【作者】 梁颖；

【导师】 王亚贤；

【作者基本信息】 黑龙江中医药大学 ， 中西医结合基础， 2012， 博士

【摘要】 目的：研究红花多糖(SPS)对荷S180肉瘤BALB/c小鼠肿瘤转移相关基因CD44、MMP-9、TIMP-1、AMFmRNA及nm23-H1蛋白含量的影响,观察SPS对血管生成及抗肿瘤的作用,进一步探讨SPS抑制肿瘤转移作用及其机制。方法：1.Real Time PCR法检测BALB/c小鼠肿瘤组织CD44、MMP-9. TIMP-1、AMFmRNA的表达情况2.Western blot法检测肿瘤组织中nm23-H1蛋白表达情况。3.鸡胚绒毛尿囊膜实验观察血管生成数量及面积的变化。4.免疫组化法检测小鼠肿瘤组织VEGF蛋白及MVD的变化。5.ELISA法检测荷瘤小鼠血清中IL-12、IL-10及TNF-α的含量结果：1.Real Time PCR检测发现,SPS作用于荷瘤小鼠后,肿瘤组织中CD44、MMP-9、AMF mRNA的表达下降,TIMP-1mRNA的表达升高2.Western blot检测发现,经SPS作用后小鼠肿瘤组织中nm23-H1蛋白含量上升。3.SPS中剂量组干预后鸡胚绒毛尿囊膜上血管生成数目和面积较模型对照组减少(P<0.05),SPS高剂量组较模型对照组显著减少(P<0.01)。4.免疫组化法检测发现,SPS中剂量组小鼠肿瘤组织中VEGF阳性表达率明显下降(与模型组比较P<0.01),MVD降低极其明显(P<0.001)；SPS高剂量组小鼠肿瘤组织中VEGF阳性表达率及MVD降低极其明显(P<0.001)。5.ELISA法检测发现,与模型对照组相比较,SPS中、高剂量组小鼠血清中IL-12和TNF-α含量升高极其明显(P<0.001)；IL-10含量下降极其显著(P<0.001)。结论：1.SPS能够抑制小鼠肿瘤组织CD44、MMP-9、AMF mRNA的表达,促进TIMP-1mRNA的表达,在肿瘤转移的黏附、降解、运动环节上抑制肿瘤的转移。2.SPS能够促进小鼠肿瘤组织nm23-H1的表达,抑制肿瘤的转移3.SPS具有抑制鸡胚绒毛尿囊膜上血管生成的作用。4.SPS能够通过抑制VEGF的表达抑制荷瘤小鼠肿瘤组织血管的生成,降低MVD,使肿瘤生长及转移受到抑制。5.SPS能够通过提高IL-12的表达,降低IL-10的表达调整Th1/Th2漂移,并促进TNF-α表达。更多还原

【Abstract】 Objective:To study the SPS which has an effect on BALB/c mice bearing S180tumor metastasis related genes CD44、MMP-9、 TIMP-1、AMF mRNA and nm23-H1protein, to observe the SPS on angiogenesis and anti-tumor effect, to explore the SPS refrain tumor metastasis and its mechanism furthermore.Methods:1. Detect the CD44and MMP-9, TIMP-1, AMF mRNA expression of BALB/c mice tumor tissue by Real Time PCR assay2. Detect the expression of nm23-H1protein of tumor tissue by Western blot assay.3. Chicken embryos chorioallantoic membrane assay to observe angiogenesis number and size of the change.4. Immunohistochemical assay to detect the mice tumor tissue VEGF protein and MVD changes.5. ELISA assay to detect serum IL-12, IL-10and TNF-α content.Results:1. Real Time PCR detection,the SPS role in tumor-bearing mice, tumor tissue of CD44、MMP-9、AMF mRNA expression was decreased and TIMP-1mRNA expression was increased.2. Western blot analysis found that the nm23-H1protein content increased in the tumor tissue of mice with the reaction of the SPS.3. SPS mid-dose group after the intervention of the SPS in the chicken embryos chorioallantoic membrane angiogenesis, the number and size compared with the model control group decreased (P<0.05),SPS high-dose group compared with the model control group was significantly reduced (P<0.01)4. Immunohistochemical methods detected the SPS mid-dose groups VEGF expression in tumor tissue was significantly decreased (compared with model group P<0.01), MVD reduction is extremely obvious (P<0.001); VEGF positive expression rates and MVD reduction in the SPS high-dose group of mice tumor tissue are extremely clear (P<0.001)5. ELISA examination found that compared with the model group, the mid-dose, high dose of the serum IL-12and TNF-α levels increased very significantly (P<0.001), IL-10decreased very significantly (P<0.001)Conclusion:1. SPS can inhibit tumor tissue of mice CD44, MMP-9, the AMF mRNA expression, promote TIMP-1mRNA expression. SPS can inhibit the adhesion, degradation, and motion of tumor metastasis.2. SPS can restrain tumor metastasis by promoting the expression of nm23-H1in tumor tissue of mice3. SPS can inhibit the generation of the chicken embryos chorioallantoic membrane vascular.4. By inhibiting VEGF expression and reducing MVD, SPS can restrain the growth of blood vessels in the tumor tissue, make tumor growth and transfer under control.5. SPS can increase the expression of IL-12, reducing the expression of IL-10to adjust the Th1/Th2shift, and to promote TNF-α expression.更多还原