基于网络药理学和实验验证探究红景天调控颗粒细胞自噬的作用机制

Exploring the Mechanism of Action of Rhodiolae Crenulatae Radix et Rhizoma in Regulating Autophagy of Granulosa Cells Based on Network Pharmacology and Experimental Validation

关键词:  红景天  网络药理学  颗粒细胞  红景天苷  自噬  PI3K/AKT信号通路

Key words:  Rhodiolae Crenulatae Radix et Rhizoma  network pharmacology  granulosa cells  salidroside  autophagy  PI3K/AKT signaling pathway

摘要:

目的 基于网络药理学和实验验证探究红景天调控颗粒细胞自噬的潜在作用靶点和作用机制。方法 借助数据库获取红景天的活性成分、作用靶点以及自噬相关靶点，建立成分-靶点网络图，蛋白质互作网络图，对关键靶点进行基因和信号通路的富集分析并且对活性成分和核心靶点进行分析对接，探究红景天调控颗粒细胞自噬的可能作用机制；使用Western blotting、RT-PCR、透射电镜以及双标腺病毒检测红景天重要活性成分对颗粒细胞自噬的作用。结果 获取红景天有效成分7个，作用靶点441个，细胞自噬相关靶点796个，富集分析得到生物过程834个，细胞成分60个，分子功能69个，以及信号通路150条；分子对接结果表明红景天活性成分与核心作用靶点AKT1、ERBB2、PIK3CA、PIK3R1均能形成稳定靶向结合作用。实验结果表明红景天重要活性成分红景天苷具有促进自噬小体和自噬溶酶体形成的作用，并且抑制了Pl3K/AKT信号通路的活动。结论 红景天调控颗粒细胞自噬具有多成分-多靶点-多通路的作用特点，其机制与红景天苷调控PI3K/AKT信号通路有关。

Abstract:

OBJECTIVE To explore the potential targets and mechanisms of Rhodiolae Crenulatae Radix et Rhizoma in regulating autophagy in granulosa cells based on network pharmacology and experimental verification. METHODS With the help of the database, the active components, effect targets and autophagy-related targets of Rhodiolae Crenulatae Radix et Rhizoma were obtained, and a component-target network diagram and a protein-protein interaction network diagram were established. Enrichment analysis of genes and signaling pathways was conducted on key targets and the active ingredients and core targets were docked to explore the possible mechanism of Rhodiolae Crenulatae Radix et Rhizoma in regulating autophagy in granulosa cells. Western blotting, RT-PCR, transmission electron microscopy and double-labeled adenovirus were used to detect the effect of important active ingredients of Rhodiolae Crenulatae Radix et Rhizoma on autophagy in granulosa cells. RESULTS There were 7 active ingredients of Rhodiolae Crenulatae Radix et Rhizoma obtained, with 441 targets, 796 autophagy-related targets, and there were 834 biological processes, 60 cellular components, 69 molecular functions, and 150 signaling pathways obtained by enrichment analysis. Molecular docking results showed that the active components of Rhodiolae Crenulatae Radix et Rhizoma and the core targets AKT1, ERBB2, PIK3CA, PIK3R1 could form a stable target binding effect. The experimental results showed that salidroside, an important active component of Rhodiolae Crenulatae Radix et Rhizoma, could promote the formation of autophagosomes and autophagolysosomes, and inhibit the activity of the Pl3K/AKT signaling pathway. CONCLUSION The results show that the regulation of autophagy in granulosa cells by Rhodiolae Crenulatae Radix et Rhizoma has the characteristics of multi-component-multi-target-multi-pathway, and its mechanism is related to the regulation of PI3K/AKT signaling pathway by salidroside.