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电针对脊髓损伤大鼠脊髓组织中线粒体融合及神经干细胞增殖分化的影响。

Effect of electroacupuncture of Governor Vessel on mitochondrial fusion and proliferation and differentiation of neural stem cells in spinal cord injury rats.

机构信息

The First Clinical Medical College of Henan University of Chinese Medicine, Zhengzhou 450000, China.

The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000.

出版信息

Zhen Ci Yan Jiu. 2024 Feb 25;49(2):119-126. doi: 10.13702/j.1000-0607.20230375.

Abstract

OBJECTIVES

To observe the effect of electroacupuncture (EA) at "Dazhui" (GV14) and "Jizhong"(GV6) of the Governor Vessel (GV) on mitochondrial fusion and neural stem cell (NSC) proliferation and differentiation in the spinal cord of rats with spinal cord injury (SCI), so as to investigate its mechanisms underlying improvement of SCI.

METHODS

SD rats were randomly divided into sham operation, model and EA groups, with 15 rats in each group. The SCI model was established by using a precision impactor. EA (20 Hz/100 Hz, 1-2 mA) was applied to GV14 and GV6 for 30 min, once daily for 14 days. The rats' hindlimb locomotor function in each group was assessed using the Basso-Beattie-Bresnahan (BBB) locomotor scale. Histopathological changes of the injured spinal cord tissue and the number of neurons were evaluated after H.E. staining and Nissl staining. The expressions of Nestin, mitochondrial fusion-related protein optic atrophy-1 (OPA1) and NSC markers sex-determining region Y-box 2 (SOX2) in the injured spinal cord tissue were detected by immunofluorescence staining. The protein and mRNA expression levels of Nestin in the spinal cord tissue were detected by quantitative real-time PCR and Western blot, separately.

RESULTS

Compared with the sham operation group, the BBB scores after modeling, and the number of neurons were significantly decreased (0.001), while the mean fluorescence intensity values of Nestin, SOX2 and OPA1, and the expressions of Nestin mRNA and protein considerably increased (0.001, 0.01, 0.05) in the model group. After EA intervention and in comparison with the model group, the BBB scores at the 7 and 14 day, the number of neurons, the mean fluorescence intensity values of Nestin, SOX2 and OPA1, and the expressions of Nestin mRNA and protein were strikingly increased (0.05, 0.01, 0.001) in the EA group. H.E. staining showed swollen, ruptured and necrotic neurons of the spinal cord, with a large number of vacuoles and severe inflammatory cell infiltration after modeling, which was relatively milder in the EA group.

CONCLUSIONS

EA stimulation of GV14 and GV6 can promote the recovery of motor function in rats with SCI, which may be related to its effects in promoting mitochondrial fusion and enhancing the proliferation and differentiation of NSCs.

摘要

目的

观察电针督脉穴(GV14)和夹脊穴(GV6)对脊髓损伤(SCI)大鼠脊髓中线粒体融合和神经干细胞(NSC)增殖分化的影响,探讨其改善 SCI 的机制。

方法

将 SD 大鼠随机分为假手术组、模型组和电针组,每组 15 只。采用精密打击器建立 SCI 模型。电针(20 Hz/100 Hz,1-2 mA)每天治疗 1 次,每次 30 min,连续治疗 14 天。采用 Basso-Beattie-Bresnahan(BBB)运动评分评估各组大鼠后肢运动功能。HE 染色和尼氏染色评估损伤脊髓组织的组织病理学变化和神经元数量。免疫荧光染色检测损伤脊髓组织中巢蛋白、线粒体融合相关蛋白视神经萎缩 1(OPA1)和 NSC 标志物性别决定区 Y 框 2(SOX2)的表达。实时定量 PCR 和 Western blot 分别检测脊髓组织中巢蛋白的蛋白和 mRNA 表达水平。

结果

与假手术组相比,模型组造模后 BBB 评分和神经元数量明显减少(0.001),而巢蛋白、SOX2 和 OPA1 的平均荧光强度值以及巢蛋白 mRNA 和蛋白的表达明显增加(0.001,0.01,0.05)。电针干预后,与模型组相比,7 天和 14 天的 BBB 评分、神经元数量、巢蛋白、SOX2 和 OPA1 的平均荧光强度值以及巢蛋白 mRNA 和蛋白的表达明显增加(0.05,0.01,0.001)。电针组。HE 染色显示脊髓神经元肿胀、破裂和坏死,大量空泡形成,炎症细胞浸润严重,电针组较轻。

结论

电针刺激 GV14 和 GV6 可促进 SCI 大鼠运动功能恢复,可能与其促进线粒体融合、增强 NSCs 增殖分化有关。

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