摘要
目的:探讨左金丸及其主要单体成分在体内体外对大肠癌发生发展的干预作用.方法:培养人大肠癌细胞株HT29,选择处于对数生长期的细胞,分别用不同浓度的盐酸小檗碱(26.25、52.5、105、210、420μmol/L)、吴茱萸碱(5、10、15、20、25、30μmol/L)处理HT29细胞24、48、72、96h,筛选最适作用浓度与时间,在这一浓度与时间作用下,用活细胞计数法检测左金丸的主要单体成分小檗碱和吴茱萸碱对HT29细胞增殖的抑制作用,TRAP法检测HT29细胞端粒酶活性,取HT29细胞,用生理盐水制成单细胞悬液,调整细胞浓度为5×105/0.2mL,注射于裸鼠左侧胁腹部皮下,制作HT29细胞荷瘤裸鼠肿瘤模型.随机将裸鼠分为4组:对照组、左金丸组、AZT组和协同组,观察左金丸和叠氮胸苷(AZT)对瘤体生长及端粒酶活性的影响.利用致癌剂1,2-二甲基酰肼(DMH,25mg/kg)诱导大鼠大肠癌模型,同样随机将大鼠分为4组,观察左金丸和AZT对大鼠大肠癌发生发展的干预作用.结果:在离体实验中,处理24-72h,105μmol/L盐酸小檗碱和15μmol/L吴茱萸碱对HT29细胞增殖的抑制作用呈成正向的线性量效关系,吴茱萸碱(7.5、15、30μmol/L)作用后的抑制率分别为39.3%±2.13%,52.8%±5.34%和64.1%±7.19%,盐酸小檗碱(52.5、105、210μmol/L)作用后的抑制率分别为44.1%±3.97%,55.9%±4.12%和65.3%±6.94%.105μmol/L小檗碱和15μmol/L吴茱萸碱处理HT29细胞72h,小檗碱和吴茱萸碱可以有效抑制HT29细胞端粒酶的活性.在荷瘤裸鼠实验中,左金丸和AZT对瘤体大小无明显影响,对端粒酶活性有微弱影响.第11周,对照组、左金丸组、AZT组和协同组大鼠肿瘤发生率分别为20%、0%,10%和10%,左金丸作用优于AZT,但34wk各组间无明显差异.结论:离体实验中,左金丸的主要成分可以有效抑制HT29细胞的增殖和端粒酶的活性;裸鼠移植瘤实验中,观察不到左金丸的药效;1,2-DMH诱导大肠癌实验模型中,左金丸可以有效阻碍早期癌症的发生和发展.
AIM： To investigate the intervention effect of Zuojin Pill and its major constituents against colorectal carcinoma in vitro and in vivo. METHODS： Cultured HT29 cells in the logarithmic growth phase were treated with berberine （26.25, 52.5, 105, 210 and 420 μmol/L, respectively） and evodiamine （5, 10, 15, 20, 25 and 30μmol/L, respectively） for 24, 48, 72 and 96 h, respectively, to determine the optimal concentration and duration of treatment. Under optimal conditions, the inhibitory effects of berberine and evodiamine on the proliferation of HT29 ceils were examined by a viable cell count method, and the telomerase activity of the treated cells was determined by the TRAP method. The single cell suspension of HT29 cells was then prepared and subcutaneously injected into the left abdominal region of nude mice at a concentration of 5×105 cells/0.2 mL to produce tumorbearing nude mice. These mice were randomly divided into four groups, namely, control group （injection with saline）, Zuojin Pill group, AZT （Azidothymidine） group and Zuojin Pill plus AZT group. The effects of Zuojin Pill and AZT on tumor growth and telomerase activity were observed. Additionally, a rat model of colorectal carcinoma was induced with 1, 2-dimethyl hydrazine （DMH）. The rats were also randomly divided into four groups, treated in the same manner as described above, and killed at weeks 11, 21 and 34, respectively, to observe the effects of Zuojin Pill and AZT against the progression of rat colorectal carcinoma. RESULTS： Evodiamine and berberine dosedependently inhibited the proliferation of HT29 cells in vitro. The suppression ratios achieved using evodiamine （7.5, 15 and 30μmol/L, respectively） were 39.3% ± 2.13%, 52.8% ± 5.34% and 64.1% ± 7.19%, respectively, while those achieved using berberine （52.5, 105, 210 μmol/L） were 44.1% ± 3.97%, 55.9% ± 4.12% and 65.3% ± 6.94%, respectively. Moreover, berberine and evodiamine could effectively inhibit the telomerase activity of HT29 cells. In tumor-bearing nude mice, Zuojin Pill and AZT had no significant influence on tumor size but had weak effect on telomerase activity. In DMH-induced rat colon cancer model, the tumor incidences observed at week 11 in the control group, Zuojin Pill group, AZT group and Zuojin Pill plus group were 20%, 0%, 10% and 10%, respectively. In contrast, the tumor incidences at week 34 showed no significant difference among different groups. CONCLUSION： The major constituents of Zuojin Pill can suppress the proliferation and telomerase activity of HT29 ceils in vitro. In DMH- produced rat model, Zuojin Pill can effectively repress the development and progression of early-stage carcinoma, but has no obvious effect against late-stage carcinoma.
出处
《世界华人消化杂志》
CAS
北大核心
2009年第19期1936-1941,共6页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30400602
No.30672687~~
