北京中医药大学东直门医院,北京市东城区海运仓5号,100700
muxiaohong@163.com
收稿:2024-12-09,
修回:2025-02-19,
纸质出版:2025-05-17
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于睿钦,莫岩君,张厚君等.舒筋健脑方对脑性瘫痪模型大鼠脑组织神经修复及PI3K-AKT-mTOR通路的影响[J].中医杂志,2025,66(10):1038-1045.
YU Ruiqin,MO Yanjun,ZHANG Houjun,et al.Effects of Shujin Jiannao Formula (舒筋健脑方) on Neural Repair and PI3K-Akt-mTOR Pathway of Brain Tissue in Cerebral Palsy Model Rats[J].Journal of Traditional Chinese Medicine,2025,66(10):1038-1045.
于睿钦,莫岩君,张厚君等.舒筋健脑方对脑性瘫痪模型大鼠脑组织神经修复及PI3K-AKT-mTOR通路的影响[J].中医杂志,2025,66(10):1038-1045. DOI: 10.13288/j.11-2166/r.2025.10.011.
YU Ruiqin,MO Yanjun,ZHANG Houjun,et al.Effects of Shujin Jiannao Formula (舒筋健脑方) on Neural Repair and PI3K-Akt-mTOR Pathway of Brain Tissue in Cerebral Palsy Model Rats[J].Journal of Traditional Chinese Medicine,2025,66(10):1038-1045. DOI: 10.13288/j.11-2166/r.2025.10.011.
目的
2
探讨舒筋健脑方治疗脑性瘫痪的可能作用机制。
方法
2
将30只7日龄SD大鼠随机分为正常组、模型组和舒筋健脑方组,每组10只。模型组与舒筋健脑方组采用经典Rice-Vannucci模型方法建立脑性瘫痪模型。造模成功后舒筋健脑方组大鼠予舒筋健脑方16 g/(kg·d)灌胃,正常组与模型组予生理盐水10 ml/(kg·d)灌胃,每日1次。干预1周后检测大鼠体质量,进行Zea-Longa评分及翻身实验、后肢悬吊实验评估;HE染色和尼氏染色观察脑组织病理形态,并计算尼氏染色阳性神经元数量;ELISA法检测脑组织炎症因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)含量;免疫荧光法检测脑组织神经丝蛋白200(NF200)、髓鞘碱性蛋白(MBP)水平;Western Blot法检测脑组织磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)蛋白水平。
结果
2
与正常组比较,模型组大鼠Zea-Longa评分升高,后肢悬吊实验评分降低(
P
<0.01);病理结果显示脑组织大脑皮层结构松散、海马萎缩,脑细胞损伤,IL-1β、TNF-α含量升高,皮层及海马CA1区尼氏染色阳性神经元数量减少,NF200、MBP免疫荧光强度及PI3K、mTOR蛋白水平均降低(
P
<0.05或
P
<0.01)。与模型
组比较,舒筋健脑方组大鼠Zea-Longa评分降低,后肢悬吊实验评分升高(
P
<0.05);舒筋健脑方组脑组织病理损伤有所改善,IL-1β、TNF-α含量降低,皮层及海马CA1区尼氏染色阳性神经元数量增多,NF200、MBP免疫荧光强度及PI3K、mTOR蛋白水平均升高(
P
<0.05或
P
<0.01)。各组大鼠体质量、翻身时间、脑组织AKT蛋白水平比较差异均无统计学意义(
P
>0.05)。
结论
2
舒筋健脑方可改善脑性瘫痪模型大鼠的神经功能,发挥神经修复作用,其作用机制可能与降低脑组织炎症反应和激活PI3K/AKT/mTOR信号通路有关。
Objective
2
To explore the possible mechanisms of
Shujin Jiannao Formula
(舒筋健脑方) for cerebral palsy.
Methods
2
Thirty 7-day-old SD rats were randomly divided into normal group, model group, and
Shujin Jiannao Formula
group, with 10 rats in each group. The model group and
Shujin Jiannao Formula
group established a cerebral palsy model by the classic Rice-Vannucci method. After successful modeling, rats in
Shujin Jiannao Formula
group were given
Shujin Jiannao Formula
16 g/(kg·d) by gavage, while the normal group and model group were given normal saline 10 ml/(kg·d) by gavage once a day. After one week of intervention, the rats' body weight was measured, and Zea-Longa scores, the righting reflex test, and the hindlimb suspension test were conducted for assessment; hematoxylin-eosin (HE) staining and Nissl staining were used to observe pathological changes in brain tissue, and the number of Nissl-positive neurons was counted; enzyme-linked immunosorbent assay (ELISA) was employed to measure levels of inflammatory cytokines in the brain tissue, specifically interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α); immunofluorescence was used to detect the expression levels of neurofilament protein 200 (NF200) and myelin basic protein (MBP) in brain tissue; Western Blot analysis was conducted to determine the protein levels of phosphoinositide 3-kinase (PI3K), protein kinase B (Akt/PKB/Rac), and mammalian target of rapamycin (mTOR) in brain tissue.
Results
2
Compared with the normal group, rats in the model group showed significantly higher Zea-Longa scores and lower scores in the hindlimb suspension test (
P
<0.01); pathological findings revealed loose structure in the cerebral cortex, hippocampal atrophy, and neuronal damage in brain tissue. Levels of IL-1β and TNF-α elevated, and the number of Nissl-stained positive neurons in the cortex and hippocampal CA1 region reduced, and immunofluorescence intensity of NF200 and MBP, as well as protein expression levels of PI3K and mTOR, significantly decreased (
P
<0.05 or
P
<0.01). Compared with the model group, rats in
Shujin Jiannao Formula
group showed decreased Zea-Longa scores and increased hindlimb suspension test scores (
P
<0.05); pathological damage in brain tissue alleviated, levels of IL-1β and TNF-α reduced, the number of Nissl-stained positive neurons in the cortex and hippocampal CA1 region increased, and the immunofluorescence intensity of NF200 and MBP, as well as the protein levels of PI3K and mTOR, significantly elevated (
P
<0.05 or
P
<0.01). There were no statistically significant differences among the groups in body weight, body-turning time, or AKT protein levels in brain tissue (
P
>0.05).
Conclusion
2
Shujin Jiannao Formula
can improve the neurological function of rats with cerebral palsy, exert neurorestorative effects, and its mechanism of action may be related to the reduction of inflammatory response in brain tissue and the activation of the PI3K/AKT/mTOR signaling pathway.
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