Huang Yu, Jiang Yuanyuan, Ji Hui, Gao Yu, Xiao Long, Zha Wei, Zhou Jinhua, Huang Haiwei
Department of Obstetrics and Gynecology, Zhangjiagang Hospital Affiliated to Soochow University, Zhangjiagang, Jiangsu, 215006, China.
Translational Medical Innovation Center, Zhangjiagang TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Zhangjiagang, Jiangsu, 215006, China.
Int Immunopharmacol. 2024 Dec 25;143(Pt 3):113527. doi: 10.1016/j.intimp.2024.113527. Epub 2024 Nov 2.
Chikusetsusaponin IVa (CHS IVa) as a natural extract from the Panax japonicus (T.Nees) C.A.Mey (P. japonicus), can regulate the immune responses, such as anti-inflammation, which have been applied in treating various diseases. It is still unclear, nevertheless, whether the CHS IVa can target-able treat endometriosis (EMs) and what the possible mechanism would be.
This work aims to investigate the possible mechanism and the impact of CHS IVa on EMs.
The EMs models were established in mice by autologous transplantation or chemicals (lipopolysaccharide and adenosine triphosphate), inducing the pyroptotic endometrial stromal cells. Then the CHS IVa was used to treat the EMs mice. The therapeutic impact of CHS IVa was assessed by hematoxylin-eosin staining, immunofluorescent staining, western blot (WB), and enzyme-linked immunosorbent assay (ELISA).
The results of immunofluorescence and WB indicated that pyroptosis indicators, including Gasdermin-D (GSDMD), Caspase-1, NOD-like receptor thermal protein domain associated protein 3 (NLRP3), and interleukin (IL)-1β, were substantially expressed in the ectopic endometrial lesions of EMs mice. The ELISA results showed that the abdominal cavity of EMs mice had higher concentrations of IL-1β, IL-6, and TNF-α than the non-EMs animals (control group). As shown in the molecule docking experiments, CHS IVa exhibited high binding affinity with GSDMD, IL-1β, Caspase-1, and NLRP3. Moreover, after treatment with CHS IVa, the expression levels of GSDMD, IL-1β, Caspase-1, and NLRP3 decreased in the EMs mice. Meanwhile, the expression level of pain-related proteins, such as pro-nerve growth factor (pro-NGF) and transient receptor potential vanilloid-1 (TRPV1), was inhibited via the treatment of CHS IVa. According to the antibody chip analysis, the insulin-like growth factor 1 receptor/phosphatidylinositide 3-kinases (IGF1R/PI3K) signal pathway was essential to the CHS IVa's treatment of EMs. Finally, according to the WB experiments, after the treatment with CHS-Ⅳa, the expression of IGF1R, PI3K, and related phosphorylated proteins increased compared to the mice in lipopolysaccharide + adenosine triphosphate (LPS + ATP) groups.
CHS IVa can activate the IGF1R/PI3K signal pathway, inhibit the pyroptosis of endometrial stromal cells, and relieve the inflammation and EMs.
竹节人参皂苷IVa（CHS IVa）作为从竹节人参（T.Nees）C.A.Mey（竹节人参）中提取的天然提取物，可调节免疫反应，如抗炎作用，已被应用于治疗各种疾病。然而，CHS IVa是否能够靶向治疗子宫内膜异位症（EMs）以及可能的机制仍不清楚。
本研究旨在探讨CHS IVa对EMs的可能作用机制及其影响。
通过自体移植或化学物质（脂多糖和三磷酸腺苷）在小鼠体内建立EMs模型，诱导子宫内膜基质细胞发生焦亡。然后用CHS IVa治疗EMs小鼠。通过苏木精-伊红染色、免疫荧光染色、蛋白质免疫印迹（WB）和酶联免疫吸附测定（ELISA）评估CHS IVa的治疗效果。
免疫荧光和WB结果表明，焦亡指标，包括Gasdermin-D（GSDMD）、半胱天冬酶-1、NOD样受体热蛋白结构域相关蛋白3（NLRP3）和白细胞介素（IL）-1β，在EMs小鼠的异位子宫内膜病变中大量表达。ELISA结果显示，EMs小鼠腹腔内IL-1β、IL-6和肿瘤坏死因子-α的浓度高于非EMs动物（对照组）。分子对接实验表明，CHS IVa与GSDMD、IL-1β、半胱天冬酶-1和NLRP3具有较高的结合亲和力。此外，用CHS IVa治疗后，EMs小鼠中GSDMD、IL-1β、半胱天冬酶-1和NLRP3的表达水平降低。同时，通过CHS IVa治疗，疼痛相关蛋白，如前神经生长因子（pro-NGF）和瞬时受体电位香草酸受体1（TRPV1）的表达水平受到抑制。根据抗体芯片分析，胰岛素样生长因子1受体/磷脂酰肌醇3激酶（IGF1R/PI3K）信号通路对CHS IVa治疗EMs至关重要。最后，根据WB实验，与脂多糖+三磷酸腺苷（LPS+ATP）组小鼠相比，用CHS-Ⅳa治疗后，IGF1R、PI3K和相关磷酸化蛋白的表达增加。
CHS IVa可激活IGF1R/PI3K信号通路，抑制子宫内膜基质细胞焦亡，减轻炎症反应和EMs。
