The mechanism of Du meridian electroacupuncture regulating cystine/glutamate reverse transporter to improve limb spasm after stroke

doi:10.11958/20231330

Abstract

Abstract:

Objective To investigate the effect and mechanism of Du meridian electroacupuncture on cystine/glutamate reverse transporter [System Xc (-)] in cerebral cortex of middle cerebral artery occlusion/reperfusion (MCAO/R) model rats. Methods A total of 60 SPF male Sprague-Dawley (SD) rats were randomly divided into the blank group, the sham operation group and the model group. The blank group did not do any treatment, the sham operation group only separated the blood vessels and the model group was prepared by middle cerebral artery embolization for MCAO/R rat model. On the third day after operation, Zea-Longa neurological deficit score, muscle tension evaluation and electrophysiological tracing were used to screen rats with limb spasm. After successful modeling, rats was randomly divided into the model group and the Du meridian electroacupuncture group. The Du meridian electroacupuncture group began treatment on the third day after operation, once a day for 7 days. After the end of treatment, the neurological function and muscle tension recovery were evaluated again. TTC staining was used to measure the cerebral infarction volume. The brain water content of rats in each group was detected by wet and dry weight method. The concentrations of glutamic acid (Glu), cysteine (Cys) and glutathione (GSH) in cerebral cortex were detected by colorimetric assay kit. Western blot assay and quantitative real-time PCR (RT-qPCR) were used to detect the expression of SLC7A11, SLC3A2, γ-glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GSS) in cerebral cortex at the protein level and gene level, respectively. Results Compared with the model group, the neurological deficit score and modified Ashworth score were decreased in the Du meridian electroacupuncture group, the EMG signal value was enhanced, the brain water content was decreased, the degree of cerebral infarction was restored, the contents of Cys and GSH were significantly increased, the expression levels of SLC7A11, SLC3A2, γ-GCS and GSS protein and mRNA were significantly increased, and the concentration of Glu was significantly decreased (P<0.05). Conclusion Du meridian electroacupuncture can improve the degree of neurological impairment and limb spasm in rats after stroke. The mechanism may be related to the regulation of System Xc (-) and improving oxidative stress.

Key words: stroke, spasm, du meridian electroacupuncture, System Xc (-)

CLC Number:

R743.3

Figures/Tables 9

References 24

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基因名称	引物序列（5′→3′）	产物大小/ bp
SLC7A11	上游：TCCCGGCGTATTACCTCTTC	122
SLC7A11	下游：TTCTTTAGAGTCTTCTGGTACAACT	122
SLC3A2	上游：CGAGAGGCATAGCTGGTCTG	74
SLC3A2	下游：GCCCAAAACTAGGCCCTTCA	74
γ-GCS	上游：TCAGTCCACAAACTGGCAGA	90
γ-GCS	下游：ACCTCCATTGGTCGGAACTC	90
GSS	上游：ATCTTGCAGGATGAGAAGCAGC	102
GSS	下游：GGAGCTGGGGTTCTTTGCG	102
GAPDH	上游：ACAGCAACAGGGTGGTGGAC	252
GAPDH	下游：TTTGAGGGTGCAGCGAACTT	252

基因名称	引物序列（5′→3′）	产物大小/ bp
SLC7A11	上游：TCCCGGCGTATTACCTCTTC	122
SLC7A11	下游：TTCTTTAGAGTCTTCTGGTACAACT	122
SLC3A2	上游：CGAGAGGCATAGCTGGTCTG	74
SLC3A2	下游：GCCCAAAACTAGGCCCTTCA	74
γ-GCS	上游：TCAGTCCACAAACTGGCAGA	90
γ-GCS	下游：ACCTCCATTGGTCGGAACTC	90
GSS	上游：ATCTTGCAGGATGAGAAGCAGC	102
GSS	下游：GGAGCTGGGGTTCTTTGCG	102
GAPDH	上游：ACAGCAACAGGGTGGTGGAC	252
GAPDH	下游：TTTGAGGGTGCAGCGAACTT	252

组别	干预前	干预后	t
空白组	0±0	0±0	-
假手术组	0±0	0±0	-
模型组	2.08±0.67^ab	1.92±0.67^ab	0.611
督脉电针组	2.00±0.60^ab	0.50±0.67^c	5.745^＊
F	82.300^＊	43.700^＊

组别	干预前	干预后	t
空白组	0±0	0±0	-
假手术组	0±0	0±0	-
模型组	2.08±0.67^ab	1.92±0.67^ab	0.611
督脉电针组	2.00±0.60^ab	0.50±0.67^c	5.745^＊
F	82.300^＊	43.700^＊

组别	干预前	干预后	t
空白组	0±0	0±0	-
假手术组	0±0	0±0	-
模型组	2.42±0.67^ab	2.17±0.72^ab	0.573
督脉电针组	2.25±0.75^ab	0.42±0.67^c	6.303^＊
F	86.000^＊	53.200^＊