Efficacy of needling Baihui (GV20), Neiguan (PC6), Shenmen (HT7) and Taichong (LR3) on cerebral cortical blood oxygen level in rats with insomnia

doi:10.19852/j.cnki.jtcm.20230404.005

Abstract

Abstract:

OBJECTIVE: To evaluate the effect of needling Baihui (GV20), Neiguan (PC6), Shenmen (HT7) and Taichong (LR3) on cerebral cortical blood oxygen level in rats with insomnia identified as liver-Qi stagnation pattern in terms of Traditional Chinese Medicine.

METHODS: Sixty Wistar rats were randomly divided into blank group (n = 10), the rest of the rats were clamped tail as stimulation combined with injected intraperitoneally p-chlorophenylalanine (PCPA) to establish sleep deprivation model. After the successful replication of the model, they were randomly divided into model group, grabbing group, Western Medicine group, acupuncture group and sham acupuncture group, with 10 rats in each group. The model group was given normal saline; The grasping group was grasped in the same way as the two treatment groups; Western medicine group was given estazolam solution; The acupuncture group was treated with the acupuncture method of "soothing the liver Qi and regulating mind" needling Baihui (GV20), Neiguan (PC6), Shenmen (HT7) and Taichong (LR3); The sham acupuncture group were needled with four non-acupoint sites. After 7 d of treatment of rats in each group, sodium pentobarbital sleep experiment was used to evaluate the sleep latency (SL) and sleep time (ST). The percentage of rats entering the open arm (OE%) and the percentage of time entering the open arm (OT%) in each group were monitored by elevated cross maze, and the vertical score, horizontal crossing times, central grid score and modification times were monitored by open field test, functional near infrared spectroscopy (fNIRs) was used to monitor the changes of oxygenated hemoglobin (Oxy-Hb), deoxyhemoglobin (Deoxy-Hb) and total hemoglobin (Total-Hb) in the cerebral cortex of rats in each group under light stimulation and dark stimulation, the statistically significant channel combinations were selected from 8 light sources and 12 detectors (S-D). According to the position of the light source detector covering the cerebral cortex, Preliminarily determine the key brain areas of insomnia (the preliminary experiment found that 6S-8D and 7S-9D were the key channels of insomnia under light stimulation, and the corresponding brain areas were prefrontal lobe and occipital lobe respectively; 7S-7D channel under dark stimulation, and the corresponding brain area was occipital lobe); The hemodynamic map of cerebral cortex was constructed through the absolute value of whole brain blood oxygen level. Further identify key brain regions for insomnia.

RESULTS: Compared with the blank group, ST, OE%, OT%, the vertical score, horizontal crossing times, central grid score, the concentration of Deoxy-Hb in prefrontal and occipital lobes were decreased significantly (P<0.01), and the concentrations of SL, modification times, Oxy-Hb and Total-Hb were increased significantly (P<0.01); There was no difference in the above indexes between the model group and the grabbing group (P>0.05); After treatment, ST, OE%, OT%, the vertical score, horizontal crossing times, central grid score and Deoxy-Hb concentration in acupuncture group and Western medicine group increased significantly, while SL, modification times, Oxy-Hb and Total-Hb concentrations were decreased significantly (P<0.01, P<0.05), Compared with the Western Medicine group, the values of OE% and OT% were more significantly higher in the acupuncture group (P<0.05), while the rest of the indexes were not significantly different between the two groups (P>0.05); Compared with the acupuncture group, ST, OE%, OT%, the vertical score, horizontal crossing times, central grid score and the concentration of deoxy Hb in the sham acupuncture group decreased significantly (P<0.01), and the concentrations of SL, modification times, Oxy-Hb and Total-Hb increased significantly (P<0.01).

CONCLUSIONS: The needling method for soothing the liver Qi and regulating mind could improve the abnormal behavior of insomnia rats with liver Qi stagnation, and its effect on the improvement of abnormal mood caused by insomnia with liver Qi stagnation is better than that of Western Medicine, and the mechanism of action may be related to the regulation of blood oxygen metabolism in the prefrontal and occipital lobes of the cerebral cortex by acupuncture.

Key words: sleep initiation and maintenance disorders, acupuncture, spectroscopy, near-infrared, hemoglobins, brain function

YE Yujuan, WEI Yuting, JIA Jing, YAN Xingke. Efficacy of needling Baihui (GV20), Neiguan (PC6), Shenmen (HT7) and Taichong (LR3) on cerebral cortical blood oxygen level in rats with insomnia[J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 523-532.

