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

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

    摘要
  • 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。

     

    Abstract:
      OBJECTIVE  Study on the effect of combination treatment of "Neiguan"(PC6) and "Zusanli"(ST36) on vascular remodeling after myocardial Ischemia reperfusion injury (MIRI).
      METHODS  SPF-grade male SD mice were divided into five groups, including Blank (ten rats), MIRI (ten rats), PC6 (ten rats), ST36 (ten rats) and PC6+ST36 (ten rats). All mice were treated with MIRI to establish MIRI animal model. AMI mice were acupunctured bilaterally at PC6 and ST36 respectively or simultaneously, with dilatational wave, 2 Hz in frequency. Two days after suturing, mice were treated with electroacupuncture for 14 days, once a day. Echocardiography (ECG) was used to detect the left ventricular ejection fraction (LVEF) and Left Ventricular Fraction Shortening (LVFS). The infarct area in four groups was measured using TTC staining. Immunofluorescence (IF) was utilized to test the co-expression of α-SMA and CD31 protein in four groups. Western blot (WB) was applied to measure the protein levels of VEGFA, α-SMA and CD31 in five groups.
      RESULTS  Compared to blank group, the levels of LVEF and LVFS in MIRI group was significantly decreased (P < 0.000 1). The levels of LVEF and LVFS were increased in PC6 group and ST36 group (P < 0.000 1) compared to MIRI group. The levels of LVEF and LVFS were highest in PC6+ST36 group (P < 0.000 1 vs MIRI, P < 0.01 vs PC6 or ST36). The infarct area in MIRI group was significantly larger than blank group (P < 0.000 1). The infarct area in PC6 group and ST36 group was significantly smaller than MIRI group (P < 0.001, P < 0.01). The infarct area was reduced into a smallest level in PC6+ST36 group (P < 0.05 vs PC6, P < 0.001 vs ST36). The positivity of α-SMA and CD31 was reduced in MIRI group compared to blank group. Compared to MIRI group, the positivity of α-SMA and CD31 was enhanced in PC6 group and ST36 group, but was highest in PC6+ST36 group. The levels of α-SMA, CD31, VEGFA and p-eNOS in MIRI group were decreased significantly compared to blank group (P < 0.000 1). The levels of VEGFA (P < 0.001, P < 0.000 1 vs MIRI group), α-SMA (P < 0.000 1 vs MIRI group), CD31 (P < 0.000 1 vs MIRI group), p-eNOS (P < 0.000 1 vs MIRI group) were increased when acupunctured at PC6 or ST36, while acupuncture at both PC6 and ST36 had most significant effect on the levels of VEGFA, α-SMA, CD31 and p-eNOS (P < 0.000 1 vs PC6 or ST36).
      CONCLUSION  Acupuncture at both PC6 and ST36 has a better therapeutic effect on vascular remodeling after MIRI.

     

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