Jiao Juying, Cheng Chien-Shan, Xu Panling, Yang Peiwen, Ruan Linjie, Chen Zhen
Department of Integrative Oncology, Fudan University Shanghai Cancer Center, Shanghai, China.
Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China.
Front Oncol. 2022 Jul 25;12:947238. doi: 10.3389/fonc.2022.947238. eCollection 2022.
Damp-heat syndrome is one of the most important syndrome types in the traditional Chinese medicine (TCM) syndrome differentiation and treatment system, as well as the core pathogenesis of pancreatic cancer (PC) which remains a challenge to medical researchers due to its insidious onset and poor prognosis. Great attention has been given to the impact of damp-heat syndrome on tumorigenesis and progression, but less attention has been given to damp-heat modeling per se. Studying PC in a proper damp-heat syndrome animal model can recapitulate the actual pathological process and contribute to treatment strategy improvement.
Here, an optimized damp-heat syndrome mouse model was established based on our prior experience. The Fibonacci method was applied to determine the maximum tolerated dosage of alcohol for mice. Damp-heat syndrome modeling with the old and new methods was performed in parallel of comparative study about general appearance, food intake, water consumption and survival. Major organs, including the liver, kidneys, lungs, pancreas, spleen, intestines and testes, were collected for histological evaluation. Complete blood counts and biochemical tests were conducted to characterize changes in blood circulation. PC cells were subcutaneously inoculated into mice with damp-heat syndrome to explore the impact of damp-heat syndrome on PC growth. Hematoxylin-eosin staining, Masson staining and immunohistochemistry were performed for pathological evaluation. A chemokine microarray was applied to screen the cytokines mediating the proliferation-promoting effects of damp-heat syndrome, and quantitative polymerase chain reaction and Western blotting were conducted for results validation.
The new modeling method has the advantages of mouse-friendly features, easily accessible materials, simple operation, and good stability. More importantly, a set of systematic indicators was proposed for model evaluation. The new modeling method verified the pancreatic tumor-promoting role of damp-heat syndrome. Damp-heat syndrome induced the proliferation of cancer-associated fibroblasts and promoted desmoplasia. In addition, circulating and tumor-located chemokine levels were altered by damp-heat syndrome, characterized by tumor promotion and immune suppression.
This study established a stable and reproducible murine model of damp-heat syndrome in TCM with systematic evaluation methods. Cancer associated fibroblast-mediated desmoplasia and chemokine production contribute to the tumor-promoting effect of damp-heat syndrome on PC.
湿热证是中医辨证论治体系中最重要的证型之一，也是胰腺癌（PC）的核心发病机制。胰腺癌起病隐匿，预后较差，给医学研究人员带来了挑战。湿热证对肿瘤发生和进展的影响已受到高度关注，但湿热证模型本身却较少受到关注。在合适的湿热证动物模型中研究胰腺癌，可以重现实际的病理过程，有助于改进治疗策略。
在此，基于我们之前的经验建立了一种优化的湿热证小鼠模型。应用斐波那契方法确定小鼠酒精的最大耐受剂量。采用新旧方法并行进行湿热证建模，并对一般外观、食物摄入量、饮水量和生存率进行比较研究。收集包括肝脏、肾脏、肺、胰腺、脾脏、肠道和睾丸在内的主要器官进行组织学评估。进行全血细胞计数和生化检测以表征血液循环的变化。将胰腺癌细胞皮下接种到患有湿热证的小鼠体内，以探讨湿热证对胰腺癌生长的影响。进行苏木精-伊红染色、Masson染色和免疫组织化学以进行病理评估。应用趋化因子微阵列筛选介导湿热证促增殖作用的细胞因子，并进行定量聚合酶链反应和蛋白质免疫印迹以验证结果。
新的建模方法具有对小鼠友好、材料易获取、操作简单和稳定性好的优点。更重要的是，提出了一套系统的指标用于模型评估。新的建模方法验证了湿热证对胰腺肿瘤的促进作用。湿热证诱导癌相关成纤维细胞增殖并促进纤维组织形成。此外，湿热证改变了循环和肿瘤部位的趋化因子水平，其特征是促进肿瘤和免疫抑制。
本研究建立了一种具有系统评估方法的稳定且可重复的中医湿热证小鼠模型。癌相关成纤维细胞介导的纤维组织形成和趋化因子产生有助于湿热证对胰腺癌的促肿瘤作用。
