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目的:观察蝎毒多肽提取物(polypeptide extract from scorpion venom,PESV)对H22肝癌荷瘤小鼠化疗期间再增殖的抑制作用并探讨可能的机制。方法:96只Balb/c小鼠皮下接种小鼠肝癌H22细胞,随机分为模型组、PESV高、低剂量组和荷瘤对照组。以5-Fu对荷瘤小鼠进行化疗建立再增殖模型。按不同的用药方法对4组小鼠进行干预,每周测量肿瘤体积2次。各组每7 d处死6只小鼠,实验共进行35 d。以免疫组织化学方法检测各组肿瘤组织CD105,PCNA,VEGF,PDGF蛋白水平表达。以CD105标记微血管,计算微血管密度(MVD)。结果:荷瘤对照组肿瘤体积在13~24 d增加迅速,荷瘤对照组小鼠在第27天全部死亡,模型组肿瘤体积在17 d前增长较快,在第17~22天增加较慢,之后体积增加又较快,在第31天全部死亡。PESV高、低剂量组肿瘤体积全程缓慢增加,体积在17 d以后显著低于模型组,PESV高低剂量组之间仅在第17天存在差异(P<0.05)。免疫组织化学检测显示,模型组肿瘤组织第31天PCNA表达水平高于第21,28天(P<0.01),PESV高、低剂量组表达水平显著低于模型组(P<0.01),PESV高、低剂量组仅在第17天存在显著差异(P<0.05)。免疫组织化学检测显示,与PESV高、低剂量组相比,模型组CD105-MVD在第21,28天(P<0.05),第35天(P<0.01)存在显著差异,PESV高低剂量组之间无差别。模型组VEGF表达第35天高于第21,28天(P<0.05),PESV高、低剂量VEGF表达在第21,28,35天表达水平均显著低于模型组(P<0.01),PESV高低剂量组无差别。模型组肿瘤PDGF表达水平逐渐下降,PESV高,低组在第21天表达水平最低,之后逐渐增高,PESV高低剂量组之间在第35天存在显著差异(P<0.01)。相关性分析显示,肿瘤组织VEGF表达水平与肿瘤组织CD105-MVD呈正相关(r=0.669)。结论:PESV可抑制H22肿瘤在化疗期间再增殖作用,其机制可能是抑制血管生成和使肿瘤血管正常化。
Objective: To observe the inhibitory effect of polypeptide extract from scorpion venom (PESV) on the re-proliferation of H22 hepatoma tumor-bearing mice during chemotherapy and to explore the possible mechanism. Methods: Ninety-six Balb / c mice were inoculated subcutaneously with H22 cells and randomly divided into model group, PESV high and low dose groups and tumor-bearing control group. The 5-Fu tumor-bearing mice were treated with chemotherapy to establish a repopulation model. Four groups of mice were intervened by different medication methods, and the tumor volume was measured twice a week. Six mice were sacrificed every 7 days in each group for 35 days. The expression of CD105, PCNA, VEGF and PDGF in tumor tissue of each group was detected by immunohistochemical method. Microvessels were labeled with CD105 and microvessel density (MVD) was calculated. Results: The tumor volume of the tumor-bearing control group increased rapidly from 13 to 24 days. The tumor-bearing control mice all died on the 27th day. The tumor volume of the model group increased rapidly before 17 days and increased slowly from the 17th to the 22nd , Then increased in volume and faster, died on the 31st day. The volume of tumors in PESV high and low dose groups increased slowly throughout the whole process. The volume of PESV was significantly lower than that of model group after 17 days. There was difference between PESV high and low dose groups only on the 17th day (P <0.05). Immunohistochemistry showed that the expression level of PCNA on the 31st day in model group was higher than that on the 21st and 28th day (P <0.01), and the expression level of PESV in high and low dose group was significantly lower than that of model group (P <0.01) , There was significant difference only in the low dose group on the 17th day (P <0.05). Immunohistochemistry showed that CD105-MVD in model group was significantly higher than that in PESV high and low dose groups on day 21 and 28 (P <0.05) and 35 days (P <0.01) No difference between. The expression of VEGF in the model group was higher than that in the model group on the 35th day (P <0.05), and the expression of PESV in the high, low dose on the 21st, 28th and 35th day was significantly lower than that in the model group (P <0.01) No difference between high and low dose group. The expression of PDGF in the model group decreased gradually. The expression of PDGF in the high and low PESV groups was the lowest on the 21st day and then gradually increased. The difference between the PESV high and low dose groups on the 35th day was significant (P <0.01). Correlation analysis showed that the expression of VEGF in tumor tissue was positively correlated with CD105-MVD (r = 0.669). Conclusion: PESV can inhibit the proliferation of H22 tumor during chemotherapy. Its mechanism may be to inhibit angiogenesis and normalize the blood vessels of tumor.