【作者】 王程；

【导师】 杨运高；

【作者基本信息】 第一军医大学 ， 中西医结合基础， 2005， 硕士

【摘要】 目的: 在国家自然科学基金课题的资助下,选用桂枝茯苓丸、丹参汤活血化瘀2个不同作用力度和层次的方剂,通过观察其对红细胞免疫功能正常及缺陷两种状态下大肠癌肝转移小鼠模型的作用,初步阐明活血化瘀方药对大肠癌脾移植肝转移的影响是抑制还是促进,并探讨其作用与红细胞免疫的关系。 方法: 1.活血化瘀方药的制备选择活血化瘀的桂枝茯苓丸(桂枝9g、赤芍9g、丹皮9g、桃仁9g、茯苓9g)和养血活血的丹参汤(丹参30g)。实验时换算成小鼠每日等效剂量。 2.模型制作及分组 断奶1周的BALB/c纯系健康小鼠96只,雌雄不拘,随机分为Ⅰ、Ⅱ组2大组,Ⅰ组给予正常饲料,Ⅱ组给予低硒饲料以构建红细胞免疫缺陷模型,共饲养10周。10周后每大组(即Ⅰ、Ⅱ组)各48只小鼠组内又随机分4小组,每组12只小鼠。Ⅰ组均为红细胞免疫功能正常小鼠:①正常对照组,②病理对照A组(仅接种瘤细胞),③桂枝茯苓丸A组及④丹参汤A组均接种瘤细胞+活血化瘀法处理;Ⅱ组为红细胞免疫功能缺陷组:⑤红更多还原

【Abstract】 Object:Supported by the National natural science foundation, we choose classical traditional Chinese medicine (TCM) prescriptions of promoting blood circulation and removing blood stasis, namely Guizhifuling Pills and Danshen Tang, observe their effects on the metastatic behavior of malignant tumor, explore the possible mechanisms of such effects and their interaction with erythrocyte immunity.Method:1. Preparation of TCM prescription Guizhifuling Pills (Guizhi 9g, Chishao 9g, Danpi 9g, Shaoyao 9g, Fuling 9g) and Danshen Tang (Danshen 30g) were chosen. It will switched into the equivalent dosage while feeding the mice.2. Replication of the mouse model and grouping of the miceLiver metastases were replicated in BALB/c mice by intrasplentic injection ofcolonic adenocarcinoma cells(CT-26) with 1 X 106 cells in 0.1 ml solution. Erythrocyte immunodeficiency mice model were replicated by feeding the low-selenium forage for 10 weeks.96 BALB/c mice were randomly divided into 2 groups, namely group I and II which were divided into 4 small groups respectively, each of the small group contain 12 mice. Group I were the mice with normal erythrocyte immunity: (D Normal control group, (2) Pathological control A group (receive intrasplenic injection of CT-26 cells), (3) Guizhifuling Pill A group and ? Danshen Tang A group (receive both of the intrasplenic injection of CT-26 cells and TCM prescription). Group II were mice with erythrocyte immunodeficiency: ? RBC-hypoimmunity group, (6) Pathological control B group( erythrocyte immunodeficiency + intrasplenic injection of CT-26 cells), ? Guizhifuling pill B group and ? Danshen Tang B (erythrocyte immunodeficiency + intrasplenic injection of CT-26 cells + treatment of TCM prescription).3. The indicators of the observation3.1 The liver weight and number of the metastic tumor were surveyed.3.2 The detection of RBC immune adherence function (RIAF)The indicators of RIAF included RBC C3b receptor rosette(R-C3bR), RBC immune complex receptor rosette(R-ICR) and RBC immune adherence tumor cell receptor rosette(RITCR). A RBC adhered to more than 2 microzyme were defined as positive and the RBC rosettes were counted.3.3 The expression of human telomerase reverse transcriptase(hTERT), P53, C-erbB-2 and Bcl-2 were detected by SABC immunohistochemical(IH) method. Brown was defined as positive expression in the IH method.Result:1. The hepatic metastasis models of colonic adenocarcinoma with different RIF was successfully replicated. We used CT-26 murine colon adenocarcinoma cells syngeneic to BALB/c mice, and it is characterized by high and stable repetition and metastasis incidence, and similar metastasis pathway to patients. The method for mice model replication is feasible with stability, reliability and repeatability and can meet the demand of related researches.2. Tumor metastasis in the group mice with erythrocyte immunodeficiency were severer with significant difference (P<0.05) compared with the normal group mice inoculated with tumor cells. After all group animals were administrated with TCM prescriptions, the findings indicate that Guizhifuling Pills and Danshen tang can inhibit tumor metastasis in normal group animals inoculated with tumor cells. However, effects of the two prescriptions don’t display remarkable difference compare with the pathological control B group (P >0.05), and there was remarkable difference between erythrocyte immunodeficiency mice and the normal mice both inoculated with tumor cells and treated by the two prescriptions (P <0.05).3. The expression of hTERT, P53, C-erbB-2 and Bcl-2 in the models receiving treatment with Guizhifuling Pills differed significantly from those of pathological control A group (P <0.05), whereas between Danshen tang and the model group, only the expression of hTERT showed significant difference.Conclusion:1. Being erythrocyte immunodeficiency, the liver metastasis of colorectal carcinoma was observed obviously. The blood-activating and stasis-eliminating herbs could inhibit carcinoma metastasis exactly. In group I , the tumor inhibitive effect ofGuizhifuling Pills is better than Dansken tang(P <0.05). However, in group II, there is no significant difference for the tumor inhibitive effect between Guizhifuling Pills and Danshen tang.2. Guizhifuling Pills can reduce the expressions of hTERT, p53, c-erbB-2 and Bcl-2 in the mice models, while Danshen tang can only reduce the expression of hTERT without similar effects on the other 3 oncogenes. Such inhibitory effects of the prescriptions on hTERT and the oncogenes might explain their actions to inhibit malignant tumor metastasis.3. Mechanism of inhibiting tumor metastasis: ? related to the function of erythrocyte immunity, (2) controlling the expression of telomerase, (3) controlling the expression of P53,C-erbB-2,Bcl-2.4. The inhibiting effects of the TCM of blood circulation promotion by removal of blood stasis on tumor metastasis depend on erythrocyte immune function in great extent.更多还原