Effect of electroacupuncture on hypertensive and sympathetic excitability mechanism mediated by the paraventricular nucleus of the hypothalamus in spontaneous hypertensive rats

doi:10.19852/j.cnki.jtcm.2025.03.013

Abstract

Abstract:

OBJECTIVE: To investigate the mechanism of electroacupuncture of sympathetic nerve activity and blood pressure reduction in the hypothalamic paraventricular nucleus (PVN) of spontaneous hypertensive rats (SHRs).
METHODS: A total of 64 male SHRs were divided into four groups: model, sham-operated (Sham), electro-acupuncture (EA), and N-methyl-D-aspartate receptor antagonist and electroacupuncture (NRA + EA). In addition, 16 Wistar-Kyoto rats were used as controls. PVN stereotaxic surgery was performed in both the Sham and NRA + EA groups, while the EA and NRA + EA groups received 14 d of electroacupuncture. Blood pressure (BP) and heart rate (HR) were measured the day before the intervention and every other day. After 14 d of intervention, the rats in each group were tested for renal sympathetic nerve activity (RSNA). The associated factor levels were determined using Western blotting, reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assays.
RESULTS: In comparison to the model group, the EA and NRA + EA groups had significantly lower BP, HR, and RSNA (P < 0.01). The expression of N-methyl-D-aspartate receptor (NMDAR), angiotensin II (Ang II), angiotensin II type 1 (AT1), tumor necrosis factor-α, interleukin-1β, norepinephrine and arginine vasopressin was significantly lower in the EA and NRA + EA groups (P < 0.01). Moreover, the antihypertensive effect of NRA+EA group outperformed to the EA group.
CONCLUSIONS: Electroacupuncture effectively reduced the BP and sympathetic nerve excitability in SHRs. The mechanism was linked to the inhibition of NMDAR-mediated Ang II /AT1 and the inflammatory response in PVN.

Key words: electroacupuncture, hypertension, renal sympathetic nerve activity, paraventricular hypothalamic nucleus, receptors, N-methyl-D-aspartate

SUN Jiao, WANG Yueming, LYU Jian, LIU Xin, YUE Bingnan, LI Yinyin, LIU Jipeng, SUN Yize, LIU Qingguo, YAN Liu. Effect of electroacupuncture on hypertensive and sympathetic excitability mechanism mediated by the paraventricular nucleus of the hypothalamus in spontaneous hypertensive rats[J]. Journal of Traditional Chinese Medicine, 2025, 45(3): 586-596.

Figures/Tables 7

Table 1 Changes in SBP ($\bar{x}±s$)

Group	n	Day 0	Day 2	Day 4	Day 6	Day 8	Day 10	Day 12	Day 14
Control	16	108.6±1.0	109.8±0.8	111.0±1.4	112.0±1.4	112.0±1.1	113.0±0.9	112.8±1.5	113.0±0.6
Model	16	169.8±2.1^abc	170.6±1.6^abc	171.2±2.3^ab	172.2±2.4^a	173.0±1.8^a	173.6±2.3^a	173.8±1.8^a	174.4±1.9^a
Sham	16	174.8±0.8^a	174.2±1.2^a	175.0±0.9^a	175.2±0.8^a	175.8±0.8^a	174.6±1.0^a	174.2±2.1^a	174.4±1.6^a
EA	16	169.6±1.4^abc	171.2±1.2^abc	170.6±0.5^ab	168.4±1.0^abd	164.8±1.3^abd	160.4±1.0^abd	156.2±1.7^abd	152.4±2.1^abd
NRA+EA	16	174.8±0.8^a	174.8±0.8^a	174.0±1.1^a	170.4±1.0^ab	166.2±1.2^abd	160.4±1.0^abd	152.8±1.5^abd	144.8±1.7^abde

Table 2 Changes in HR ($\bar{x}±s$)

Group	n	Day 0	Day 2	Day 4	Day 6	Day 8	Day 10	Day 12	Day 14
Control	16	281.2±4.0	288.6±5.0	302.8±6.73	306.0±5.8	300.0±4.2	296.0±4.6	290.6±4.8	288.4±2.9
Model	16	350.2±3.3^a	356.0±4.4^a	368.2±5.5^a	373.8±3.4^a	383.2±5.7^a	397.2±4.0^a	409.2±5.8^a	419.0±6.2^a
Sham	16	349.6±3.1^a	360.8±1.7^a	375.2±3.7^a	385.6±4.1^a	392.8±3.8^a	398.0±3.5^a	410.4±4.3^a	420.2±2.5^a
EA	16	351.4±3.0^a	358.8±3.3^a	363.6±3.7^a	357.8±3.9^abc	350.8±3.9^abc	344.6±3.7^abc	334.6±4.8^abc	323.2±5.3^abc
NRA+EA	16	350.6±3.0^a	363.2±4.3^a	371.0±2.3^a	365.8±2.6^ab	354.8±3.6^abc	343.4±3.0^abc	329.8±4.0^abc	302.4±4.3^abcd

