【作者】 张若楠；

【导师】 王三虎；

【作者基本信息】 第四军医大学 ， 中西医结合， 2004， 硕士

【摘要】 目的：虽然现代医学对于恶性肿瘤的防治已取得了迅猛的发展，但疗效并不尽如人意。而中医药或中西医结合治疗肿瘤积累了丰富的经验，在肿瘤防治中逐渐显示出独特优势和巨大潜力，能在一定程度上弥补现代医学治疗肿瘤的不足。独活寄生汤是一首治疗风寒湿痹而兼有体虚的通用方剂，具有祛风湿，益肝肾，止痹痛，补气血的功效，王三虎教授在多年的肿瘤临床中认为癌症骨转移疼痛的基本病机与独活寄生汤所治符合，应用此方标本间治，缓解了不少癌症骨转移患者的疼痛。本实验在此基础上，就独活寄生汤的抑瘤作用及相关机制进行了初步探讨。 方法：1)独活寄生汤煎剂的制备：采用正交设计方法，以有效成分总多糖的含量及出膏率为指标，苯酚—硫酸法测定总多糖含量，对提取工艺进行了优化。2)急性毒性实验：给实验小鼠灌服不同浓度的独活寄生汤，测定小鼠灌胃最大耐受量。3)实体瘤抑瘤实验：将S₁₈₀细胞悬液接种于昆明小鼠皮下，建立S₁₈₀实体瘤肿瘤模型。将荷S₁₈₀实体瘤的昆明小鼠分为5组：对照组（生理盐水组），5—Fu组，中药大剂量组（高浓度），中药中剂量组（中浓度），中药小剂量组（低浓度）。于接种S₁₈₀肉瘤细胞的第6d给药，10d后，处死小鼠，取瘤称重，计算瘤重抑制率。同时剖离瘤组织，病理切片观察。4)独活寄生汤对荷瘤小鼠免疫功能影响的实验：抑瘤实验的各组动物活杀后无菌取胸 第四军医大学硕士学位论文里里.巴里里巴，里里里巴里里里里腺、脾脏称湿重，计算脾和胸腺脏体系数。运用改良的MTT法测定荷瘤小鼠腹腔巨噬细胞活性，测定ConA诱导的荷瘤小鼠T淋巴细胞增殖及荷瘤小鼠NK细胞活性。采用ConA诱导T细胞生物活性微量测定法测定荷瘤小鼠IL一2活性。 结果:l)独活寄生汤煎剂的制备:最佳工艺条件为煎煮3次，加水量分别为生药质量的10倍、8倍和8倍，煮沸时间分别为60，30和30 min。2)急性毒性实验:实验小鼠灌胃给药的最大耐受量倍数为62倍。3)实体瘤抑瘤实验:统计各组实体瘤的平均重量，表明各治疗组药物对荷瘤小鼠的肿瘤生长均有一定的抑制作用，它们与生理盐水组相比，组间差异具有统计学意义，5一Fu组与独活寄生汤组相比，组间差异具有统计学意义。病理切片结果显示，与生理盐水组对照比较，各用药组显示肿瘤组织不同程度的变性坏死，以独活寄生汤中剂量组和5一Fu组为著。3)独活寄生汤对荷瘤小鼠免疫功能影响的实验:①对荷瘤小鼠免疫器官指数的影响:独活寄生汤组荷瘤小鼠的脾指数明显高于生理盐水组。独活寄生汤组荷瘤小鼠的胸腺指数高于生理盐水组，其中中、小剂量组明显高于生理盐水组，大剂量组也高于生理盐水组，但无统计学差异;5一Fu治疗组脾指数、胸腺指数均低于生理盐水组，也低于独活寄生汤组。②对荷瘤小鼠腹腔巨噬细胞、T淋巴细胞、NK细胞的影响:独活寄生汤组3个剂量均促进荷瘤小鼠腹腔M中细胞的活性，与生理盐水、5一Fu组相比有显著差异。独活寄生汤组与生理盐水组相比显著促进荷瘤小鼠脾细胞的增值，与5一Fu相比也有显著性差异。独活寄生汤组与生理盐水组相比，其中、小2个剂量可明显增强荷瘤小鼠NK细胞活性，与5一Fu组相比也有显著性差异。5一Fu治疗组荷瘤小鼠腹腔M中细胞的活性，NK细胞的活性及脾细胞的增值均低于生理盐水值，其中荷瘤小鼠腹腔M中细胞的活性，NK细胞的活性与生理盐水组相比有显著差异。③对荷瘤小鼠IL一2的影响:独活寄生汤组可显著提高荷瘤小鼠淋巴细胞对IL一2的反应性，与生理盐水组、5一Fu组相比有统计学差异，5一Fu组与生理 第四军医大学硕士学位论文目.里，..里里里巴粤里里巴.巴盐水组相比有抑制荷瘤小鼠IL一2生成的作用趋势，但两者无统计学差异。 结论:l)以所述工艺条件提取制备，即确保有效成分，也保证了一定的出膏率，为临床合理煎煮独活寄生汤，进一步提高疗效提供了可靠的实验依据。2)独活寄生汤可以有效抑制小鼠5180实体瘤的生长，并在一定范围内存在着量效关系，提示该方可能是一种有效的抗癌复方。3)独活寄生汤对荷瘤小鼠的细胞免疫功能、非特异性免疫功能及与肿瘤免疫相关的细胞因子均有一定的促进作用，说明该方可作用于免疫系统的多个环节，这可能抑制小鼠移植性肿瘤生长的重要机制之一。更多还原

