardium in rats. Zhonghua Yixue Zazhi, 2011, 91(42): 3012–3015.

    CAS  PubMed  Google Scholar 

  27. Cong YY. Protective Effect and Its Mechanism of High Thoracic Segment Epidural Anesthesia on Myocardium Against Ischemic Reperfusion Injury in Rats. Harbin: Master Thesis of Harbin Medical University: 2008.

    Google Scholar 

Download references

Acknowledgements

This work was supported by National Natural Science Foundation of China (国家自然科学基金项目, No. 81574080); Project of Hunan Province Administration of Traditional Chinese Medicine (湖南省中医药管理局课题, No. 201522); Open Fund Project of Hunan University Innovation Platform (湖南省高校创新平台开放基金项目, No. 15k093).

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  1. Hunan University of Chinese Medicine, Changsha, 410007, China

    Jiao-lan Li  (李姣兰), Chao Wang  (王超), Wei Zhang  (章薇), Cheng-fu Tan  (谭成富), Wei-wei Liu  (刘薇薇), Lin Du  (杜琳), Mei-lin Chen  (陈美琳), Ya-ni Tang  (唐雅妮) & Ding-ming Zhu  (朱鼎铭)

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  1. Jiao-lan Li  (李姣兰)
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Correspondence to Chao Wang  (王超).

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Li, Jl., Wang, C., Zhang, W. et al. Effect of electroacupuncture pretreatment on adenine nucleotides in myocardial tissues of rats with myocardial ischemia-reperfusion injury detected by high performance liquid chromatography (HPLC). J. Acupunct. Tuina. Sci. 17, 289–296 (2019). https://doi.org/10.1007/s11726-019-1127-6

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