红花黄色素和胸腺五肽对肿瘤生长的调控作用及机制研究

【作者】 赵言群；

【导师】 牛欣；

【作者基本信息】 北京中医药大学 ， 中西医结合基础， 2006， 硕士

【摘要】 本研究旨在探讨红花黄色素及胸腺五肽对肿瘤生长的调控作用，期望能综合二者优势，为新型抗肿瘤药物的研发提供新的思路和实验参考，并在药物对肿瘤生长调控的新机理、新靶点方面做一些探讨。主要在抗肿瘤药物研究最活跃的领域新生血管生成抑制剂、细胞凋亡以及国内外肿瘤免疫方面研究的热点药物干预MHC抗原表达等方面，观察对肿瘤的影响并阐述可能的作用机制。 1 红花黄色素对小鼠H22移植性肿瘤抑制作用的实验研究 造H22荷瘤小鼠模型，观察红花黄色素SY对肿瘤的作用，实验发现：SY大、中、小剂量组小鼠一般情况均明显好于阳性对照环磷酰胺组，SY大、中剂量组小鼠亦好于模型组。各组小鼠体重增长情况由大到小依次为：SY中剂量组、SY大剂量组、SY小剂量组、模型组、环磷酰胺组。对各组小鼠肿瘤作病理切片，HE染色，发现SY中剂量组较模型组比较肿瘤组织坏死区域范围广，肿瘤细胞破碎，胞膜不完整。抑瘤率的研究发现SY中剂量组的平均瘤重为0.870g，与模型组平均瘤重1.450g相比较有显著差异，P＜0.05，中剂量组抑瘤率为31.9％，显示出明显的抑瘤活性。 结论：活血化瘀中药红花中提取的有效组分红花黄色素，对小鼠H22移植性肿瘤有明显抑制作用，其副作用较阳性对照环磷酰胺组小。 2 SY抑制小鼠H22移植性肿瘤新生血管生成的实验研究 本实验提取SY组、模型对照组小鼠肿瘤总RNA，以RT-PCR方法观察两组与肿瘤血管生成密切相关的细胞因子VEGF、KDR、bFGF、FLT、bFGFR的mRNA表达。实验发现：SY对FLT、bFGFR的作用不明显，可不同程度抑制VEGF、KDR、bFGF的mRNA表达，但抑制程度强弱不同。其中，对VEGF、KDR抑制作用较强，SY组与模型组比较有极显著的统计学差异(P＜0.01)，对bFGF也有抑制作用，与模型组比较有显著的统计学差异(P＜0.05)。 结论：红花黄色素可能通过抑制VEGF、bFGF、KDR的mRNA表达，抑制肿瘤新生血管形成，从而发挥抑制肿瘤生长的作用。 3 红花黄色素影响小鼠H22移植性肿瘤细胞生长周期的实验研究 本实验取SY组、模型对照组小鼠肿瘤组织，做成细胞悬液，以流式细胞术的方法观察各组肿瘤细胞的生长周期情况，实验发现：红花黄色素组小鼠肿瘤细胞凋亡率明显高于对照组，且细胞多停留在静止期和凋亡期。与模型组比较，SY组肿瘤细胞G0／G1期细胞数显著增多，G2／M期显著减少，S期细胞显著减少。 结论：红花黄色素可影响肿瘤细胞的生长周期，诱发细胞凋亡。其原因可能与SY抑制VEGF、bFGF的表达有关。 4 胸腺五肽对荷瘤小鼠肿瘤表面MHC-Ⅰ类分子影响的实验研究 小鼠造模方法同前，取TP-5组、模型对照组、环磷酰胺组小鼠肿瘤组织，做成更多还原

【Abstract】 The aim of this article is to study the new target and new mechanism of the regulation effect of medicine on tumor growth Especially on the two aspects, the hot spot of antitumor drug researching—the neovascularization inhibitor and the tumor immunity field ,medicine interfereing MHC antigen expressing ,we select two representative medicines which are safflor yellow and TP-5 to observe their regulating effet on tumor and study the perhaps mechanism1. The experiment of safflor yellow suppressing mouse H22 grafting tumor growth Make the tumor-bearing mice model, and observe the suppressant effect of saffloryellow to tumor The results indicate that the general state of health of the SY large midst and small dose groups are all better than the positive control cyclophosphamide group, and the SY large midst dose groups are also better than the model group The succession in turn of body weight increasing of each group from large to small are : SY midst dose group SY large dose group, SY small dose group model group , cyclophosphamide group Make pathological section and HEstaining for each group mice tumor tissue and find the zone of tumor necrosis in the SY midst dose group is more extensive than in the model group, and the tumor cells are more disruptive, and the cell membrane is more discompleted The research on tumor suppressed ratio finds the mean tumor weight is 0.870g in the SY midst dose group and the model group is 1.450g, while the difference is significant (P<0.05) And the tumor suppressed ratio of the SY midst dose group is 31.9 %, which indicates the effect of suppressing tumor is very visible Conclusion : safflor yellow can suppress mouse H22 grafting tumor growth conspicuously, and its side effect is less than the positive control cyclophosphamide group 2. The experiment of safflor yellow inhibiting the neovascularization of mouse H22 grafting tumorExtract the total RNA of each group tumor tissue, and observe the mRNA expressing of the cytokines -VEGF, KDR bFGF FLT bFGFR, which is related very closely to tumor neovascularization We find the SY effect FLT and bFGFR very little , but can inhibit the expressing of VEGF, KDF, bFGF in different degrees , and comparing to the model group , the difference is significant (P <0.05)Conclusion : SY can inhibit tumor neovascularization ,and thus suppress tumor growth, perhaps through inhibiting the mRNA expressing of VEGF KDR and bFGF3. The experiment of safflor yellow effecting the cell growth cycle of mouse H22 grafting tumorUse the tumor tissue to make cell suspension, and observe the tumor cell growth cycle更多还原