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Astragaloside IV alleviates Schwann cell injury in diabetic peripheral neuropathy by regulating microRNA-155-mediated autophagy
Phytomedicine ( IF 7.9 ) Pub Date : 2021-09-16 , DOI: 10.1016/j.phymed.2021.153749
Yundong Yin 1 , Hua Qu 2 , Qiaoning Yang 2 , Zhaohui Fang 3 , Rui Gao 2
Affiliation  

Background

MicroRNA-155(miR-155) is closely associated with diabetic peripheral neuropathy (DPN). Astragaloside IV (AST) is a significant extract of Astragalus membranaceus, which has been found to be effective in the treatment of DPN. However, whether astragaloside IV alleviate DPN via regulating miR-155-mediated autophagy remains unclear.

Purpose

This study was designed to evaluate the effects of AST on DPN myelin Schwann cells injury and explore the mechanism of AST in treating DPN for the first time.

Methods

GK rats fed with high-fat diet and RSC96 cells cultured in high glucose were used to establish DPN Schwann cells injury in vivo and in vitro model. The effects of AST on DPN were explored through blood glucose detection, nerve function detection, pathological detection and the expression of Neuritin detected by immunohistochemical. To study the effect of AST on the DPN Schwann cells autophagy and the upstream PI3K/Akt/mTOR pathway, the expressions of beclin-1 and LC3 were detected by western blot (WB) in sciatic nerves and by immunofluorescence (IFC) in RSC96 cells. The real-time polymerase chain reaction (RT-PCR) was applied to detect the expressions of miR-155, ATG5, ATG12 both in vivo and in vitro. The binding effect of miR-155 and target gene PI3KCA was verified by luciferase reporter gene assay. The expressions of PI3K, p-Akt/Akt, p-mTOR/mTOR were detected by WB and the expressions of PI3KCA were detected by RT-PCR in vitro. The apoptosis was detected by flow cytometry. Meanwhile, the influence of miR-155 overexpression and knocked down on the above indicators was also detected in RSC96 cells. At last, further mechanism experiments were conducted to verify the mechanism of AST regulating the autophagy and apoptosis of RSC96 cells.

Results

AST reduced blood glucose levels, alleviated peripheral nerve myelin sheath injury, and improved neurological function in DPN rats. In addition, AST enhanced the autophagy activity and alleviated the apoptosis in RSC96 cell. Mechanism study shown that AST promote autophagy via regulating miR-155-mediated PI3K/Akt/mTOR signaling pathways. AST reduced RSC96 cells apoptosis by promoting autophagy.

Conclusion

AST alleviate the myelin sheath injury of DPN caused by the apoptosis of Schwann cells via enhancing autophagy, which was attributed to inhibiting the activation of the PI3K/Akt/mTOR signaling pathway by upregulating miR-155 expression.



中文翻译:

黄芪甲苷通过调节 microRNA-155 介导的自噬减轻糖尿病周围神经病变的雪旺氏细胞损伤

背景

MicroRNA-155(miR-155) 与糖尿病周围神经病变 (DPN) 密切相关。黄芪甲苷 (AST) 是黄芪的重要提取物,已被发现可有效治疗 DPN。然而,黄芪甲苷是否通过调节 miR-155 介导的自噬减轻 DPN 仍不清楚。

目的

本研究旨在评估AST对DPN髓鞘雪旺细胞损伤的影响,并首次探讨AST治疗DPN的机制。

方法

采用高脂饮食和高糖培养RSC96细胞的GK大鼠建立体内外DPN雪旺细胞损伤模型。通过血糖检测、神经功能检测、病理检测以及免疫组化检测Neuritin的表达,探讨AST对DPN的影响。为研究AST对DPN雪旺细胞自噬及上游PI3K/Akt/mTOR通路的影响,采用western blot(WB)和免疫荧光(IFC)检测坐骨神经中beclin-1和LC3的表达。 . 应用实时聚合酶链反应(RT-PCR)检测体内miR-155、ATG5 、 ATG12的表达. 荧光素酶报告基因检测验证了miR-155与靶基因PI3KCA的结合作用。WB检测PI3K、p-Akt/Akt、p-mTOR/mTOR的表达,体外RT-PCR检测PI3KCA的表达。流式细胞仪检测细胞凋亡。同时,在RSC96细胞中也检测到miR-155过表达和敲低对上述指标的影响。最后,进一步的机制实验验证了AST调节RSC96细胞自噬和凋亡的机制。

结果

AST 降低了 DPN 大鼠的血糖水平,减轻了周围神经髓鞘损伤,并改善了神经功能。此外,AST增强了RSC96细胞的自噬活性并减轻了细胞凋亡。机制研究表明,AST通过调节miR-155介导的PI3K/Akt/mTOR信号通路促进自噬。AST 通过促进自噬减少 RSC96 细胞凋亡。

结论

AST通过增强自噬来减轻雪旺细胞凋亡引起的DPN髓鞘损伤,这与通过上调miR-155表达抑制PI3K/Akt/mTOR信号通路的激活有关。

更新日期:2021-09-30
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