1.中山大学附属第一医院针灸科,广东 广州510080
2.中山大学中山医学院,广东 广州510080
焦博钰,硕士生,研究方向:电针对睡眠剥夺的机制研究,E-mail:97986575@qq.com
纸质出版日期:2022-11-20,
收稿日期:2022-07-19,
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焦博钰,孟凡琪,肖兴茹等.四神聪电针预防急性睡眠剥夺大鼠海马神经元凋亡的机制及对焦虑行为的影响[J].中山大学学报(医学科学版),2022,43(06):958-966.
JIAO Bo-yu,MENG Fan-qi,XIAO Xing-ru,et al.Mechanism of Electroacupuncture on Preventing Apoptosis of Hippocampal Neurons in Rats with Acute Sleep Deprivation and Its Effect on Anxiety[J].Journal of Sun Yat-sen University(Medical Sciences),2022,43(06):958-966.
焦博钰,孟凡琪,肖兴茹等.四神聪电针预防急性睡眠剥夺大鼠海马神经元凋亡的机制及对焦虑行为的影响[J].中山大学学报(医学科学版),2022,43(06):958-966. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2022.0611.
JIAO Bo-yu,MENG Fan-qi,XIAO Xing-ru,et al.Mechanism of Electroacupuncture on Preventing Apoptosis of Hippocampal Neurons in Rats with Acute Sleep Deprivation and Its Effect on Anxiety[J].Journal of Sun Yat-sen University(Medical Sciences),2022,43(06):958-966. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2022.0611.
目的
2
探讨四神聪电针预防急性睡眠剥夺大鼠海马神经元凋亡的机制及对焦虑行为的影响。
方法
2
将SD大鼠随机分为Sham组(对照组)、sleep deprivation(SD,睡眠剥夺组)、EA+SD组(eletroacupuncture,电针干预组)。采用改良水平台法对SD组、EA+SD组进行连续4天的急性睡眠剥夺,电针组在睡眠剥夺期间于四神聪穴位进行电针治疗,连续4 d,每天20 min。采用旷场实验检测大鼠探索能力以评估大鼠焦虑程度,并通过免疫荧光染色检测海马中CA1、CA2、CA3、DG1、DG2区域的神经元(NeuN阳性)和Cleaved Caspase-3的共标情况;采用免疫荧光标记海马中星形胶质细胞标记物胶质纤维酸性蛋白(GFAP),通过sholl analysis分析星形胶质细胞形态变化,并检测分别与补体C3
+
、S100A
+
的共标情况,以了解海马中星形胶质细胞A1、A2分型情况。
结果
2
旷场实验发现:EA+SD组大鼠在中心停留时间的时间增加,在中心区域的运动速度下降;免疫荧光染色显示:EA+SD组大鼠在海马不同区域除CA3外,其余的CA1、CA2、DG1、DG2区神经元上Cleaved caspase-3较SD组大鼠表达减少(
P
<0.05);sholl analysis发现在12 μm、15 μm、18 μm、21 μm处EA+SD组星形胶质细胞与同心圆交点数目多于SD组,在海马区A1型星形胶质细胞(C3
+
GFAP
+
/GFAP
+
)比例低于SD组,A2型(S100A
+
GFAP
+
/GFAP
+
)所占比例高于SD组。
结论
2
四神聪电针对急性睡眠剥夺大鼠海马区神经元具有保护作用,其机制可能通过预防星形胶质细胞形态损伤,减少海马区A1型星形胶质细胞,增加A2型星形胶质细胞表达,降低急性睡眠剥夺导致的细胞毒性,减少神经元凋亡数而实现;同时,四神聪电针还能预防或降低急性睡眠剥夺对大鼠探索性行为的影响。
Objective
2
To investigate the mechanism of Sishencong electroacupuncture on preventing the apoptosis of hippocampal neurons in rats with acute sleep deprivation and its effect on anxiety behavior.
Methods
2
Adult Sprague-Dawley (SD) rats were randomly divided into Sham group (control group), SD group (sleep deprivation group), and EA+SD group (eletroacupuncture group). Modified multiple platform method was used to induce sleep deprivation for 4 days. EA+SD group received Sishencong electroacupuncture in each day during sleep deprivation with 20 minutes needling for 4 days. The open field test was used to detect the exploration of rats to evaluate the degree of anxiety, and the co-labeling of NeuN
+
and Cleaved Caspase-3 in different regions of the hippocampus was detected by immunofluorescence staining; the astrocyte marker glial fibrillary acidic protein (GFAP) in the hippocampus was labeled by immunofluorescence. GFAP and the morphological changes of astrocytes were analyzed by Sholl analysis, and the co-labeling with C3+ and S100A+ were detected to analyze the A1 and A2 types of astrocytes in the hippocampus.
Results
2
Open field test showed that the time spent in the center of rats was increased in EA+SD group, but the movement speed was decreased compared with SD group. Immunofluorescence staining confirmed that the expression of Cleaved caspase-3 on neurons in different regions of the hippocampus (CA1, CA2, CA3, DG1, DG2) in the EA+SD group was lower than that of the SD group (
P
<
0.05). The number of intersections of astrocytes and concentric circles in the EA+SD group at 12 μm,15 μm, 18 μm and 21 μm were higher than those of the SD group. Immunofluorescence showed that electroacupuncture could regulate the proportion of C3
+
/GFAP
+
and S100A
+
/GFAP
+
in the hippocampus of rats during sleep deprivation.
Conclusion
2
Electroacupuncture could improve the exploration during sleep deprivation by reducing the apoptosis of neurons in the hippocampus, repairing the morphological damage of astrocytes and balancing the proportion of A1 and A2 astrocytes in the hippocampus.
睡眠剥夺电针神经元凋亡星形胶质细胞旷场实验
sleep deprivationelectroacupunctureneuronal apoptosisastrocytesopen field test
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