| 铜绿假单胞菌制剂对小鼠膀胱癌和脾细胞作用的机制研究 | |
| Alternative Title | Effect of Pseudomonas Aeruginosa on murine splenic cells and development of bladder cancer model in mice |
| 李涛 | |
| Subtype | 硕士 |
| Thesis Advisor | 王志平 |
| 2014-05-28 | |
| Degree Grantor | 兰州大学 |
| Place of Conferral | 兰州 |
| Degree Name | 硕士 |
| Keyword | 膀胱肿瘤 原位模型 铜绿假单胞菌制剂 免疫治疗 |
| Abstract | 目的：研究铜绿假单胞菌制剂（PA-MSHA）抗膀胱癌效应和脾脏细胞增殖效果的影响。 方法：通过硝酸银、盐酸、胰酶和乙醇预处理C57BL/6小鼠膀胱粘膜，摘取小鼠膀胱组织，HE染色观察粘膜病理变化；戊二醛固定，观察微观结构变化；甲苯胺蓝染色，观察膀胱粘膜固有层的肥大细胞数目变化；过碘酸-希夫染色，观察膀胱粘膜GAG层变化。应用前四组预处理膀胱，构建原位肿瘤模型计算各组成瘤率。建立小鼠肿瘤皮下模型，监测肿瘤体积变化；检测CD4+和CD8+ ；计数VEGF评分和微血管密度（MVD）。制备脾细胞悬液，计算增值效率。 结果：胰酶和酒精作用1小时后，膀胱粘膜局部上皮伞状细胞脱落；酸碱和硝酸银组粘膜下层暴露较多。24小时后胰酶和乙醇处理组粘膜完整性恢复较好；酸碱和硝酸银处理组膀胱粘膜薄厚不一。PA-MSHA作用组肿瘤体积与对照组比较明显缩小，效果呈现浓度梯度变化；高浓度PA-MSHA作用组小鼠生存时间最长；VEGF和MVD在PA-MSHA作用组显著降低；术后21天和28天，PA-MSHA作用组小鼠血清CD4+、CD8+水平显著提高；脾细胞增值实验中， PA-MSHA作用组OD值较对照组显著增高，且呈现时间和剂量依赖性。 结论：硝酸银和酸碱可作为构建鼠膀胱肿瘤原位模型的首选预处理因素。PA-MSHA能够抑制肿瘤的生长，提高小鼠的生存率和免疫力，发挥抗肿瘤作用。 |
| Other Abstract | Objective: To explore the optimization for establishment of mouse models of orthotopic bladder tumors, to evaluate the effects of pseudomonas aeruginosa (PA-MSHA) on bladder cancer in mouse subcutaneous tumor model and on the proliferation of splenic lymphocytes, to explore the mechanism involved in, and to provide new treatment strategies for bladder cancer. Methods: After 1 or 24 hours the pretreatment of murine bladder mucosa with silver nitrate, HCl, trypsin and ethanol, the mice were killed to obtain bladder tissue samples. Pathological changes were observed using hematoxylin and eosin staining. The microenvironment modification was assessed by electron microscopy. Mast cells were counted with toluidine blue staining. The glycosaminoglycan (GAG) layer was observed by periodic acid-Schiff staining. Mice were intravesically pretreated with the above four reagents, respectively, and the rate of tumor formation was determined. Subcutaneous bladder tumor model was established, and divided into four groups: normal saline treatment group, low PA-MSHA dose treatment group, medium PA-MSHA dose treatment group, and high PA-MSHA dose treatment group. PA-MSHA or normal saline were injected around the tumors. Three mice were sacrificed in each group at 14, 21 and 28 after implantation, serum was collected from each mice at 3000×g for 5 minutes, then CD4+ or CD8+ were detected through flow cytometry analysis. Tumor tissue, and major organs were obtained to observe pathological changes, VEGF and MVD were also assessed. The spleens were removed aseptically from C57BL/6 mice, then spleen cell suspensions were treated with low PA-MSHA dose, medium PA-MSHA dose, high PA-MSHA dose and RPMI-1640 culture medium as control for 24, 48, 72 h. At the end point, the optical density (OD) values of the samples were measured on a microplate reader. Results: In the model mice, a part of umbrella cells were sloughed off, the submucosal layer was exposed and incontinuous in some areas at one hour after pretreatment with trypsin and ethanol. Mucosal damages were more serious in the HCl and silver nitrate groups. However, there was no significant difference in the inflammatory cell infiltration between the experiment and control groups. Mild edema and congestion in the mucosa, tightly arranged epithelial cells, and continuous mucosa were observed in the mice after trypsin and ethanol pretreatment for 24 hours. By contrast, the thickness of mucosa was uneven ... |
| URL | 查看原文 |
| Language | 中文 |
| Document Type | 学位论文 |
| Identifier | https://ir.lzu.edu.cn/handle/262010/222626 |
| Collection | 学院待认领 |
| Affiliation | 临床医学院 |
| Recommended Citation GB/T 7714 | 李涛. 铜绿假单胞菌制剂对小鼠膀胱癌和脾细胞作用的机制研究[D]. 兰州. 兰州大学,2014. |
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