【作者】 千昌石；

【导师】 金星林；

【作者基本信息】 延边大学 ， 肝胆外科， 2009， 硕士

【摘要】 目的:探讨虎眼万年青总皂苷在肝损伤硬化,肝癌发生、发展中的防治作用及其影响。方法:二乙基亚硝胺诱发大鼠肝癌变,虎眼万年青总皂苷干预,运用常规HE染色法观察肝组织病理形态改变;采用比色分析法检测丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)的活性;运用免疫组化方法观察并分析肝组织标本中增殖细胞核抗原(Ki67)、血管内皮生长因子受体-1(Flt-1)的表达。结果:(1)12周肝硬化阶段模型组以出现增生结节,嗜碱性变,透明变,水变性为主;14周后模型组出现大量的“灶中灶”现象,18周模型组全部出现癌变。而虎眼万年青总皂苷预防组12周时减少嗜碱性细胞结节以出现少量假小叶、纤细纤维间隔、灶状变性为主。14周预防组假小叶、不规则纤维间隔、肝细胞灶状水变性、脂肪变性为主。14周治疗组以部分处肝硬化、变性轻度为特点。18周肝癌治疗组出现肝硬化、增生结节、灶状变性,同时癌周出现包裹性的囊肿。(2)在肝损伤硬化、肝癌的发生发展阶段MDA含量逐渐增高;SOD在12周模型组显著增高,与趋于正常组水平的模型14周、模型18周组均有统计学差异。SOD活力在虎眼万年青作用的12周预防组低于12周模型组,且有统计学差异。而在14周、18周虎眼万年青总皂苷作用组与相应模型组间,无显著差异。GSH-Px活力在模型组12周、14周、18周逐渐下降,相互间有显著性差异。虎眼万年青总皂苷作用组明显提高GSH-Px活力,并与相应的模型组间均有显著性差异。(3)肝癌变过程中以12周模型组Ki67LI表达最高,14周及18周模型组Ki67 LI逐渐降低,与12周模型组比较有统计学意义。虎眼万年青总皂苷作用的12周预防组Ki67 LI较低且与相应12周模型组有统计学意义。诱癌14周模型组Flt-1 LI表达最高,12周及18周模型组Flt-1 LI相对低下,与14周模型组比较有统计学意义。虎眼万年青总皂苷作用的14周预防组、治疗组Flt-1 LI降低,且与模型组有统计学意义。结论:(1)虎眼万年青总皂苷以癌周出现囊肿阻断癌灶血液供应起到抑制肝癌发展,诱导肝癌细胞凋亡,起到治疗肝癌的作用。(2)虎眼万年青总皂苷具有降低MDA的生成作用,尤以肝损伤硬化阶段明显;虎眼万年青总皂苷干预SOD活力水平发生在肝损伤硬化阶段;虎眼万年青总皂苷在肝癌促进、发生发展阶段中能够增加GSH-Px的活力,提高抗氧化系统功能,降低对毒物的敏感性。(3)肝癌变过程中以肝损伤硬化阶段肝细胞增生活跃,而虎眼万年青总皂苷干预此阶段的肝细胞增生。肝癌变过程中在肝癌发生阶段Flt-1表达较高,虎眼万年青总皂苷干预此阶段Flt-1表达。更多还原

