中药复方肠复康抗人大肠癌作用及机理研究
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摘要
目的 研究中药复方制剂肠复康对人大肠癌的抗癌效应，并初步探讨其作用机制。
方法 (1)建立人大肠癌HT-29裸鼠移植瘤模型。设立肠复康低、中、高剂量组和空白对照组、5Fu+CF阳性对照组。(2)肠复康灌胃给药8周后比较各组抑瘤效应的强弱。(3)采用病理形态学、免疫组化、图象分析技术定量检测并比较各组移植瘤细胞核形态、凋亡细胞数、微血管密度(MVD)以及Ki-67、iNOS、MMP-2的基因表达情况。
结果 (1)肠复康能显著抑制移植瘤生长，低、中、高剂量组瘤重均明显小于空白组(P＜0.05、P＜0.01、P＜0.001)，其中高剂量组瘤重小于5Fu+CF组(P＜0.05)。(2)肠复康可改善移植瘤细胞及细胞核形态，减少核内物质含量，高剂量组作用最为明显。肠复康各组的Ki-67计数较空白组减少，高剂量组最少并与空白组、5Fu+CF组比较有显著差异(P＜0.001、P＜0.05)。(3)肠复康低、中、高剂量组的凋亡细胞数均多于空白组(P＜0.05、P＜0.01、P＜0.001)，iNOS表达亦呈渐增趋势，其中高剂量组iNOS表达显著高于空白组(P＜0.01)。肠复康用药组凋亡细胞数与iNOS表达水平呈显著正相关(r=0.7761，P=0.0001)。(4)肠复康可降低移植瘤MVD值和MMP-2表达水平，其中高剂量组与空白组比较差异显著(P＜0.001、P＜0.01)。
结论 (1)肠复康对人大肠癌具有确切的抑瘤效果，并呈量效依赖关系。(2)肠复康具有抑制人大肠癌细胞增殖的作用，可能与改善细胞核形态，降低核内物质合成代谢有关。(3)肠复康对人大肠癌细胞具有诱导凋亡的作用，可能与促进肿瘤组织表达高水平iNOS有关。(4)肠复康具有抗人大肠癌侵袭转移的潜在作用，可能与抗肿瘤血管生成和抑制细胞外基质及基底膜降解有关。
Objective: To study the anticancer effect of Chinese Compound Recipe Changfukang against humam large intestinal cancer and preliminarily explore its mechanism.
Methods: (l)Model rats of transplantation rumor of human large intestinal cancer HT-29 were established. The low, medium and high dose groups of Changfukang ,the blank control group and 5Fu+CF positive control group were set up. (2)After 8 weeks medication of Changfukang through gastrogavage, the antitumor effect of each group was compared. (3)The transplantation tumor karyon morphology, number of apoptotic cells, micro vascular density (MVD) and the gene expression of Ki-67, iNOS, MMP-2 in each group were quantitatively detected and compared by the application of pathomorphology, immunohistochemistry and image analysis technique.
Results: (1)Changfukang could significantly inhibit the growth of transplantation tumor, and the tumor's weight in the low, medium and high dose groups was obviously lower than that in the blank group(P<0.05, P<0.01, P<0.001), and that in the 5Fu+CF group (P<0.05). (2)Changfukang could improve the cell and karyon morphology of transplantation tumor, and reduce the physical content in nucleus. This effect was most significant in the high dose group. The Ki-67 count in each Changfukang group was lower than that in the blank group, among which the high dose group had the lowest and the difference was significant compared with the blank group and 5Fu+CF group(P<0.001, P<0.05). (3)The number of apoptotic cells in Changfukang low, medium and high dose groups was separately higher than that in the blank group(P<0.05, P<0.01, P<0.001), and iNOS expression showed a upward tendency, among which the iNOS expression in high dose group was significantly , higher than that in the blank
group(P<0 01) There was a significant positive correlation between the number of apoptotic cells and the level of iNOS expression in Changfukang treated groups(r = 0.7761, P=0.0001). (4)Changfukang could lower the MVD value of transplantation tumor and the llevel of MMP-2 expresson. And this difference was significant between the high dose group and the blank group(P<0.001, P<0.01).
Conclusion: (1)Changfukang has definite antitumor effect against human large intestinal cancer and this effect is dose-dependent .(2)Changfukang has the function of inhibitingthe cell proliferation of human large intestinal cancer, which may be correlated with its effect of improving karyon morphology and reducing the physical anabolism in nucleus. (3)Changfukang has the function of inducing cell apoptosis of human large intestinal cancer, which may be correlated with its promotion of tumor tissue to express high level iNOS. (4)Changfakang has the potential function of anti-invasion and transferation of human large intestinal cancer, which may be correlated with its anti-tumor angiogenesis effect and its inhibition of the degradation of extracellular matrix and basement membrane.
