斯氏艾美球虫可溶性蛋白对小鼠结肠癌皮下肿瘤模型的影响

发布时间：2018-06-17 04:27

  本文选题：斯氏艾美球虫 + 结肠癌　； 参考：《中国寄生虫学与寄生虫病杂志》2017年04期

【摘要】：目的探讨斯氏艾美球虫(Eimeria stiedai)可溶性蛋白(EsSP)对荷瘤小鼠肿瘤生长、生存率和免疫状态的影响。方法建立小鼠皮下结肠癌(CT26)肿瘤模型,确定最小100%致瘤量肿瘤细胞数。取10~8个斯氏艾美球虫孢子化卵囊,采用超声波间断乳化制备EsSP。105只雄性BALB/c小鼠按随机数表法均分为7组(15只/组),每组小鼠于右侧腋窝皮下接种CT26细胞5×10~5个,其中6个实验组(A~F组)小鼠分别腹腔注射100.00、50.00、10.00、1.00、0.10、0.01μg/d EsSP,1次/d×5 d,对照组腹腔注射等量PBS。于接种后第7、11、13、15、17、19、21、23天测量肿瘤直径,计算相对肿瘤体积和相对肿瘤增殖率(T/C)。接种后第25天每组各处死5只小鼠,称量瘤重,计算抑瘤率。采眼球血,分离小鼠外周血淋巴细胞,MTS比色法检测EsSP对荷瘤小鼠淋巴细胞增殖能力的影响,计算刺激指数(SI);流式细胞术检测外周血CD4~+/CD8~+T淋巴细胞比值变化。记录各组荷瘤小鼠死亡时间和死亡数量,共观察80 d。各组间差异性比较采用单因素方差分析。结果最小100%致瘤量肿瘤细胞数为5×10~5个。接种结肠癌细胞后第23天,A、B、C组的肿瘤体积为(435.2±41.1)、(366.3±29.2)、(460.2±28.5)mm~3,较E、F和对照组的(761.2±33.2)、(810.4±38.4)、(865.2±35.3)mm~3生长缓慢(P0.05);A、B、C组的T/C分别为(39.0±6.7)%、(33.3±8.9)%、(35.0±8.1)%,均40%。B组小鼠瘤重为(1.109±0.432)g,低于对照组的(1.946±0.289)g(P0.05),抑瘤率最高,为(43.0±14.6)%。MTS比色法检测结果显示,B、C组小鼠SI分别为1.75±0.15、1.70±0.32,高于对照组的1.38±0.18(P0.05)。流式细胞术检测结果显示,A、B、C组小鼠外周血中CD4~+/CD8~+T淋巴细胞亚群的比值分别为1.58±0.24、1.74±0.22、1.61±0.16,与对照组的1.34±0.15比较,差异均有统计学意义(P0.01或P0.05)。荷瘤小鼠生存率结果显示,至第80天,B、C组各存活5只小鼠,高于对照组的2只(P0.05)。结论 EsSP能够抑制结肠癌皮下肿瘤的生长,提高荷瘤小鼠生存率,改变肿瘤诱导的免疫抑制状态,增强小鼠抗肿瘤免疫反应。
[Abstract]:Objective to investigate the effects of Eimeria stiedaii (Eimeria stiedaii) soluble protein EsSPs on tumor growth, survival rate and immune status in tumor-bearing mice. Methods the tumor model of murine subcutaneous colon cancer (CT26) was established, and the tumor cell number of the tumor was determined at least 100%. The spore oocysts of Eimeria skrjabini were collected and 105 male BALB / c mice were prepared by ultrasonic discontinuous emulsification. According to the random number table, 105 male BALB / c mice were divided into 7 groups with 15 oocysts in each group. CT26 cells were subcutaneously inoculated in the right armpit of each group with 5 脳 105 CT26 cells. Six experimental groups (group A) were intraperitoneally injected with 100.00m 50.0010. 00 (1.00) and 0.01 渭 g / d EsSPN once a day 脳 5 days, and the control group were injected with the same amount of PBSs intraperitoneally. The mice in the control group were intraperitoneally injected with the same amount of PBSs. The tumor diameter was measured on the 7th day after inoculation. The relative tumor volume and the relative tumor proliferation rate were calculated. On the 25th day after inoculation, 5 mice were killed in each group, the tumor weight was weighed and the tumor inhibition rate was calculated. The effect of EsSP on lymphocyte proliferation in tumor-bearing mice was determined by MTS colorimetric assay and flow cytometry was used to detect the ratio of CD _ 4 ~ / CD _ 8 ~ T lymphocytes in peripheral blood. The time and number of death of tumor-bearing mice were recorded and observed for 80 days. Single factor analysis of variance (ANOVA) was used to compare the differences among groups. Results the number of tumor cells was 5 脳 10 ~ 5. 鎺ョ�嶇粨鑲犵檶缁嗚優鍚庣�，

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