1.宁夏医科大学 中医燥证教育部重点实验室,银川 750004
2.宁夏区域高发病中西医结合防治研究重点实验室,银川 750004
3.宁夏医科大学 中医学院,银川 750004
武润博,在读硕士,从事中医药治疗脑卒中研究,E-mail:209800866@qq.com
郭斌,教授,从事针药结合治疗疾病的临床与机制研究,E-mail:20200934@nxmu.edu.cn
收稿:2025-08-17,
修回:2025-10-30,
录用:2025-11-17,
网络首发:2025-11-20,
纸质出版:2026-05-05
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武润博,孟纯雪,王菲等.芍药甘草汤对脑卒中肢体痉挛状态大鼠神经干细胞增殖分化的影响[J].中国实验方剂学杂志,2026,32(09):11-20.
WU Runbo,MENG Chunxue,WANG Fei,et al.Effect of Shaoyao Gancaotang on Proliferation and Differentiation of Neural Stem Cells in Rats with Limb Spasticity of Cerebral Apoplexy[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(09):11-20.
武润博,孟纯雪,王菲等.芍药甘草汤对脑卒中肢体痉挛状态大鼠神经干细胞增殖分化的影响[J].中国实验方剂学杂志,2026,32(09):11-20. DOI: 10.13422/j.cnki.syfjx.20252406.
WU Runbo,MENG Chunxue,WANG Fei,et al.Effect of Shaoyao Gancaotang on Proliferation and Differentiation of Neural Stem Cells in Rats with Limb Spasticity of Cerebral Apoplexy[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(09):11-20. DOI: 10.13422/j.cnki.syfjx.20252406.
目的
2
分析脑卒中肢体痉挛状态大鼠肌张力损伤程度及大脑皮质M1区的巢蛋白(Nestin)和
β
-连环蛋白(
β
-catenin)的表达情况,探讨不同剂量芍药甘草汤对脑卒中肢体痉挛状态(SCA)大鼠的作用机制。
方法
2
通过随机分组,分为空白组,模型组,阳性药组(巴氯芬,5.25 mg·kg
-1
),芍药甘草汤低、中、高剂量组(2.1、4.2、8.4 g·kg
-1
),每组9只。采用改良光栓法+内囊注射无水乙醇法制备脑卒中肢体痉挛大鼠模型,行为学评分确认模型成功后。进行药物干预,行为学检测神经功能缺损及肌张力、旷场实验检测运动距离、透射电镜观察突触结构、骨骼肌三磷酸腺苷(ATP)酶染色分析肌纤维变化、苏木素-伊红(HE)染色观察组织形态学变化、免疫组化、实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测大脑皮质M1区中Nestin和
β
-catenin mRNA水平与蛋白表达。
结果
2
与空白组比较,模型组大鼠神经功能缺损评分显著增加(
P<
0.01),肌张力评分显著增加(
P<
0.01),矿场运动距离显著减少(
P<
0.01),突出结构肿胀模糊、大脑皮质细胞被严重破坏、骨骼肌ATP酶Ⅰ型肌纤维比例显著增加(
P
<
0.01),Nestin mRNA水平和蛋白表达显著减少(
P
<
0.01),
β
-catenin mRNA水平和蛋白表达显著增加(
P
<
0.01);与模型组比较,芍药甘草汤显著降低SCA大鼠神经功能缺损评分和肌张力评分(
P<
0.01),显著增加SCA大鼠的运动距离(
P
<
0.01);透射电镜发现芍药甘草汤组的突触结构较清晰完整,囊泡数量多且形态好转;HE染色发现芍药甘草汤组的神经细胞结构完整,排列整齐,空泡细胞数量减少;ATP酶染色结果显示芍药甘草汤组的Ⅰ型肌纤维比例减少(
P
<
0.01);Real-time PCR结果显示芍药甘草汤组显著升高大鼠Nestin和
β
-catenin mRNA表达(
P
<
0.01);免疫组化和Western blot结果表明,芍药甘草汤组明显升高SCA大鼠Nestin和
β
-catenin蛋白表达(
P
<
0.05,
P
<
0.01)。
结论
2
芍药甘草汤可能通过调节大脑皮质M1区中神经干细胞的增殖分化来改善SCA模型大鼠的神经功能损伤和肢体痉挛状态。
Objective
2
This paper aims to analyze the damage degree of muscle tone in rats with spasticity of cerebral apoplexy (SCA) and the expression of Nestin and
β
-catenin in the M1 region of the cerebral cortex, thereby investigating the action mechanism of different doses of Shaoyao Gancaotang on rats with SCA.
Methods
2
The rats were rando
mly divided into a blank group, a model group, a positive control group (baclofen, 5.25 mg·kg
-1
), and low, medium, and high-dose groups of Shaoyao Gancaotang (2.1, 4.2, 8.4 g·kg
-1
), with nine rats in each group. A rat model with SCA was established by using a modified phrenic nerve block combined with intraventricular injection of anhydrous ethanol. Following behavioral scoring to confirm model validity, drug interventions were conducted. Neurological deficits and muscle tone were evaluated by behavioral assessments. The open field test was used to measure locomotor distance. Transmission electron microscopy was employed to examine the synaptic structures. Skeletal muscle adenosine triphosphate (ATP)ase staining was used to analyze myofibrillar changes. Hematoxylin and eosin (HE) staining was used to observe the histomorphological changes. Immunohistochemistry, Real-time polymerase chain reaction (Real-time PCR), and Western blot were employed to detect mRNA levels and protein expressions of Nestin and
β
-catenin in the M1 region of the cerebral cortex.
Results
2
Compared with the blank group, rats in the model group exhibited significantly increased neurological deficit scores (
P
<
0.01), markedly elevated muscle tone scores (
P
<
0.01), substantially reduced locomotor distance (
P
<
0.01), prominent structural swelling and blurring, severe destruction of cerebral cortical cells, a significant increase in the proportion of skeletal muscle ATPase type Ⅰ fibers (
P
<
0.01), a significant decrease in mRNA levels and protein expression of Nestin (
P
<
0.01), and a significant increase in mRNA levels and protein expression of
β
-catenin (
P
<
0.01). Compared with the model group, the Shaoyao Gancaotang group exhibited reduced neurological deficit scores and muscle tone scores in rats with SCA (
P
<
0.01) and increased
locomotor distance (
P
<
0.01). Transmission electron microscopy revealed clearer and more intact synaptic structures in the rats from the Shaoyao Gancaotang group, with increased vesicle numbers and improved morphology. HE staining revealed intact neuronal cell structures with regular arrangement and reduced vacuolated cells in the rats from Shaoyao Gancaotang. ATPase staining result indicated a decreased proportion of type Ⅰ muscle fibers in the rats from the Shaoyao Gancaotang group (
P
<
0.01). Real-time PCR results demonstrated increased mRNA expressions of Nestin and
β
-catenin in the rats from the Shaoyao Gancaotang group (
P
<
0.01). Immunohistochemistry and western blot analyses indicated elevated protein expressions of Nestin and
β
-catenin in rats with SCA from the Shaoyao Gancaotang group (
P
<
0.05,
P
<
0.01).
Conclusion
2
Shaoyao Gancaotang may improve neurological function impairment and limb spasticity in model rats with SCA by regulating the proliferation and differentiation of neural stem cells in the cerebral cortex M1 region.
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