贯叶金丝桃素通过靶向 PI3K-AKT/ICAM1 轴调节肾纤维化。
Hyperforin regulates renal fibrosis via targeting the PI3K-AKT/ICAM1 axis.
机构信息
Department of Urology, The Third Xiangya Hospital of Central South University, Changsha 410013, China.
Department of Urology, The Third Xiangya Hospital of Central South University, Changsha 410013, China.
出版信息
Cell Signal. 2023 Aug;108:110691. doi: 10.1016/j.cellsig.2023.110691. Epub 2023 Apr 26.
OBJECTIVE
To explore the role and mechanism of hyperforin (one of the active components of Sophora flavescens) in renal fibrosis.
METHODS
The active compounds and target proteins of Sophora flavescens were first screened through TCMSP (https://tcmsp-e.com/). The renal fibrosis-related genes were analyzed through GeneCards (https://www.genecards.org/). The differentially expressed genes (DEGs) in renal fibrosis in GEO dataset GSE156181 were obtained. Metascape was applied for target protein enrichment analysis. TGF-β1-stimulated renal tubular epithelial cells were used for renal fibrosis cell model establishment. The unilateral ureteral obstruction (UUO) mouse model was used for the renal fibrosis in vivo model. Cell viability was detected using an MTT assay. Immunofluorescence staining was employed to detect cell morphology changes and the expression of α-SMA and collagen I. Hematoxylin and eosin (H&E) and Masson staining were employed to determine the renal morphologic change. qRT-PCR or Western blotting was applied to determine the expression levels of the target proteins.
RESULTS
After intersecting the analysis results of TCMSP, GeneCards, and dataset GSE156181, hyperforin targeting ICAM1 was identified. Metascape pathway enrichment analysis results revealed that the effective compounds of Sophora flavescens were tightly associated with extracellular matrix (ECM) remodeling and inflammatory response. MTT assay demonstrated that hyperforin had no toxic effect on cells. Immunofluorescence staining results evidenced that hyperforin could partially restore TGF-β1-induced epithelial-mesenchymal transition (EMT), the PI3K/AKT pathway activation, and ICAM1 upregulation, and these effects of hyperforin could be reversed by ICAM1 overexpression. While the PI3K/AKT pathway activator IGF-1 effectively reversed the EMT inhibition effect of hyperforin on renal tubular epithelial cells. Moreover, the UUO mouse model further confirmed that hyperforin reduced renal fibrosis.
CONCLUSION
Hyperforin inhibited renal fibrosis via the PI3K/AKT/ICAM1 axis.
目的
探讨贯叶金丝桃素（苦参的一种活性成分）在肾纤维化中的作用和机制。
方法
首先通过 TCMSP（https://tcmsp-e.com/）筛选苦参的活性化合物和靶蛋白。通过 GeneCards（https://www.genecards.org/）分析与肾纤维化相关的基因。从 GEO 数据集 GSE156181 中获取肾纤维化中差异表达的基因（DEGs）。采用 Metascape 进行靶蛋白富集分析。用 TGF-β1 刺激肾小管上皮细胞建立肾纤维化细胞模型。用单侧输尿管梗阻（UUO）小鼠模型建立肾纤维化体内模型。用 MTT 法检测细胞活力。免疫荧光染色检测细胞形态变化及α-SMA 和胶原 I 的表达。苏木精和伊红（H&E）及 Masson 染色观察肾脏形态变化。qRT-PCR 或 Western blot 检测靶蛋白的表达水平。
结果
将 TCMSP、GeneCards 和数据集 GSE156181 的分析结果进行交集分析，鉴定出贯叶金丝桃素靶向 ICAM1。Metascape 通路富集分析结果表明，苦参的有效化合物与细胞外基质（ECM）重塑和炎症反应密切相关。MTT 实验表明贯叶金丝桃素对细胞无毒作用。免疫荧光染色结果表明，贯叶金丝桃素可部分恢复 TGF-β1 诱导的上皮-间充质转化（EMT）、PI3K/AKT 通路激活和 ICAM1 上调，而过表达 ICAM1 可逆转贯叶金丝桃素的这些作用。而 PI3K/AKT 通路激活剂 IGF-1 可有效逆转贯叶金丝桃素对肾小管上皮细胞 EMT 的抑制作用。此外，UUO 小鼠模型进一步证实贯叶金丝桃素可减轻肾纤维化。
结论
贯叶金丝桃素通过 PI3K/AKT/ICAM1 轴抑制肾纤维化。
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