Figures/Tables 7

Table 1 Comparison of pentobarbital sodium sleep test results of rats in each group after treatment ($\bar{x}±s$, t/min)

Group	n	SL	ST
Blank	10	2.7±0.7	102.2±5.2
Model	10	4.4±0.7^a	78.1±14.2^a
Grabbing	10	4.5±0.8	74.7±2.4
Western Medicine	10	2.9±0.6^b	94.8±13.6^b
Acupuncture	10	3.0±0.5^b	88.1±3.9^b
Sham acupuncture	10	4.4±1.0^c	69.3±4.5^c

Table 2 Comparison of elevated cross maze results of rats in each group after treatment ($\bar{x}±s$)

Group	n	OE%	OT%
Blank	10	65.9±5.4	66.5±4.3
Model	10	40.0±5.3^a	46.8±3.9^a
Grabbing	10	42.1±5.3	50.0±2.7
Western Medicine	10	57.8±3.9^b	60.7±6.1^b
Acupuncture	10	64.3±2.3^bc	68.2±6.0^bc
Sham acupuncture	10	43.7±4.2^d	43.1±5.7^d

Table 3 Comparison of open field test results of rats in each group after treatment ($\bar{x}±s$)

Group	n	Vertical score	Horizontal crossing times	Center grid score	Modification times
Blank	10	11.2±1.5	26.7±4.1	19.6±1.7	1.7±0.8
Model	10	5.9±1.9^a	16.8±2.2^a	14.3±1.2^a	3.6±1.1^a
Grabbing	10	6.4±1.7	17.1±2.2	13.7±1.6	3.3±1.1
Western Medicine	10	10.9±1.4^b	24.9±1.7^b	65.7±5.5^b	1.6±1.1^b
Acupuncture	10	11.1±1.4^b	24.8±2.7^b	19.3±2.4^b	1.8±1.0^c
Sham acupuncture	10	5.5±1.3^d	16.9±2.0^d	13.4±1.5^d	3.6±1.1^d

Figure 1 Localization of brain area The blue area indicates that the cerebral cortex is inhibited, and the red area indicates that the cerebral cortex is activated. A: blood flow state of the prefrontal lobe of rats in the blank group, B: blood flow state of the prefrontal lobe of rats in the model group, C: blood flow state of the occipital lobe of rats in the blank group, D: blood flow state of the occipital lobe of rats in the model group.

Figure 2 Schematic diagram of cerebral cortical blood flow in resting state of rats in each group The closer to dark red, the higher the concentration of oxy-Hb, indicating that the activity of cerebral cortex is relatively active; The closer to dark blue, the lower the concentration of oxy-Hb, suggesting that the activity of cerebral cortex is inhibited to a certain extent. A: blank group; B: model group; C: grasping group; D: western medicine group; E: acupuncture group; F: sham acupuncture group. Blank group: rats without insomnia modeling with liver-Qi stagnation and acupuncture treatment; model group: rats with insomnia modeling with liver-Qi stagnation; grabbing group: rats with insomnia modeling with liver-Qi stagnation and grasping stimulation; Western Medicine group: rats with insomnia modeling with liver-Qi stagnation and eszopiclone treatment; acupuncture group: rats with insomnia modeling with liver-Qi stagnation and acupuncture treatment; sham acupuncture group: rats with insomnia modeling with liver-Qi stagnation and sham acupuncture treatment.

Figure 3 Determination of brain area The blue area indicates that the cerebral cortex is in an inhibited state, and the red area indicates that the cerebral cortex is in an activated state. A: blood flow state of the occipital lobe of rats in the blank group, B: blood flow state of the occipital lobe of rats in the model group. Blank group: rats without insomnia modeling with liver-Qi stagnation and acupuncture treatment; model group: rats with insomnia modeling with liver-Qi stagnation; grabbing group: rats with insomnia modeling with liver-Qi stagnation and grasping stimulation; Western Medicine group: rats with insomnia modeling with liver-Qi stagnation and eszopiclone treatment; acupuncture group: rats with insomnia modeling with liver-Qi stagnation and acupuncture treatment; sham acupuncture group: rats with insomnia modeling with liver-Qi stagnation and sham acupuncture treatment.

Figure 4 schematic diagram of cerebral cortical blood flow in resting state of rats in each group The closer to dark red, the higher the concentration of oxy Hb, indicating that the activity of cerebral cortex is relatively active; the closer to dark blue, the lower the concentration of oxy Hb, suggesting that the activity of cerebral cortex is inhibited to a certain extent. A: blank group; B: model group; C: grasping group; D: Western Medicine group; E: acupuncture group; F: sham acupuncture group. blank group: rats without insomnia modeling with liver-Qi stagnation and acupuncture treatment; model group: rats with insomnia modeling with liver-Qi stagnation; grabbing group: rats with insomnia modeling with liver-Qi stagnation and grasping stimulation; Western Medicine group: rats with insomnia modeling with liver-Qi stagnation and eszopiclone treatment; acupuncture group: rats with insomnia modeling with liver-Qi stagnation and acupuncture treatment; sham acupuncture group: rats with insomnia modeling with liver-Qi stagnation and sham acupuncture treatment.

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