Electroacupuncture preconditioning alleviates myocardial ischemia-reperfusion injury through the hypothalamic paraventricular nucleus- interposed nucleus nerve pathway

doi:10.19852/j.cnki.jtcm.2022.03.005

Abstract

Abstract:

OBJECTIVE: To explore whether the paraventricular nucleus (PVN) participates in regulation of the anti-myocardial ischemia-reperfusion injury (MIRI) effect of electroacupuncture (EA) and whether this is achieved through the PVN-interposed nucleus (IN) neural pathway.

METHODS: The modeling method of myocardial ischemia reperfusion injury was achieved by ligating the left anterior descending coronary artery in Sprague-Dawley rats. We used the Powerlab multi-channel physiological recorder system to record electro-cardiograms and analyze the changes in ST segment displacement; 2,3,5-Triphenyltetrazolium chloride staining was used to observe the percentage of myocardial infarction areas. Detecting cardiac troponin I (cTnI), lactate dehydrogenase (LDH) in serum was done with an enzyme-linked immunosorbent assay kit. Morphological changes in the myocardium were detected in each group with hematoxylin-eosin staining of paraffin sections. Detection of c-fos protein expression in the PVN of the hypothalamus was done with the immune-ofluorescence method. The Plexon multi-channel acquisition system recorded PVN neuron discharges and local field potentials in each group of rats. Offline Sorter software was used for cluster analysis. Neuro Explorer software was used to perform autocorrelation, raster and frequency characteristics and spectral energy analysis of neuron signals in each group.

RESULTS: Compared with the MIRI model group, the areas of myocardial infarction in the EA group were significantly reduced; the expression of cTnI, LDH in serum was decreased significantly. The firing frequency of pyramidal cells in the PVN was significantly increased and the spectrum energy map showed energy was reduced, c-fos expression in PVN was reduced, this indicated that neuronal activity in the PVN participates in the effect of EA improving myocardial injury. In addition, we used the kainic acid method to lesion the IN and observed that the effect of EA was weakened. For example, the area of myocardial infarction of lesion IN + EA group in rats was significantly increased compared with that resulting from EA group, the expression of cTnI, LDH in serum was significantly increased, the firing frequency of pyramidal cells in the PVN was significantly reduced. A spectral energy diagram shows that the energy after damage was higher than that of EA group. At the same time, the expression of c-fos in the PVN increased again.

CONCLUSION: Our results indicated that the PVN-IN nerve pathway may participate as an effective pathway of EA to improve the effect of myocardial injury.

Key words: acupuncture, myocardial reperfusion injury, L-lactate dehydrogenase, troponin I, paraventricular hypothalamic nucleus, interposed nucleus

WEI Xiaotong, LI Liaoyuan, ZHANG Yating, SHU Qi, WANG Shuaiya, CHEN Pianpian, HU Ling, YU Qing, CAI Ronglin. Electroacupuncture preconditioning alleviates myocardial ischemia-reperfusion injury through the hypothalamic paraventricular nucleus- interposed nucleus nerve pathway[J]. Journal of Traditional Chinese Medicine, 2022, 42(3): 379-388.

Figures/Tables 9

Figure 1 Changes of ST segment in each group at different time periods A: sham group before ligation; B: sham group received an empty needle; C: sham group was vacuumed for 2 h; D: MIRI group before ligation; E: MIRI group was ligation for 30 min; F: MIRI group was reperfused for 2 h; G: EA group before ligation; H: EA group was ligation for 30 min; I: EA group was reperfused for 2 h; J: lesion +EA group before ligation; K: lesion +EA group was ligation for 30 min L. Lesion + EA group was reperfused for 2 h. The MIRI group refers to the myocardial ischemia reperfusion injury model group. EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group. The lesion + EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture.

Table 1 Statistical analysis of electrocardiogram of rats in each group (`x ± s)

Group	n	Before ligation	Ligation for 30 min	Reperfusion 2 h
Sham	6	0.07±0.05	0.10±0.10	0.07±0.02
MIRI	6	0.10±0.04	0.62±0.17^ad	0.28±0.07^bd
EA	6	0.07±0.03	0.31±0.05^aef	0.14±0.04^cef
Lesion + EA	6	0.10±0.05	0.45±0.13^a	0.21±0.04^c

Figure 2 Statistical analysis of myocardial infarction size, cTnI and LDH between groups A: myocardial infarction area of each group (red is normal myocardial tissue, white is infarct area); B: percentage of myocardial infarction area in each group; C, D: comparison of cTnI and LDH in each group. The MIRI group refers to the myocardial ischemia reperfusion injury model group, EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group, the lesion + EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture; cTnI: cardiac troponin I; LDH: Lactate dehydrogenase. Compared with the sham group, aP < 0.01; compared with the MIRI group, bP < 0.01 and compared with the lesion + EA group, cP < 0.05.

Figure 3 Hematoxylin-eosin staining results in each group A: sham group; B: MIRI group; C: EA group; D: lesion + EA group. The MIRI group refers to the myocardial ischemia reperfusion injury model group, EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group, the lesion + EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture. Partial image is 400 times, scale is 20 μm.

Figure 4 Autocorrelation analysis of PVN neuron electrical activity in each group of rats A: sham group; B: MIRI group; C: EA group; D: lesion + EA group. The MIRI group refers to the myocardial ischemia reperfusion injury model group, EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group, the lesion +EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture.

Figure 5 Comparison of raster images of PVN neuron electrical activity in rats from each group A: sham group; B: MIRI group; C: EA group; D: lesion + EA group. The MIRI group refers to the myocardial ischemia reperfusion injury model group, EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group, the lesion +EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture.

Figure 6 PVN neuron spike counts in each group. A: number of firings of pyramidal cells; B: number of firings of interneurons. The MIRI group refers to the myocardial ischemia reperfusion injury model group, EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group, the lesion + EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture. Compared with the sham group, aP < 0.01; compared with the MIRI group, bP < 0.01; compared with the lesion+ EA group, cP < 0.01.

Figure 7 Comparison of spectral energy diagrams between the groups (time = 300 s; frequency = 40 Hz). A: sham group; B: MIRI group; C: EA group; D: lesion + EA group. The MIRI group refers to the myocardial ischemia reperfusion injury model group, EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group, the lesion + EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture.

Figure 8 c-fos expression in the PVN of each group (immunofluorescence staining, partial image was 200 times, scale was 50 μm) A: sham group; B: MIRI group; C: EA group; D: lesion + EA group. The MIRI group refers to the myocardial ischemia reperfusion injury model group, EA group refers to the electroacupuncture of the heart meridians group + myocardial ischemia-reperfusion injury group, the lesion + EA group refers to lesion of the intercerebellar nucleus + electroacupuncture pretreatment + MIRI group. PVN: paraventricular nucleus; MIRI: myocardial ischemia-reperfusion injury; EA: electroacupuncture.

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