【作者】 周清安；

【导师】 孙志广；

【作者基本信息】 南京中医药大学 ， 中西医结合临床， 2009， 博士

【摘要】 一、研究目的、内容和意义1研究整理文献,寻找其病因病机和防治方药的脉络和规律,分析古今用药规律及各自特点,探求有效的防治途径和方法。2体外实验中,从Bcl-2、Bax和Caspase-3的表达,对皂苷调控LoVo细胞凋亡作一探讨。3探讨猫爪草皂苷对移植性结肠癌小鼠模型(CT-26)的抗肿瘤作用及其诱导凋亡的相关机理。二、研究结果1文献研究1.1病因病机古代医家认为主要有饮食因素、起居不节、感受外邪、先天因素、情志因素等,且多数赞同本病是多种因素共同作用的结果。现代医家发展了大肠癌的病因病机理论,认为主要包括气滞学说、血瘀学说、热毒学说、湿聚学说、正气虚弱学说。1.2治法方药古代主要有补益虚损、理气活血、清热解毒、收涩止血、通下等法。今人则以清热解毒为主,温下法使用较少。古今用药各有侧重。补益虚损是古今医家在论治时均认识到的重要方法;今人治疗大肠癌,清热药占绝对优势,且温里药与古人用药相比有明显下降。2实验研究2.1猫爪草皂苷对人结肠癌细胞LoVo增殖呈明显抑制作用,且随药物浓度的提高和作用时间的延长而增强,500μg/mL72h其对LoVo抑制率为81.52%。2.2猫爪草皂苷作用人结肠癌细胞LoVo后,在荧光显微镜观察到细胞凋亡的特征形态改变,在流式图上各浓度组均见有亚二倍体峰的出现。24h出现DNA ladder。2.3猫爪草皂苷作用24h后,LoVo细胞Bax蛋白水平明显增高,而Bcl-2出现抑制,且呈浓度依赖性,皂苷组Bcl-2/Bax比例下降。Caspase-3蛋白水平随浓度明显升高。2.4猫爪草皂苷对小鼠移植性肿瘤CT-26有显著抑制作用。在本实验剂量范围内出现明显量效关系,其中最大剂量平均抑瘤率分别为39.97%,同时对荷瘤小鼠CT-26免疫器官胸腺、脾脏指数均有增加,NK细胞活性上升。Bax、Caspase-3蛋白水平明显升高,Bcl-2/Bax比值显著下降。三、研究结论1防治思路在于防治并举,防重于治;标本兼顾,祛邪寓扶正之中;以通为要,辨寒温各施温下、寒下;宜重视温下法的合理运用。2寒毒积滞是大肠癌类病证的主要病机之一。勿轻温热,重寒凉。温下法是治疗大肠癌的有效思路及方法。3猫爪草皂苷具有显著抑制人结肠癌LoVo细胞生长、诱导其凋亡的作用,具有时间、剂量依赖关系。猫爪草皂苷阻滞细胞周期,抑制肿瘤的生长。4猫爪草皂苷可降低人结肠癌LoVo细胞Bcl-2表达,增加Bax、Caspase-3表达。猫爪草皂苷可能通过压力诱导的途径,激活凋亡执行器Caspase-3诱导凋亡。5猫爪草皂苷抑制小鼠CT-26肿瘤的生长和诱导小鼠结肠癌细胞凋亡,其机理可能是下调Bcl-2的表达,上调Bax的表达。更多还原

【Abstract】 The purpose, content and result lie in the following. Ancient doctors holded colorectal cancer pathogen mainly in diet, accommodation, Waixie, congenital, emotional factors, while using Buyi, Wenli, Liqi drugs, Qingre; promoting blood circulation, Shouse, diarrhea, hemostatic. Modern in Qi stagnation, blood stasis, hot, wet, Zhengqi weak, therefore Qingre; Buyi; promoting blood circulation, hemostatic, Liqi.Saponins inhibited proliferation of LoVo, and paralleled to concentration and time, 500μg/mL72h on LoVo inhibition rate of 81.52%. The apoptosis of the characteristics was seen. Each drug group could be seen the emergence of hypodiploid peak by flow cytometry. 24h, DNA ladder emergenceed, and Bax rised, while Bcl-2 inhibied significantly with a dose-dependent manner, saponin group Bcl-2/Bax decreased. Caspase-3 was significantly increased with the concentration.Saponins on the mouse tumor CT-26 was significant inhibition with a dose-effect relationship, the maximum dose’s average tumor inhibition rate at 39.97%, the tumor-bearing mice thymus, spleen index increased, NK cell activity rised. Bax and Caspase-3 were significantly increased. Bcl-2/Bax ratio decreased significantly.Conclusion can be deduce. Prevention is more important; Biaoben into account; Key Tong, identified the Han and Wen; Attention to good use of Wenxia. Han Ji Zhi is one of the main pathogenesis of colorectal cancer patients.Saponins significantly inhibited the growth of LoVo cells, induced by Apoptosis with time, dose-dependent relationship, arrested cell cycle, and inhibited tumor growth. Saponins can reduce the LoVo cells Bcl-2 expression while increased Bax, Caspase-3 protein expression.That saponins inhibited CT-26 mouse tumor growth and induceed apoptosis of colon cancer in mice is related to the reduction of Bcl-2 expression, increase the expression of Bax.更多还原