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网络药理学探讨野黄芩苷治疗脑缺血的作用机制及对 JAK2/STAT3 信号通路的实验验证。

Network pharmacology to explore the mechanism of scutellarin in the treatment of brain ischaemia and experimental verification of JAK2/STAT3 signalling pathway.

机构信息

Department of Anatomy and Histology/Embryology, School of Basic Medical Sciences, Kunming Medical University, 1168 West Chunrong Road, Kunming, 650500, People's Republic of China.

出版信息

Sci Rep. 2023 May 9;13(1):7557. doi: 10.1038/s41598-023-33156-5.

DOI:10.1038/s41598-023-33156-5
PMID:37160937
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10169761/
Abstract

Scutellarin is used to treat brain ischaemia. However, its underlying mechanism of action remains unclear. This study aimed to elucidate the potential mechanism of action of scutellarin in brain ischaemia through network pharmacology and experimental verification. The JAK2/STAT3 signalling pathway was identified and experimentally verified. Expression of JAK2/STAT3 signalling related proteins in TNC-1 astrocytes with BV-2 microglia-conditioned medium (CM), CM + lipopolysaccharide (LPS) (CM + L), and CM pretreated with scutellarin + LPS (CM + SL) was analysed by Western Blot and immunofluorescence staining. Expression levels of JAK2, p-JAK2, STAT3, and p-STAT3 were evaluated in astrocytes pre-treated with AG490. Middle cerebral artery occlusion (MCAO) in rats was performed in different experimental groups to detect expression of the above biomarkers. Network pharmacology suggested that the JAK2/STAT3 signalling pathway is one of the mechanisms by which scutellarin mitigates cerebral ischaemic damage. In TNC-1 astrocytes, p-JAK2 and p-STAT3 expression were significantly up-regulated in the CM + L group. Scutellarin promoted the up-regulation of various markers and AG490 neutralised the effect of scutellarin. In vivo, up-regulation of p-JAK2 and p-STAT3 after ischaemia is known. These results are consistent with previous reports. Scutellarin further enhanced this upregulation at 1, 3, and 7 d after MCAO. Scutellarin exerts its therapeutic effects on cerebral ischaemia by activating the astrocyte JAK2/STAT3 signalling, which provides a firm experimental basis for its clinical application.

摘要

野黄芩苷可用于治疗脑缺血。然而，其作用机制尚不清楚。本研究旨在通过网络药理学和实验验证来阐明野黄芩苷治疗脑缺血的潜在作用机制。确定了 JAK2/STAT3 信号通路，并进行了实验验证。通过 Western Blot 和免疫荧光染色分析 TNC-1 星形胶质细胞中 JAK2/STAT3 信号相关蛋白在 BV-2 小胶质细胞条件培养基（CM）、CM+脂多糖（LPS）（CM+L）和野黄芩苷预处理的 CM+LPS（CM+SL）中的表达。用 AG490 预处理星形胶质细胞，评估 JAK2、p-JAK2、STAT3 和 p-STAT3 的表达水平。在不同实验组的大鼠大脑中动脉闭塞（MCAO）中检测上述生物标志物的表达。网络药理学表明 JAK2/STAT3 信号通路是野黄芩苷减轻脑缺血损伤的机制之一。在 TNC-1 星形胶质细胞中，CM+L 组中 p-JAK2 和 p-STAT3 的表达明显上调。野黄芩苷促进了各种标志物的上调，而 AG490 中和了野黄芩苷的作用。在体内，缺血后 p-JAK2 和 p-STAT3 的上调是已知的。这些结果与之前的报道一致。野黄芩苷在 MCAO 后 1、3 和 7 天进一步增强了这种上调。野黄芩苷通过激活星形胶质细胞 JAK2/STAT3 信号发挥其对脑缺血的治疗作用，为其临床应用提供了坚实的实验基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/fd8758e67f4f/41598_2023_33156_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/479c68ce4f2c/41598_2023_33156_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/f386df357e7a/41598_2023_33156_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/b908d5c04eea/41598_2023_33156_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/a7dc3240d34d/41598_2023_33156_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/b648596d3535/41598_2023_33156_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/fd8758e67f4f/41598_2023_33156_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/479c68ce4f2c/41598_2023_33156_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/65d71dd0818a/41598_2023_33156_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/57cd5b81003f/41598_2023_33156_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/f386df357e7a/41598_2023_33156_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/b908d5c04eea/41598_2023_33156_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/a7dc3240d34d/41598_2023_33156_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/b648596d3535/41598_2023_33156_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/90c3/10169761/fd8758e67f4f/41598_2023_33156_Fig8_HTML.jpg

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