肠泰抑制人结直肠癌裸鼠移植瘤生长的实验研究

发布时间：2018-04-12 22:24

本文选题：肠泰 + 结直肠癌　；参考：《南京中医药大学》2016年硕士论文

【摘要】：目的：肠泰是江苏省中西医结合医院治疗结直肠癌的经验方。本文通过3PLC对肠泰进行质量控制;建立异位移植瘤动物模型,研究肠泰对人结直肠癌HT-29细胞裸鼠移植瘤的抑制作用并探索其机制。方法：1)采用HPLC方法对肠泰进行质量控制。2)采用异位移植瘤动物模型,研究肠泰对人结直肠癌HT-29细胞裸鼠体内异位移植瘤生长的影响,检测经不同浓度的肠泰作用后裸鼠体内肿瘤的体积大小及生长情况；采用一步法TUNEL检测荷瘤小鼠不同浓度肠泰作用下肿瘤组织凋亡水平。3)提取人结直肠癌HT-29细胞荷瘤小鼠肿瘤蛋白,采用Western-Blot技术,检测肿瘤细胞中Cleaved Caspase-9、Cleaved Caspase-3、PARP、Cleaved PARP、 Bcl-XL、Bax、LC3 Ⅰ、LC3 Ⅱ的蛋白表达水平,评估肿瘤细胞凋亡、自噬情况。结果：1)采用外标一点法,分析出肠泰中人参皂苷Rg1、苦参碱、人参皂苷Rb1、黄芪甲苷、阿魏酸和毛蕊异黄酮苷的含量分别为0.273、0.457、0.492、0.791、0.008、0.166mg/g生药量。2)肠泰能够显著抑制裸鼠异位移植瘤的生长,且随药物浓度增高抑瘤率明显增高；TUNEL检测随药物浓度增高,移植瘤肿瘤细胞凋亡程度增高。3)肠泰能够激活Cleaved Caspase-9、Cleaved Caspase-3和PARP,增加促凋亡因子Bax的表达,抑制抗凋亡因子Bcl-XL的表达；同时可增加自噬相关蛋白LC3Ⅱ的表达,并呈浓度依赖性。结论：1)可采用HPLC法对肠泰进行质量控制,质控成分为人参皂苷Rg1、苦参碱、人参皂苷Rb1、黄芪甲苷、阿魏酸和毛蕊异黄酮苷,其含量分别为0.273、0.457、0.492、0.791、0.008、0.166 mg/g生药量；2)肠泰能抑制人结直肠癌HT-29细胞裸鼠移植瘤的生长,促进移植瘤肿瘤细胞凋亡；3)肠泰通过诱导HT_29移植瘤细胞发生自噬和凋亡来抑制肿瘤细胞的增殖。
[Abstract]:Objective: Changtai is the experience of hospital treatment of colorectal cancer combined with Chinese and Western medicine in Jiangsu province. This thesis uses 3PLC to control the quality of Changtai; heterotopic transplantation tumor animal model, inhibition of Changtai on human colorectal cancer HT-29 cells in nude mice and explore its mechanism. Methods: 1) by using HPLC method quality control of.2 of Changtai) using xenograft animal model of Changtai of node influence the growth of colorectal cancer cells HT-29 in nude mice xenograft tumor, detection of the size and growth of different concentrations of Changtai after in vivo tumor; by one-step TUNEL.3 levels of apoptosis in tumor tissue was detected in tumor bearing mice under the action of different concentration of Changtai) extraction of human colon tumor bearing mice colorectal cancer cells HT-29 protein, using Western-Blot technology, detection of Cleaved Caspase-9 in tumor cells, Cleaved, Caspase-3, PARP, C Leaved PARP, Bcl-XL, Bax, LC3 I, the expression level of LC3 II protein, apoptosis, evaluation of autophagy in tumor cells. Results: 1) by using one point external standard method, analysis of intestinal Thailand matrine, ginsenoside Rg1, ginsenoside Rb1, astragaloside and ferulic acid and calycosin glycoside were 0.273,0.457,0.492,0.791,0.008,0.166mg/g crude drug.2) Changtai can significantly inhibit the growth of xenograft tumor in nude mice, and the inhibition rate increased with the drug concentration increased significantly; TUNEL detection with the drug concentration increased, xenograft tumor cell apoptosis increased.3) Changtai can activate Cleaved Caspase-9, Cleaved Caspase-3 and PARP, increase the expression of Pro apoptotic factor Bax the expression of anti apoptosis factor Bcl-XL; at the same time can increase the expression of autophagy related protein LC3 II, which was dose-dependent. Conclusion: 1) can be used to control the quality of Changtai by HPLC method, quality control Composition of matrine, ginsenoside Rg1, ginsenoside Rb1, astragaloside and ferulic acid and campanulin, their contents were 0.273,0.457,0.492,0.791,0.008,0.166 mg/g crude drug; 2) inhibition of intestinal Tienam HT-29 human colorectal cancer cell growth in nude mice, promote the apoptosis of tumor cell transplantation; 3) Changtai to inhibit tumor cell proliferation by inducing tumor cell transplantation HT_29 autophagy and apoptosis.

【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R273

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