Protective effect of paeoniflorin on H2O2 induced Schwann cells injury based on network pharmacology and experimental validation
Protective effect of paeoniflorin on H2O2 induced Schwann cells injury based on network pharmacology and experimental validation
复制标题
基于网络药理学和实验验证的芍药苷对H_2O_2诱导雪旺细胞损伤的保护作用
DOI：
10.1016/s1875-5364(21)60010-9
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发表时间：
2021-02-25
影响因子：
4.6
通讯作者：
Zhao Guo-Ping
中科院分区：
文献类型：
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作者：
Zhang Di;Yang Bing;Zhao Guo-Ping
本研究旨在探讨芍药苷对过氧化氢损伤的保护作用。首先，在Pubchem中检索PF的“SMILES”，并进一步在Swiss Target Prediction数据库中进行反向分子对接，以获得潜在的靶标。从GeenCards数据库中获取损伤相关分子，通过韦恩图筛选PF治疗损伤的预测靶点。对于机制分析，蛋白质-蛋白质相互作用由String构造，并且KEGG分析在Webgestalt中进行。CCK 8法检测细胞活力和细胞毒活性。同时将实验细胞分为对照组、模型组、（200 μ mol.L-1 H2O2），SB 203580 10 μ mol.L-1（200 μ mol.L-1 H2O2 + SB 203580 10 μ mol.L-1），PF 50 μ mol.L-1（200 μ mol·L ~（-1）H_2O_2 + PF 50 μ mol·L ~（-1））和PF 100 μ mol·L ~（-1）（200 μ mol·L ~（-1）H_2O_2 + PF 100 μ mol·L ~（-1））组。检测细胞内活性氧（ROS）、Hoechst 33258染色、细胞凋亡、Bcl-2、Caspase-3、Cleaved-caspase-3、Cleaved-caspase-7、TRPA 1、TRPV 1的表达以及p38 MAPK磷酸化水平。有96个潜在的目标，可能与PF损伤治疗。然后，我们从前10条KEGG通路中选择了“炎症介质调节TRP通道”通路进行实验验证。实验验证，H2 O2可中度降低细胞活力（P < 0.05），100 μ mol·L-1 PF可显著提高细胞活力（P < 0.05）。PF可抑制H_2O_2诱导的雪旺细胞活性氧的产生，其机制与细胞内活性氧荧光强度的差异有关。在Hoechst 33258染色中，PF逆转了H2 O2处理后染色质浓缩和凋亡细胞核。流式细胞仪检测结果显示PF能显著抑制H2 O2诱导的细胞凋亡（P < 0.05）。PF预处理可显著降低H2 O2处理后Caspase 3、Cleaved-caspase 3、Cleaved-caspase 7、TRPA 1、TRPV 1的表达水平，降低H2 O2处理后p38 MAPK磷酸化表达水平（P < 0.05），升高Bcl-2、Bcl-xl的表达水平（P < 0.05）。PF可抑制H2 O2诱导的雪旺细胞损伤和凋亡，其机制与抑制p38 MAPK磷酸化有关。
This study was to investigate the protective effect of paeoniflorin (PF) on hydrogen peroxide-induced injury. Firstly, "SMILES" of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets. Injury-related molecules were obtained from GeenCards database, and the predicted targets of PF for injury treatment were selected by Wayne diagram. For mechanism analysis, the protein-protein interactions were constructed by String, and the KEGG analysis was conducted in Webgestalt. Then, cell viability and cytotoxicity assay were established by CCK8 assay. Also, the experimental cells were allocated to control, model (200 mu mol.L-1 H2O2), SB203580 10 mu mol.L-1 (200 mu mol.L-1 H2O2 + SB203580 10 mu mol.L-1), PF 50 mu mol.L-1 (200 mu mol.L-1 H2O2 + PF 50 mu mol.L-1), and PF 100 mu mol.L-1 (200 mu mol.L-1 H2O2 + PF 100 mu mol.L-1) groups. We measured the intracellular ROS, Hoechst 33258 staining, cell apoptosis, the levels of Bcl-xl, Bcl-2, Caspase-3, Cleaved-caspase3, Cleaved-caspase7, TRPA1, TRPV1, and the phosphorylation expression of p38MAPK. There are 96 potential targets that may be associated with PF for injury treatment. Then, we chose the "Inflammatory mediator regulation of TRP channels" pathway for the experimental verification from the first 10 KEGG pathway. In experimental verification, H2O2 decreased the cell viability moderately (P < 0.05), and 100 mu mol.L-1 PF increased the cell viability significantly (P < 0.05). Depending on the difference of intracellular ROS fluorescence intensity, PF inhibited H2O2-induced reactive oxygen species production in Schwann cells. In Hoechst 33258 staining, PF reversed the condensed chromatin and apoptotic nuclei following H2O2 treatment. Moreover, Flow cytometry results showed that PF could substantially inhibit H2O2 induced apoptosis (P < 0.05). Pretreatment with PF obviously reduced the levels of Caspase3, Cleaved-caspase3, Cleaved-caspase7, TRPA1, TRPV1, and the phosphorylation expression of p38MAPK after H2O2 treatment (P < 0.05), increased the levels of Bcl-2, and Bcl-xl (P < 0.05). PF inhibited Schwann cell injury and apoptosis induced by hydrogen peroxide, which mechanism was linked to the inhibition of phosphorylation of p38MAPK.