Peng Mengfan, Zhang Xiaofang, Yang Lei, Liu Baosong
Faculty of Medicine, Huanghuai University, Zhumadian, Henan, China.
Department of Pharmacy, Zhumadian Traditional Chinese Medicine Hospital, Zhumadian, Henan, China.
Front Oncol. 2025 Jun 18;15:1585057. doi: 10.3389/fonc.2025.1585057. eCollection 2025.
Explore whether Oridonin (Ori) improves esophageal cancer by interfering in the TLR4/NF-κB/NLRP3 inflammasome.
An esophageal mouse model was induced by 4-nitroquinoline N-oxide (4-NQO) for 16 weeks. Starting from the 17th week of modeling, the mice were randomly divided into three groups: the model group (M), the high dose group of Ori (Ori -H) and the low dose group of Ori (Ori -L). The weight, diet, and water intake of the mice were recorded at the end of the experiment. H&E staining was used for esophageal tissue to evaluate pathological status. The tumor markers, inflammatory factor, mRNA and protein expression of TLR4/NF-κB/NLRP3 inflammasome related indicators in serum and esophageal tissue was determined by ELISA, qPCR, and western blot (WB) respectively. The blood cell analyzer was used for measuring the proportion of various blood cells.
Ori can increase the weight, the intake amount of food and water of mice (<0.05, <0.01). In parallel, Ori can alleviate pathological changes of esophageal tissue, decrease the levels of inflammatory factor tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), and interleukin-6 (IL-6) in serum (<0.01), and down-regulate granulocyte (Gran), Gran-to- Lymphocyte (Lymph) ratio (NLR), monocyte (Mon)-to-lymph ratio (MLR), and platelets-to-Lymph ratio (PLR) in the peripheral blood, while increasing Lymph, red blood cell (RBC), hemoglobin (HGB) (<0.05, <0.01). Moreover, the protein expression of toll-like receptor 4 (TLR4), phosphorylated nuclear factor-κB (p-NF-κB), IL-1β, NOD-like receptor hot protein domain related protein 3 (NLRP3), aspartate specific cysteine protease-1 (Caspase-1), apoptosis-associated speck-like protein (ASC), N-cadherin, and p-GSK3β was significantly inhibited by Ori (<0.05, <0.01), and the mRNA expression of proliferating cell nuclear antigen (PCNA), Ki67, and B-cell lymphoma-2 (Bcl-2) was significantly inhibited, while Bax mRNA was increased by Ori (<0.05, <0.01).
This study provides evidence indicating that Ori may inhibit inflammatory response by inhibiting TLR4/NF-κB/NLRP3 inflammasome activation, ultimately exert anti esophageal cancer effects.
探讨冬凌草甲素（Ori）是否通过干扰Toll样受体4（TLR4）/核因子κB（NF-κB）/NOD样受体热蛋白结构域相关蛋白3（NLRP3）炎性小体来改善食管癌。
用4-硝基喹啉-N-氧化物（4-NQO）诱导建立食管小鼠模型，持续16周。从建模第17周开始，将小鼠随机分为三组：模型组（M）、冬凌草甲素高剂量组（Ori-H）和冬凌草甲素低剂量组（Ori-L）。实验结束时记录小鼠体重、饮食和饮水量。采用苏木精-伊红（H&E）染色评估食管组织病理状态。分别用酶联免疫吸附测定（ELISA）、实时荧光定量聚合酶链反应（qPCR）和蛋白质免疫印迹法（WB）检测血清和食管组织中TLR4/NF-κB/NLRP3炎性小体相关指标的肿瘤标志物、炎性因子、mRNA和蛋白表达。使用血细胞分析仪检测各类血细胞比例。
冬凌草甲素可增加小鼠体重、食物摄入量和饮水量（<0.05，<0.01）。同时，冬凌草甲素可减轻食管组织病理变化，降低血清中炎性因子肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）、环氧化酶-2（COX-2）和白细胞介素-6（IL-6）水平（<0.01），下调外周血中粒细胞（Gran）、粒细胞与淋巴细胞比值（NLR）、单核细胞与淋巴细胞比值（MLR）以及血小板与淋巴细胞比值（PLR），同时增加淋巴细胞（Lymph）、红细胞（RBC）、血红蛋白（HGB）含量（<0.05，<0.01）。此外，冬凌草甲素显著抑制Toll样受体4（TLR4）、磷酸化核因子κB（p-NF-κB）、IL-1β、NOD样受体热蛋白结构域相关蛋白3（NLRP3）、天冬氨酸特异性半胱氨酸蛋白酶-1（Caspase-1）、凋亡相关斑点样蛋白（ASC）、N-钙黏蛋白和磷酸化糖原合成酶激酶3β（p-GSK3β）的蛋白表达（<0.05，<0.01），并显著抑制增殖细胞核抗原（PCNA）、Ki67和B细胞淋巴瘤-2（Bcl-2）的mRNA表达，而冬凌草甲素使Bax mRNA表达增加（<0.05，<0.01）。
本研究提供的证据表明，冬凌草甲素可能通过抑制TLR4/NF-κB/NLRP3炎性小体激活来抑制炎症反应，最终发挥抗食管癌作用。
