Department of Obstetrics and Gynecology, Xuzhou Central Hospital, The Xuzhou Clinical College of Xuzhou Medical University, Xuzhou, Jiangsu, China.
Department of Obstetrics and Gynecology, Graduate School of Bengbu Medical University, Bengbu, China.
BMC Complement Med Ther. 2024 Jun 11;24(1):227. doi: 10.1186/s12906-024-04509-y.
Endometrial cancer (EC) is an oestrogen-dependent tumour, the occurrence of which is closely related to an imbalance of oestrogen homeostasis. Our previous studies explored the effects of Resveratrol(Res) on oestrogen metabolism. However, systematic research on the exact mechanism of action of Res is still lacking. Based on network pharmacology, molecular docking and animal experiments, the effects and molecular mechanisms of Res on endometrial cancer were investigated.
The target of Res was obtained from the high-throughput experiment and reference-guided database of TCM (HERB) and the Encyclopedia of Traditional Chinese Medicine (ETCM) databases, and the target of endometrial cancer was obtained by using the Genecards database. Venny map was used to obtain the intersection target of Res in the treatment of endometrial cancer, and the protein interaction network of the intersection target was constructed by importing the data into the STRING database. Then, the drug-disease-target interaction network was constructed based on Cytoscape 3.9.1 software. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed for intersection targets using the OmicShare cloud platform. Res and core targets were analysed by molecular docking. EC model mice induced by MNNG were randomly divided into the control group, Res group, MNNG group, MNNG + Res group, and MNNG + Res + MAPK/ERKi group. The protein levels of ERK and p-ERK in the mouse uterus were detected by Western blot. The levels of E1, E2, E3, 16-epiE3, 17-epiE3, 2-MeOE1, 4-MeOE1, 2-MeOE2, 4-MeOE2, 3-MeOE1, 2-OHE1, 4-OHE1, 2-OHE2, 4-OHE2, and 16α-OHE1 in the serum and endometrial tissue of mice were measured by LC‒MS/MS.
A total of 174 intersection targets of Res anti-endometrial cancer were obtained. The signalling pathways analysed by KEGG enrichment included the AGE-RAGE signalling pathway in diabetic complications, the PI3K-Akt signalling pathway and the MAPK signalling pathway. The top 10 core targets were MAPK3, JUN, TP53, CASP3, TNF, IL1B, AKT1, FOS, VEGFA and INS. Molecular docking showed that in addition to TNF, other targets had good affinity for Res, and the binding activity with MAPK3 was stable. Western blot results showed that Res increased the phosphorylation level of ERK and that MAPK/ERKi decreased ERK activation. In the LC-MS/MS analysis, the levels of 2-MeOE1, 2-MeOE2 and 4-MeOE1 in serum and uterine tissue showed a significantly decreasing trend in the MNNG group, while that of 4-OHE2 was increased (P < 0.05). The concentrations of 4-MeOE1 in serum and 2-MeOE1 and 2-MeOE2 in the endometrial tissue of mice were significantly increased after Res treatment, and those of 4-OHE2 in the serum and uterus of mice were significantly decreased (P < 0.05). Meanwhile, in the MAPK/ERKi intervention group, the effect of Res on the reversal of oestrogen homeostasis imbalance was obviously weakened.
Res has multiple targets and multiple approaches in the treatment of endometrial cancer. In this study, it was found that Res regulates oestrogen metabolism by activating the MAPK/ERK pathway. This finding provides a new perspective for subsequent research on the treatment of endometrial cancer.
子宫内膜癌（EC）是一种雌激素依赖性肿瘤，其发生与雌激素稳态失衡密切相关。我们之前的研究探讨了白藜芦醇（Res）对雌激素代谢的影响。然而，Res 的确切作用机制仍缺乏系统研究。基于网络药理学、分子对接和动物实验，研究了 Res 对子宫内膜癌的作用及其分子机制。
从中药高内涵筛选和参考指导数据库（HERB）和中药综合数据库（ETCM）中获得 Res 的靶标，从基因卡片数据库中获得子宫内膜癌的靶标。使用 Venny 图谱获取 Res 治疗子宫内膜癌的交集靶标，并将数据导入 STRING 数据库构建蛋白相互作用网络。然后，基于 Cytoscape 3.9.1 软件构建药物-疾病-靶标相互作用网络。使用 OmicShare 云平台对交集靶标进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析。对 Res 和核心靶标进行分子对接分析。采用 MNNG 诱导 EC 模型小鼠，随机分为对照组、Res 组、MNNG 组、MNNG+Res 组和 MNNG+Res+MAPK/ERKi 组。采用 Western blot 检测小鼠子宫中 ERK 和 p-ERK 的蛋白水平。采用 LC-MS/MS 法检测小鼠血清和子宫内膜组织中 E1、E2、E3、16-epiE3、17-epiE3、2-MeOE1、4-MeOE1、2-MeOE2、4-MeOE2、3-MeOE1、2-OHE1、4-OHE1、2-OHE2、4-OHE2 和 16α-OHE1 的水平。
获得了 174 个 Res 抗子宫内膜癌的交集靶标。KEGG 富集分析的信号通路包括糖尿病并发症中的 AGE-RAGE 信号通路、PI3K-Akt 信号通路和 MAPK 信号通路。前 10 个核心靶标是 MAPK3、JUN、TP53、CASP3、TNF、IL1B、AKT1、FOS、VEGFA 和 INS。分子对接显示，除 TNF 外，其他靶标与 Res 具有良好的亲和力，与 MAPK3 的结合活性稳定。Western blot 结果表明，Res 增加了 ERK 的磷酸化水平，而 MAPK/ERKi 降低了 ERK 的激活。在 LC-MS/MS 分析中，MNNG 组血清和子宫组织中 2-MeOE1、2-MeOE2 和 4-MeOE1 的水平呈明显下降趋势，而 4-OHE2 的水平升高（P<0.05）。Res 处理后，小鼠血清中 4-MeOE1 和子宫内膜组织中 2-MeOE1 和 2-MeOE2 的浓度明显升高，而小鼠血清和子宫中 4-OHE2 的浓度明显降低（P<0.05）。同时，在 MAPK/ERKi 干预组中，Res 对雌激素稳态失衡的逆转作用明显减弱。
Res 在治疗子宫内膜癌方面具有多个靶标和多种作用机制。本研究发现 Res 通过激活 MAPK/ERK 通路调节雌激素代谢。这一发现为后续子宫内膜癌治疗的研究提供了新视角。
