寒、热证候大鼠大肠癌启动相关机制差异分析及中药干预

发布时间：2019-03-12 11:35

【摘要】：目的:肿瘤微环境是肿瘤发生发展的一个重要影响因素,其中肿瘤相关成纤维细胞作为肿瘤微环境中最主要的组成细胞,参与肿瘤的发生发展、侵袭和转移。本研究通过比较寒、热两种证型的大鼠大肠癌启动时肿瘤相关成纤维细胞的差异,观察大鼠大肠组织中肿瘤相关成纤维细胞的三个代表性的标记物FAP、fsp-1和PDGFR β的表达情况,以寻求不同证候大鼠大肠肿瘤发生机制可能存在的差异。方法:将大鼠随机分为寒证模型组、热证模型组和空白组(雌雄各半),首先通过5周的时间,建立起寒、热证模型及空白对照模型;然后进行12周的大肠癌模型制作,并从寒证模型组和热证模型组中抽取一半大鼠,分别作为寒证中药组和热证中药组,额外给与反左金丸和左金丸进行中药反证。从第18周开始,每三周收集标本一次,观察大肠癌发病情况及FAP、fsp-1和PDGFRβ三个指标在各组中的表达情况。结果:寒、热证造模后大鼠出现较为明显的寒、热证的症状和体征,与寒热证候相关的指标ATP酶和SDH酶活性也有明显差异;在此基础上建立的大肠癌模型,通过病理检测发现大鼠大肠肿瘤发生明显,大肠癌发生早期,病理结果以寒证模型组较为严重,在后期时,热证模型组癌变率更高,而且病理分期更为严重。而对应的中药干预组的肿瘤出现时间较晚,而且肿瘤多发生于大肠以外的器官或组织。对大肠组织进行免疫组化和pcr检测,结果显示FAP、fsp-1和PDGFR β在两个模型组的表达差异主要发生在肿瘤早期,随着肿瘤进展,差异逐渐不明显;对应的中药干预组在肿瘤早期能够抑制这三个标记物的表达,而随着肿瘤进展,抑制效果越来越弱。结论:1.在寒、热证的基础上建立大肠癌模型是可行的。该模型反映出了寒、热的差异,诱导出来的大肠癌模型也具备了寒、热证候的特点,为进一步研究癌变机制提供了可用的实验动物模型。2.寒、热证大肠癌的差异主要体现在肿瘤发生早期,寒证大肠癌的发生比热证大肠癌较晚,而病理程度更严重。随着肿瘤进展,热证大肠癌的发病率和病理程度不亚于寒证大肠癌。3.左金丸和反左金丸对大肠癌的抑制作用主要体现在肿瘤早期,而且抑制效果与其药物配比存在关系。
[Abstract]:Aim: tumor microenvironment is an important factor in the development of tumor, in which tumor-associated fibroblasts, as the most important constituent cells in tumor microenvironment, participate in the occurrence, development, invasion and metastasis of tumor. In this study, we compared the difference of tumor-associated fibroblasts during the initiation of colorectal cancer in rats with cold and heat syndromes, and observed the three representative markers of tumor-associated fibroblasts (FAP,) in the large intestine tissue of rats. The expression of fsp-1 and PDGFR 尾 was studied in order to explore the possible differences in the carcinogenesis of large intestine tumors in rats with different syndromes. Methods: rats were randomly divided into three groups: cold syndrome model group, heat syndrome model group and blank group (half female and half male). Firstly, the cold, heat syndrome model and blank control model were established after 5 weeks. Then the model of colorectal cancer was made for 12 weeks, and half of the rats were taken from the cold syndrome model group and the heat syndrome model group, respectively as the cold syndrome Chinese medicine group and the heat syndrome Chinese medicine group, and the anti-syndrome of anti-Zuo Jin Pill and Zuojin Pill were given extra. The specimens were collected every three weeks from the 18th week to observe the incidence of colorectal cancer and the expression of FAP,fsp-1 and PDGFR 尾 in each group. Results: the rats with cold and heat syndrome showed obvious cold, symptoms and signs of heat syndrome, and the activities of ATP enzyme and SDH enzyme, which were related to cold-heat syndrome, were also significantly different. The results showed that there was no significant difference between cold-heat syndrome and cold-heat syndrome. The model of colorectal cancer was established on the basis of pathological examination. It was found that the occurrence of colorectal cancer in rats was obvious in the early stage of colorectal cancer, and the pathological results were more serious in the cold syndrome model group, and in the later stage, the canceration rate in the heat syndrome model group was higher than that in the heat syndrome model group. And pathological stage is more serious. The corresponding traditional Chinese medicine intervention group appeared later, and most of the tumors occurred in organs or tissues outside of the large intestine. The results of immunohistochemistry and pcr showed that the difference of expression of FAP,fsp-1 and PDGFR 尾 between the two model groups mainly occurred in the early stage of the tumor, but the difference was not obvious with the progression of the tumor. The corresponding Chinese medicine intervention group could inhibit the expression of these three markers in the early stage of tumor, but with the progression of tumor, the inhibition effect became weaker and weaker. Conclusions: 1. It is feasible to establish colorectal cancer model on the basis of cold and heat syndrome. This model reflects the difference between cold and heat, and the induced colorectal cancer model also has the characteristics of cold and heat syndrome, which provides an available experimental animal model for further study of the mechanism of carcinogenesis. 2. The difference between cold and heat syndrome of colorectal cancer is mainly reflected in the early stage of tumorigenesis. The occurrence of cold syndrome of colorectal cancer is later than that of heat syndrome, but the degree of pathology is more serious. With the progression of tumor, the incidence and pathological degree of colorectal cancer in heat syndrome is no less than that in cold syndrome. 3. The inhibitory effects of Zuo Jin Pill and Anti-Zuo Jin Pill on colorectal cancer were mainly reflected in the early stage of tumor, and the inhibitory effect was related to the proportion of drugs.
【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R273

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