【Abstract】 Objective:To study the anti-tumor effect of Ornithogalum Caudatum Ait total saponin(OCA-TS) on living diethylnitrosamine(DEN) induced hepatocarcinogenesis rat.Methods:the OCA-TS was administeredto interfere the hepatocarcinogenesis induced with DEN in rats,and the dynamic expression of pathological evolution,antioxidative activities of SOD,GSH-Px and the content of MDA were investigated by the colorimetric method in rat serum. Ki67 and Flt-1 were observed with immunohistochemical staining respectively.Results,As a result the procession of hepatocarcinogenesis in this model included three stages-hepatic toxic lesion、hepatic proliferation/ cirrhosis and hepatic carcinogenesis.Hyperplasia node,Basophilic cell lesions,lucencied and watering denaturalization appeared in the 12 weeks model group;a lot of foci-in-foci phenomenon appeared in the 14 weeks model group;carcinoma node appeared in the 18 weeks model group.Sprinkle basophilic cell lesions and fake flocculi,fibre alternation,local denaturalization appeared in the 12 weeks prevent group with OCA-TS;fake flocculi,abnormity fiber alternation,local watering and fatting denaturalization appeared in the 14 weeks prevent group with OCA-TS; segment hepatocirrhosis and gently denaturalization appeared in the 14 weeks treated group with OCA-TS.Hepatocirrhosis,hyperplasia node,local denaturalization and wrap around carcinoma with cyst appeared in the 18 weekstreated group with OCA-TS.Under the electronic microscope,part of vacuole,lipid droplet,cristae rupture,swelling,and metamorphose in the mitochondrion and endoplasmic reticulum expanded appeared in the 14 weeks model group.Some part of mitochondrion-endoplasmic reticulum complex appeared in the 14 weeks prevent group.Some part of vacuole,lipid droplet and cristae rupture appeared in the 14 weeks treated group,but there were no metamorphose in the mitochondrion and endoplasmic reticulum expand.Nuclear cataclasm,chromatin gather in the board,mitochondrion contract,endoplasmic reticulum swelling,and pull off granule appeared in the 18 weeks model group.Mitochondrion swelling,vacuole,lipid droplet,cristae rupture,endoplasmic reticulum expand,nuclear membrane metamorphose and chromatin distributing odds appeared in the chemotherapy group.Mitochondrion swelling and vacuole,endoplasmic reticulum expand with a lot of granule,cell plasma loosen appeared in the 18 weeks group treated with OCA-TS.The level of MDA increased remarkably in the 12 weeks、14 weeks and 18 weeks model group,and the content of MDA decreased by OCA-TS,there has significance between the groups administered with OCA-TS and the model groups respectively.In the 12 weeks model group the activities of SOD increased remarkably,but it tend to normal group in the 14 weeks,18 weeks model group.The expression of SOD in the 12 weeks prevent group administered with OCA-TS were lower than the 12weeks model group,maybe it dependent the level of MDA.The expression of GSH-Px activities decreased gradually in the 12 weeks、14 weeks and 18 weeks model group,and there have significance among of them.The expression of GSH-Px increased in the group administered with OCA-TS,and there have significance between the same periods model group respectively.We found the expression of Ki67 labeling index(LI) were significantly higher in the group of experiment than the normal group.The expression of Ki67 LI were highest in the 12 weeks model group,the OCA-TS interfered the expression of Ki67 LI in this periods significantly.And the Ki67 LI decreased gradually in the 14 weeks and 18 weeks model group,there have significant correlation between the 12 weeks model group respectively. And in these periods the OCA-TS interfered the expression of Ki67 LI not too much.We also found the expression of F1t-1 labeling index(LI) were significantly higher in the group of experiment than the normal group. The expression of Flt-1 LI were highest in the 14 weeks model group,the OCA-TS.interfered the expression of Flt-1 LI in this periods significantly. And the Flt-1 LI decreased in the 12 weeks and I8 weeks model group relatively,there have significant correlation between the 14 weeks model group respectively.And in these periods the OCA-TS interfered the expression of Flt-1 LI not too much.Conclusion:The procession of hepatocarcinogenesis interfered with OCA-TS appeared cyst wrap around carcinoma.OCA-TS interfered the hepatocarcinogenesis periods in the expression of the free radicals and antioxidants,and restrained the course of hepatocarcinogenesis.And the OCA-TS interfered the hepatocellular hyperplasia.The expression of Flt-1 were significantly high in the hepatocellular carcinoma occuring periods, and the OCA-TS interfered the expression of Flt-1 in this periods.更多还原