人参皂苷 Rg1 通过 lncRNA-Malat1/miR-124-3p/Lamc1 轴调控星形胶质细胞的激活，促进脊髓损伤修复。

Ginsenoside Rg1 Regulates the Activation of Astrocytes Through lncRNA-Malat1/miR-124-3p/Lamc1 Axis Driving PI3K/AKT Signaling Pathway, Promoting the Repair of Spinal Cord Injury.

机构信息

Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, China.

Department of Orthopedics, The Affiliated Zhangjiagang Hospital of Soochow University, Zhangjiagang, China.

出版信息

CNS Neurosci Ther. 2024 Nov;30(11):e70103. doi: 10.1111/cns.70103.

DOI:10.1111/cns.70103

PMID:39491316

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11532020/

Abstract

AIM

To investigate the regulation of ginsenoside Rg1 on the PI3K/AKT pathway through the lncRNA-Malat1/miR-124-3p/ Laminin gamma1 (Lamc1) axis, activating astrocytes (As) to promote the repair of spinal cord injury (SCI).

METHODS

Bioinformatics analysis was used to predict miRNA targeting Lamc1 and lncRNA targeting miR-124-3p, which were then validated through a dual-luciferase assay. Following transfection, the relationships between Malat1, miR-124-3p, and Lamc1 expression levels were assessed by qRT-PCR and Western blot (WB). Immunofluorescence staining and immunohistochemistry were utilized to measure Lamc1 expression, while changes in cavity area were observed through hematoxylin-eosin (HE) staining. Basso-Beattie-Bresnahan (BBB) scale and footprint analysis were used to evaluate functional recovery. WB was performed to assess the expression of PI3K/AKT pathway-related protein.

RESULTS

Rg1 was found to upregulate Malat1 expression, which in turn modulated the Malat1/miR-124-3p/Lamc1 axis. Furthermore, Rg1 activated the PI3K/Akt signaling pathway, significantly reducing the SCI cavity area and improving hind limb motor function. However, knockout of Malat1 hindered these effects, and inhibition of miR-124-3p reversed the silencing effects of Malat1.

CONCLUSIONS

Rg1 can induce Malat1 expression to activate the Lamc1/PI3K/AKT signaling pathway by sponging with miR-124-3p, thereby regulating As activity to repair SCI.

摘要

目的

通过长链非编码 RNA（lncRNA）-MALAT1/miR-124-3p/层粘连蛋白γ1（Lamc1）轴研究人参皂苷 Rg1 对 PI3K/AKT 通路的调节作用，激活星形胶质细胞（As）促进脊髓损伤（SCI）的修复。

方法

通过生物信息学分析预测靶向 Lamc1 的 miRNA 和靶向 miR-124-3p 的 lncRNA，然后通过双荧光素酶报告基因实验验证。转染后，通过 qRT-PCR 和 Western blot（WB）评估 Malat1、miR-124-3p 和 Lamc1 表达水平之间的关系。免疫荧光染色和免疫组织化学用于测量 Lamc1 的表达，通过苏木精-伊红（HE）染色观察腔面积的变化。Basso-Beattie-Bresnahan（BBB）评分和足迹分析用于评估功能恢复情况。WB 用于评估 PI3K/AKT 通路相关蛋白的表达。

结果

Rg1 被发现上调 Malat1 的表达，进而调节 Malat1/miR-124-3p/Lamc1 轴。此外，Rg1 激活了 PI3K/Akt 信号通路，显著减少 SCI 腔面积，改善后肢运动功能。然而，敲除 Malat1 会阻碍这些作用，抑制 miR-124-3p 逆转了 Malat1 的沉默作用。