方法 (1)建立人大肠癌HT-29裸鼠移植瘤模型。设立肠复康低、中、高剂量组和空白对照组、5Fu+CF阳性对照组。(2)肠复康灌胃给药8周后比较各组抑瘤效应的强弱。(3)采用病理形态学、免疫组化、图象分析技术定量检测并比较各组移植瘤细胞核形态、凋亡细胞数、微血管密度(MVD)以及Ki-67、iNOS、MMP-2的基因表达情况。
结果 (1)肠复康能显著抑制移植瘤生长，低、中、高剂量组瘤重均明显小于空白组(P＜0.05、P＜0.01、P＜0.001)，其中高剂量组瘤重小于5Fu+CF组(P＜0.05)。(2)肠复康可改善移植瘤细胞及细胞核形态，减少核内物质含量，高剂量组作用最为明显。肠复康各组的Ki-67计数较空白组减少，高剂量组最少并与空白组、5Fu+CF组比较有显著差异(P＜0.001、P＜0.05)。(3)肠复康低、中、高剂量组的凋亡细胞数均多于空白组(P＜0.05、P＜0.01、P＜0.001)，iNOS表达亦呈渐增趋势，其中高剂量组iNOS表达显著高于空白组(P＜0.01)。肠复康用药组凋亡细胞数与iNOS表达水平呈显著正相关(r=0.7761，P=0.0001)。(4)肠复康可降低移植瘤MVD值和MMP-2表达水平，其中高剂量组与空白组比较差异显著(P＜0.001、P＜0.01)。
结论 (1)肠复康对人大肠癌具有确切的抑瘤效果，并呈量效依赖关系。(2)肠复康具有抑制人大肠癌细胞增殖的作用，可能与改善细胞核形态，降低核内物质合成代谢有关。(3)肠复康对人大肠癌细胞具有诱导凋亡的作用，可能与促进肿瘤组织表达高水平iNOS有关。(4)肠复康具有抗人大肠癌侵袭转移的潜在作用，可能与抗肿瘤血管生成和抑制细胞外基质及基底膜降解有关。
Objective: To study the anticancer effect of Chinese Compound Recipe Changfukang against humam large intestinal cancer and preliminarily explore its mechanism.
Methods: (l)Model rats of transplantation rumor of human large intestinal cancer HT-29 were established. The low, medium and high dose groups of Changfukang ,the blank control group and 5Fu+CF positive control group were set up. (2)After 8 weeks medication of Changfukang through gastrogavage, the antitumor effect of each group was compared. (3)The transplantation tumor karyon morphology, number of apoptotic cells, micro vascular density (MVD) and the gene expression of Ki-67, iNOS, MMP-2 in each group were quantitatively detected and compared by the application of pathomorphology, immunohistochemistry and image analysis technique.
Results: (1)Changfukang could significantly inhibit the growth of transplantation tumor, and the tumor's weight in the low, medium and high dose groups was obviously lower than that in the blank group(P<0.05, P<0.01, P<0.001), and that in the 5Fu+CF group (P<0.05). (2)Changfukang could improve the cell and karyon morphology of transplantation tumor, and reduce the physical content in nucleus. This effect was most significant in the high dose group. The Ki-67 count in each Changfukang group was lower than that in the blank group, among which the high dose group had the lowest and the difference was significant compared with the blank group and 5Fu+CF group(P<0.001, P<0.05). (3)The number of apoptotic cells in Changfukang low, medium and high dose groups was separately higher than that in the blank group(P<0.05, P<0.01, P<0.001), and iNOS expression showed a upward tendency, among which the iNOS expression in high dose group was significantly , higher than that in the blank
group(P<0 01) There was a significant positive correlation between the number of apoptotic cells and the level of iNOS expression in Changfukang treated groups(r = 0.7761, P=0.0001). (4)Changfukang could lower the MVD value of transplantation tumor and the llevel of MMP-2 expresson. And this difference was significant between the high dose group and the blank group(P<0.001, P<0.01).
Conclusion: (1)Changfukang has definite antitumor effect against human large intestinal cancer and this effect is dose-dependent .(2)Changfukang has the function of inhibitingthe cell proliferation of human large intestinal cancer, which may be correlated with its effect of improving karyon morphology and reducing the physical anabolism in nucleus. (3)Changfukang has the function of inducing cell apoptosis of human large intestinal cancer, which may be correlated with its promotion of tumor tissue to express high level iNOS. (4)Changfakang has the potential function of anti-invasion and transferation of human large intestinal cancer, which may be correlated with its anti-tumor angiogenesis effect and its inhibition of the degradation of extracellular matrix and basement membrane.
引文
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