Figure 1 RSNA expression in different groups A: renal sympathetic nerve discharge in the control group; B: renal sympathetic nerve discharge in the model group; C: renal sympathetic nerve discharge in the sham group; D: renal sympathetic nerve discharge in the EA group; E: renal sympathetic nerve discharge in the NRA + EA group; F: renal sympathetic nerve activity (percentage relative to the maximum value). Control and Model groups: fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; Sham group: inject the Artificial cerebrospinal fluid (10 mmol/L, 100 nL) and fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; EA group: electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d; NRA + EA group: inject the N-methyl-D-aspartic acid receptor inhibitor (10 mmol/L, 100 nL) and electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d. RSNA: renal sympathetic nerve activity; EA: electroacupuncture; NRA + EA: N-methyl-D-aspartate receptor antagonist and electroacupuncture. One-way analysis of variance was adopted, and the least significant difference t-test was used for the comparison among groups. Data were presented as mean ± standard deviation (n = 16). aP < 0.01, vs control group; bP < 0.01, vs model group; cP < 0.01, vs Sham group; dP < 0.05, vs EA group.

Figure 2 NMDAR, Ang Ⅱ and AT1protein expression in each group of rats A: Western blot analysis of levels of NMDAR and Ang Ⅱ expression in PVN of rats in each group; B: comparison of the protein amount of NMDAR in each group; C: comparison of the protein amount of Ang Ⅱ in each group; D: expression of AT1 mRNA. 1: model group; 2: control group; 3: EA group; 4: sham group; 5: NRA + EA group. Control and model groups: fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; Sham group: inject the Artificial cerebrospinal fluid (10 mmol/L, 100 nL) and fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; EA group: electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d; NRA + EA group: inject the N-methyl-D-aspartic acid receptor inhibitor (10 mmol/L, 100 nL) and electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d. EA: Electroacupuncture; NRA + EA: N-methyl-D-aspartate receptor antagonist and electroacupuncture; NMDAR: N-methyl-D-aspartate receptor; Ang II: angiotensin II; AT1: angiotensin II type 1. One-way analysis of variance was adopted, and the least significant difference t-test was used for the comparison among groups. Data were presented as mean ± standard deviation (n = 16). aP < 0.01 and eP < 0.05, vs control group; bP < 0.05 and fP < 0.01, vs model group; cP < 0.05 and gP < 0.01, vs Sham group; dP < 0.05, vs EA group.

Figure 3 TNF-α levels in the PVN A: immunofluorescence images of TNF-α in different groups. A1: control group; A2: model group; A3: sham group; A4: EA group; A5: NRA+EA group; B: immunofluorescence intensity values of TNF-α in each group; C: expression of TNF-α mRNA. Control and Model groups: fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; Sham group: inject the Artificial cerebrospinal fluid (10 mmol/L, 100 nL) and fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; EA group: electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d; NRA + EA group: inject the N-methyl-D-aspartic acid receptor inhibitor (10 mmol/L, 100 nL) and electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d. EA: electroacupuncture; NRA + EA:N-methyl-D-aspartate receptor antagonist and electroacupuncture; TNF-α: tumor necrosis factor; PVN: paraventricular nucleus. One-way analysis of variance was adopted, and the least significant difference t-test was used for the comparison among groups. Data were presented as mean ± standard deviation (n = 16). aP < 0.01 and dP < 0.05, vs control group; b P < 0.05, vs model group; cP < 0.05, vs Sham group.

Figure 4 IL-1β levels in the PVN A: immunofluorescence images of IL-1β in different groups. A1: control group; A2: model group; A3: sham group; A4: EA group; A5: NRA + EA group; B: immunofluorescence intensity values of IL-1β in each group; C: expression of IL-1β mRNA. Control and Model groups: fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; Sham group: inject the Artificial cerebrospinal fluid (10 mmol/L, 100 nL) and fixed with a rat sleeve (as per rats in the EA and NRA + EA groups) without any acupuncture intervention for 14 d; EA group: electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d; NRA + EA group: inject the N-methyl-D-aspartic acid receptor inhibitor (10 mmol/L, 100 nL) and electro-acupuncture at Taichong (LR3) and Quchi (LI11) acupoints for 14 d. EA: electroacupuncture; NRA+EA: N-methyl-D-aspartate receptor antagonist and electroacupuncture; IL-1β: interleukin-1β; PVN: paraventricular nucleus. One-way analysis of variance was adopted, and the least significant difference t-test was used for the comparison among groups. Data were presented as mean ± standard deviation (n = 16). aP < 0.05, vs control group; bP < 0.05, vs model group; cP < 0.05, vs Sham group.

Table 3 NE and AVP levels in serum (pg/mL)

Group	n	NE	AVP
Control	16	1.9±0.5	1.1±0.3
Model	16	17.9±1.2^a	18.8±1.3^a
Sham	16	17.5±1.5^a	18.6±1.5^a
EA	16	7.5±0.7^bc	17.1±0.7^bc
NRA+EA	16	5.9±0.4^bc	15.4±1.1^bc

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