First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan 250014, China.
Department of reproductive medicine, Shandong University of Traditional Chinese Medicine Second Affiliated Hospital, Jinan 250001, China.
Phytomedicine. 2024 May;127:155461. doi: 10.1016/j.phymed.2024.155461. Epub 2024 Feb 22.
The active ingredients of the Chinese medical herb Paris polyphylla, P. polyphylla ethanol extract (PPE) and polyphyllin I (PPI), potentially inhibit epithelial-mesenchymal transition (EMT) in tumors. However, the roles of these ingredients in inhibiting EMT in adenomyosis (AM) remain to be explored.
The primary goal of the study was to uncover the underlying molecular processes through which PPE and PPI suppress EMT in AM, alongside assessing the safety profiles of these substances.
To assess the suppressive impact of PPE on adenomyosis-derived cells (AMDCs), we employed Transwell and wound healing assays. The polyphyllins (PPI, PPII, PPVII) contained in PPE were characterized using high-performance liquid chromatography (HPLC). Then, bioinformatics techniques were performed to pinpoint potential PPI targets that could be effective in treating AM. Immunoblotting was used to verify the key proteins and pathways identified via bioinformatics. Furthermore, we examined the efficacy of PPE and PPI in treating Institute of Cancer Research (ICR) mice with AM by observing the morphological and pathological features of the uterus and performing immunohistochemistry. In addition, we assessed safety by evaluating liver, kidney and spleen pathologic features and serum test results.
Three major polyphyllins of PPE were revealed by HPLC, and PPI had the highest concentration. In vitro experiments indicated that PPE and PPI effectively prevent AMDCs invasion and migration. Bioinformatics revealed that the primary targets E-cadherin, N-cadherin and TGFβ1, as well as the EMT biological process, were enriched in PPI-treated AM. Immunoblotting assays corroborated the hypothesis that PPE and PPI suppress the TGFβ1/Smad2/3 pathway in AMDCs to prevent EMT from progressing. Additionally, in vivo studies showed that PPE (3 mg/kg and 6 mg/kg) and PPI (3 mg/kg and 6 mg/kg), successfully suppressed the EMT process through targeting the TGFβ1/Smad2/3 signaling pathway. Besides, it was observed that lower doses of PPE (3 mg/kg) and PPI (3 mg/kg) exerted minimal effects on the liver, kidneys, and spleen.
PPE and PPI efficiently impede the development of EMT by inhibiting the TGFβ1/Smad2/3 pathway, revealing an alternative pathway for the pharmacological treatment of AM.
中药重楼的活性成分重楼乙醇提取物（PPE）和重楼苷 I（PPI），可能抑制肿瘤中的上皮-间充质转化（EMT）。然而，这些成分在抑制腺肌病（AM）中的 EMT 中的作用仍有待探索。
本研究的主要目的是揭示 PPE 和 PPI 抑制 AM 中 EMT 的潜在分子机制，并评估这些物质的安全性。
为了评估 PPE 对腺肌病来源细胞（AMDCs）的抑制作用，我们采用 Transwell 和划痕愈合实验。采用高效液相色谱法（HPLC）对 PPE 中的重楼苷（PPI、PPII、PPVII）进行了表征。然后，通过生物信息学技术确定了可能有效治疗 AM 的潜在 PPI 靶点。免疫印迹用于验证通过生物信息学确定的关键蛋白和途径。此外，我们通过观察子宫的形态和组织学特征以及进行免疫组织化学来研究 PPE 和 PPI 在治疗 AM 的 ICR 小鼠中的疗效。此外，通过评估肝、肾和脾的组织病理学特征以及血清检测结果来评估安全性。
HPLC 揭示了 PPE 的三种主要重楼苷，其中 PPI 浓度最高。体外实验表明，PPE 和 PPI 可有效阻止 AMDCs 的侵袭和迁移。生物信息学显示，主要靶点 E-钙粘蛋白、N-钙粘蛋白和 TGFβ1 以及 EMT 生物学过程在 PPI 处理的 AM 中富集。免疫印迹实验证实了 PPE 和 PPI 通过抑制 AMDCs 中的 TGFβ1/Smad2/3 通路来抑制 EMT 的假说。此外，体内研究表明，PPE（3mg/kg 和 6mg/kg）和 PPI（3mg/kg 和 6mg/kg）通过靶向 TGFβ1/Smad2/3 信号通路成功抑制 EMT 过程。此外，观察到较低剂量的 PPE（3mg/kg）和 PPI（3mg/kg）对肝脏、肾脏和脾脏的影响最小。
PPE 和 PPI 通过抑制 TGFβ1/Smad2/3 通路有效阻止 EMT 的发展，为 AM 的药物治疗提供了一种新的途径。