【Abstract】 Object: Although modern medical treatment of tumor has been developing rapidly,there are still some dissatisfactions at present. However, TCM and Integrated Traditional Western Medicine treatment of tumor have been gradually coming to ripe, and tremendous potentiality in prevention and treatment of tumor in very long history. They have showed distinctive preponderance and tremendous potentiality in prevention and treatment of tumor, and are able to compensate some misplays of modern medical treatment of tumor. Duhuojisheng decoction is an universal prescription used to treat wind-cold-damp obstruction with weakness. It has efficacies of dispelling wind and dampness.relieving pain,nourishing liver and kidney,invigorating qi and blood. Prof. Wang Sanhu thanks that the essential pathomechanism between ostealgia of cancer transfer and therapy of Duhuojisheng decoction may match. It’s confirmed that Duhuojisheng decoction may lighten pathomechanism on cancer transfer through his perennial clinical practice. Therefore, the present experiment is intended to study the effects and mechanisms of Duhuojisheng decoction in inhibiting carcinoma.Methods: 1)Preparation of Duhuojisheng decoction,the optimal extracting technology was selected using the total polysaccharides and therate of paste as the standards by orthogonal design. Phenol-sulfuric acid method was used to determine the content of total polysaccharides. 2) Acute toxicity test:Gave experiment mice different concentration of Duhuojisheng decoction,then determined the maximum tolerant dose. 3)In the inhibitory experiment of the solid carcinoma: The model of mice bearing carcinoma was established by inoculating sarcoma-180 cells into the subcutaneous tissue or the abdominal cavity of the Kunming mice.The Kunming mice bearing sarcoma-180 cells were divided into five groups:high dose group of Duhuojisheng decoction;medium dose group of Duhuojisheng decoction;low dose group of Duhuojisheng decoction;5-Fu group;control group(NS group).The various drugs and natural saline were given into mice stomach at the 6th day after inoculating sarcoma-180 cells.Then ten days later,execute the mice,cut the tumor,weigh it and calculate the inhibition ratios of the tumor weight. Meanwhile,tumor tissue dissected and separated, pathological section observed in microscope. 4) Experiment on effect of Duhuojisheng decoction on immune function in inhibiting mice carcinoma:Mice bearing tumor were executed,mice thymus and spleen were cut and weighed ,spleen index and thymus index were calculated. Modified MTT assay were used to determine M activity,T-lymphocyte’s generation and NK activity. Activity of IL-2 was determined by biologic activity method.Results:1 ) Preparation of Duhuojisheng decoction,the optimal conditions were as follows: The decoction was thrice, the water quantity was 10 times, 8 times and 8 times as much as the drug quantity, the boiling time was 60, 30 and 30 min in each time. 2) Acute toxicity test, maxima tolerance does of Duhuojisheng decoction for mice was 62 times of that for human body. 3) In the inhibitory experiment of the solid carcinoma, the distinction between experiment group and control grouphad a significantly meaning by analyzing average weight of the each group solid carcinoma,which indicated that the drugs of experiment group could inhibit sarcoma-180. The distinction between Duhuojisheng decoction and 5-Fu group had a significantly meaning Pathological section result indicated that tumor tissue got degeneration or necrosis in some extent and medium dose group of Duhuojisheng decoction and 5-Fu group showed more significant than NS group. 4) Experiment on effect of Duhuojisheng decoction on immune function in inhibiting mice carcinoma.(1)Effect of Duhuojisheng decoction on immunoapparatus in tumor-bearing mice. Spleen index of Duhuojisheng decoction group was enhanced. Compared with NS group, they had significant difference.Thymus index was enhanced in the medium and low dosages of Duhuojisheng decoction group compared with NS group. Spleen index an更多还原