大蒜素联合5-FU对腺样囊性癌ACC-M裸鼠移植瘤抑制作用的研究

发布时间：2018-04-24 07:56

本文选题：大蒜素 + 5-FU　；参考：《遵义医学院》2017年硕士论文

【摘要】：目的:探讨大蒜素联合5-FU对腺样囊性癌ACC-M裸鼠移植瘤生长的抑制作用,为其用于临床治疗腺样囊性癌提供一定的理论基础和实验依据。方法:将30只5-6周龄,体重(18±2)g的Balb/c裸鼠在SPF级动物房饲养一周。将常规培养的腺样囊性癌ACC-M细胞,用PBS缓冲液调整细胞浓度为2×107/ml,于裸鼠右前肩胛区皮下注射0.2m L接种,建立ACC-M细胞裸鼠移植瘤模型。造模2W后挑选长径(a)约5mm的裸鼠24只,随机分为实验组(大蒜素组、5-FU组和大蒜素联合5-FU组)和对照组(PBS组),每组各6只。各实验组按照体重0.01ml/g腹腔注射药物,大蒜素组、5-FU组注射大蒜素和5-FU浓度均为20mg/ml,联合组注射大蒜素20mg/ml加5-FU20mg/ml。对照组注射等量PBS液,每两天一次,共3次,末次注射6天后处死动物,完整取出瘤体组织。实验过程中观察裸鼠饮食、活动及对外界反应。从第一次给药当天至第10天,每两天测量移植瘤的体积变化,共计6次,绘制肿瘤生长曲线。HE染色观察移植瘤的病理学改变。结果:1.本实验成功建立人腺样囊性癌ACC-M裸鼠皮下移植瘤模型,在接种后2W皮下出现瘤样包块,直径约5mm,移植瘤成瘤率为100%。2.药物干预前裸鼠饮食、活动无明显改变,至处死当天对照组裸鼠饮食、活动及对外界反应较实验组明显减退。实验过程中24只裸鼠无死亡。3.移植瘤体积比较结果:随时间变化,裸鼠移植瘤体积有增大的趋势,各组移植瘤体积的变化幅度不同,对照组移植瘤体积增长迅速且明显,大蒜素组、5-FU组、大蒜素+5-FU组移植瘤增大程度低于对照组,大蒜素+5-FU组增大程度最低(p0.05);给药前各组平均瘤体积差异无统计学意义(p0.05);治疗结束后各实验组移植瘤体积明显小于对照组(p0.05),5-FU组体积明显小于大蒜素组(p0.05),大蒜素联合5-FU组与大蒜素组比较(p0.05),及与5-FU组比较(p0.05),差异均有统计学意义。4.HE染色结果显示:对照组肿瘤细胞片状分布密集,大小较一致,胞核大染色深,异常核分裂相较多见,无明显坏死区域;大蒜素组肿瘤细胞分布相对稀疏,坏死细胞占肿瘤细胞比例小于10%;5-FU组瘤体组织出现大块坏死区域,坏死细胞占肿瘤细胞比例约为30%;大蒜素联合5-FU组瘤体组织可见纤维增生,肿瘤细胞肿大变性,坏死细胞占肿瘤细胞比例约为60%。结论:1.采用裸鼠皮下接种ACC-M细胞造模是一种良好的人腺样囊性癌造模方法。2.大蒜素、5-FU单独使用以及大蒜素联合5-FU对裸鼠体内ACC-M细胞移植瘤的生长均有明显的抑制作用,大蒜素联合5-FU对移植瘤的抑制作用明显强于单独应用。
[Abstract]:Objective: to investigate the inhibitory effect of allicin combined with 5-FU on the growth of adenoid cystic carcinoma (ACC-M) xenografts in nude mice, and to provide a theoretical and experimental basis for the clinical treatment of adenoid cystic carcinoma. Methods: 30 5-6 weeks old Balb/c nude mice with weight of 18 卤2g were fed in SPF class animal room for one week. The normal cultured adenoid cystic carcinoma (ACC-M) cells were injected subcutaneously with 0.2m L into the right anterior scapular region of nude mice with PBS buffer to adjust the cell concentration to 2 脳 107 / ml. The transplanted tumor model of ACC-M cells was established in nude mice. After 2 weeks, 24 nude mice with 5mm were randomly divided into experimental group (allicin group 5-FU group and garlicin combined with 5-FU group) and control group (6 rats in each group). The concentrations of allicin and 5-FU were 20 mg / ml in allicin group and 20 mg / ml in combination group respectively according to body weight 0.01ml/g. The control group was injected with the same amount of PBS solution once every two days for 3 times. The animals were killed 6 days after the last injection and the tumor tissues were taken out completely. During the experiment, the diet, activity and reaction to the environment of nude mice were observed. From the first administration day to the 10th day, the volume changes of the transplanted tumor were measured every two days. The tumor growth curve was plotted. The pathological changes of the transplanted tumor were observed by HE staining. The result is 1: 1. In this experiment, the subcutaneous transplanted tumor model of human adenoid cystic carcinoma (ACC-M) in nude mice was successfully established. The tumor-like mass was found in the subcutaneously at 2W after inoculation, the diameter was about 5mm, and the tumor formation rate was 100.2. Before drug intervention, the diet and activity of nude mice did not change significantly, but the diet, activity and reaction to the outside world of the control group decreased significantly on the day of death. During the experiment, 24 nude mice did not die. The results of tumor volume comparison showed that the volume of transplanted tumor in nude mice increased with time. The volume of transplanted tumor in each group was different. The volume of transplanted tumor in control group increased rapidly and obviously, and that in allicin group was 5-FU group. The degree of tumor enlargement in allicin 5-FU group was lower than that in control group. The tumor volume of allicin 5-FU group was lower than that of allicin 5-FU group (P 0.05), the mean tumor volume of each group was not significantly different before administration of allicin, and the volume of transplanted tumor in each experimental group was significantly smaller than that in the control group (P 0.05) and allicin combined with allicin group (P < 0.05) after the end of the treatment, the volume of transplanted tumor in each experimental group was significantly smaller than that in the control group (P 0.05). Compared with allicin group and 5-FU group and 5-FU group, the difference was statistically significant. 4. The results of HE staining showed that the tumor cells in the control group were densely distributed. In allicin group, the distribution of tumor cells was relatively sparse, the proportion of necrotic cells in tumor cells was less than 105-FU. The percentage of necrotic cells in tumor cells was about 30%, and in allicin combined with 5-FU group, fibrous proliferation and tumor cell degeneration were observed, and the percentage of necrotic cells in tumor cells was about 60%. Conclusion 1. Subcutaneous inoculation of ACC-M cells in nude mice is a good method to model human adenoid cystic carcinoma. Allicin 5-FU alone and allicin combined with 5-FU could inhibit the growth of ACC-M cell transplantation tumor in nude mice, and the inhibitory effect of allicin combined with 5-FU was stronger than that of 5-FU alone.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.8

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