Effect of electroacupuncture on the activation of microglia in rats with hypoxia-ischemia brain damage by regulating Nrf2/HO-1 pathway

doi:10.11958/20220792

Abstract

Abstract:

Objective To explore the effect of electroacupuncture regulating nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) pathway on the activation of microglia in rats with hypoxia-ischemia brain damage (HIBD). Methods Forty SD rats were randomly divided into the sham operation group, the model group, the electroacupuncture group and the electroacupuncture + all-trans retinoic acid group. Rat model of HIBD was established by common carotid artery ligation and hypoxia in the three groups except the sham operation group. In the sham operation group, only the left common carotid artery and vagus nerve were exposed no ligation and hypoxia treatment. After modeling, electroacupuncture stimulation at Baihui point was given to the electroacupuncture group. Rats in the electroacupuncture + all-trans retinoic acid group were given all-trans retinoic acid (7 mg/kg) intraperitoneally after electroacupuncture stimulation. Neurobehavioral scores of rats were evaluated by Longa score. Open field and water maze experiments were used to detect the behavioral changes and cognitive function of rats. TUNEL method was used to detect the apoptosis of nerve cells. Immunohistochemical method was used to detect the expression of microglia marker protein ionized calcium binding adapter molecule 1 (Iba1) in rat brain. Levels of interleukin-10 (IL-10), IL-1β, reactive oxygen species (ROS) and malondialdehyde (MDA) in rat brain were detected with kits. Western blot assay was used to detect the expression levels of CD68, inducible nitric oxide synthase (iNOS), arginase and Nrf2/HO-1 pathway related proteins in brain tissue. Results Compared with the sham operation group, the neurobehavioral score, the escape latency, cerebral cortex nerve cell apoptosis rate, the expression levels of Iba1, IL-1β and MDA, ROS fluorescence intensity, CD68 and iNOS were increased in the model group, and the exercise distance, number of standing times, the number of crossing platform times, levels of IL-10, arginase, Nrf2 and HO-1 decreased (P<0.05). Electroacupuncture can reduce neurobehavioral score, the escape latency, nerve cell apoptosis rate, expression levels of Iba1, IL-1β, MDA, ROS fluorescence intensity, CD68 and iNOS, and increase exercise distance, number of standing times, number of crossing platform times, levels of IL-10, arginase, Nrf2 and HO-1 (P<0.05). The effect of electroacupuncture on the above indicators could be reversed in the electroacupuncture + all-trans retinoic acid group. Conclusion Electroacupuncture may inhibit the hyperactivation of microglia in rats with HIBD by activating the Nrf2/HO-1 pathway, and promote the transition from M1 type to M2 type.

Key words: hypoxia-ischemia, brain, animals, newborn, electroacupuncture, nuclear factor NF-E2-related factor, heme oxygenase 1, microglia

CLC Number:

R285.5

Figures/Tables 7

References 18

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组别		旷场实验
组别		运动路程（cm）		站立次数（次）
假手术组		2 253.30±230.41		70.16±6.03
模型组		852.17±118.80^a		42.57±3.22^a
电针组		1 695.16±161.30^ab		61.07±4.04^ab
电针+全反式维甲酸组		1 233.46±127.14^abc		48.20±3.26^abc
F		133.590^＊＊		84.406^＊＊
组别	水迷宫实验
组别	穿越平台次数（次）		逃避潜伏期（s）
假手术组	8.93±1.02		14.10±1.12
模型组	2.04±0.50^a		27.96±2.17^a
电针组	6.25±0.71^ab		18.40±1.22^ab
电针+全反式维甲酸组	4.37±0.60^abc		22.70±1.46^abc
F	158.019^＊＊		146.818^＊＊

组别		旷场实验
组别		运动路程（cm）		站立次数（次）
假手术组		2 253.30±230.41		70.16±6.03
模型组		852.17±118.80^a		42.57±3.22^a
电针组		1 695.16±161.30^ab		61.07±4.04^ab
电针+全反式维甲酸组		1 233.46±127.14^abc		48.20±3.26^abc
F		133.590^＊＊		84.406^＊＊
组别	水迷宫实验
组别	穿越平台次数（次）		逃避潜伏期（s）
假手术组	8.93±1.02		14.10±1.12
模型组	2.04±0.50^a		27.96±2.17^a
电针组	6.25±0.71^ab		18.40±1.22^ab
电针+全反式维甲酸组	4.37±0.60^abc		22.70±1.46^abc
F	158.019^＊＊		146.818^＊＊

组别	细胞凋亡率（%）	Iba1阳性细胞数（个/视野）
假手术组	7.21±1.18	4.12±1.02
模型组	39.16±2.22^a	89.87±9.08^a
电针组	18.69±1.10^ab	40.64±5.36^ab
电针+全反式维甲酸组	23.64±2.25^abc	68.18±6.37^abc
F	278.843^＊＊	179.359^＊＊

组别	细胞凋亡率（%）	Iba1阳性细胞数（个/视野）
假手术组	7.21±1.18	4.12±1.02
模型组	39.16±2.22^a	89.87±9.08^a
电针组	18.69±1.10^ab	40.64±5.36^ab
电针+全反式维甲酸组	23.64±2.25^abc	68.18±6.37^abc
F	278.843^＊＊	179.359^＊＊

组别	IL-1β（mg/g）	IL-10（mg/g）	ROS荧光强度	MDA（mmol/mg）
假手术组	0.015±0.004	0.053±0.004	125.59±18.36	3.98±0.28
模型组	0.050±0.007^a	0.018±0.003^a	284.74±25.87^a	6.67±0.32^a
电针组	0.023±0.005^b	0.042±0.005^ab	180.15±15.36^ab	5.01±0.29^ab
电针+全反式维甲酸组	0.038±0.005^abc	0.030±0.004^abc	227.15±16.50^abc	5.80±0.30^abc
F	42.261^＊＊	69.167^＊＊	60.618^＊＊	73.946^＊＊