日本血吸虫体表特异性结合分子MppZL4抗虫性的研究
etection to the Killing-worm Function of Schitosoma Japonicum-binding MppZL4
目的:通过 MppZL4在体内外对S.j细胞及虫体的杀伤实验,检测MppZL4的抗虫性功能。方法:小鼠感染 S.j 24d 后,尾静脉注射MppZL4 1012pfu,15 min 后剖杀小鼠取出虫体制备 S.j 细胞,免疫细胞化学检测MppZL4在细胞上的定位;体外培养MppZL4处理的S.j细胞1d后,MTT试剂盒检测其抑制率,计算 IC50。MppZL4处理S.j脱尾童虫,每24h美蓝染色观察一次,连续观察72 h,统计虫体死亡率。小鼠感染S.j 42d后经尾静脉注射MppZL4 1012pfu 连续7天,取小鼠肝脏,免疫组织化学检测MppZL4对S.j虫卵的作用。小鼠感染血吸虫静脉注射MppZL4 1011pfu,连续注射7d,50d后常规灌注冲虫、观察鼠肝并作虫卵计数。以上实验,均设置阳性对照、阴性对照及空白对照组。结果:免疫细胞化学证实,MppZL4主要沉积于 S.j大圆细胞、三角形细胞及不规则细胞的细胞膜上。MTT检测显示,MppZL4浓度为 1010pfu时,具有明显的细胞毒性(P=0.000),且随着MppZL4浓度的增高,毒性效应增强;IC50≈1010。体外杀伤实验观察到MppZL4致死率高于东方田鼠血清阳性对照组(P=0.000);48 h后,其死亡率已达到100%。免疫组织化学检测MppZL4作用感染小鼠后,肝脏内虫卵发育不全,毛蚴结构模糊。动物实验显示MppZL4治疗组肝脏较为柔软,红润,虫卵结节较少,未见明显结缔组织形成;减虫率为25.80%(P=0.010),减卵率为59.24%(P=0.000),虫体重量降低率为34.12%(P=0.000)。结论:MppZL4具有一定的抗血吸虫性。
Objective: The killing-worms functions of MppZL4were detected by the experiments in vivo and vitro. Method: The mice, infected with S.j 24th day, were injected with MppZL4 1012 pfu through tail vein and sacrificed after 15 min. The S.j cells were manufactured used the worms from the mice in sterile condition and detected MppZL4 by immunocytochemistry. The S.j cells were treated by MppZL4 for one day whose inhibition rate was studied by MTT kit through IC50. S.j schistosomula were treated with MppZL4 in axenic condition and observated every 24h for 72h through methylene blue stain, while the worms' mortality was counted. The mice, infected with S.j 42nd day, were injected with MppZL4 1012 pfu through tail vein for 7 days and sacrificed; The livers were removed and detected effect of MppZL4 on wroms by immunohistochemical test. The mice, infected with S.j , were injected with MppZL4 1011 pfu through tail vein for 7 days and sacrificed 50th day. The lives were removed and observed while the worms were collected and counted. The positive control, negative control and blank control group were established in all the tests. Result: Immunocytochemical stain displaied that MppZL4 phage mainly deposited the cell envelope of big round cells, triquetrous cells and irregular cells of Sj. MTT test illustrateed that MppZL4 1010 pfu possessed significant cytotoxicity (P=0.000), and with the higher concentration of MppZL4, the cytotoxicity enhanced; IC50≈1010. The death rate of MppZL4 lethal effect in vitro is higher than fortis serum positive control group (P=0.000) and its mortality rate had reached 100% after 48h. The results of immunohistochemical test show that the eggs in the lives of the infected mice were hypoplastic and fuzzy.Animal experiments had shown, that the liver of MppZL4 treatment group soft, rosy, fewer eggs nodules, no significant formation of connective tiss, relatively. The rate of worm burden reduction was 25.80% (P=0.010), LEPG reduction was 59.24%(P=0.000), and worm weight reduction was 34.12% (P=0.000). Conclusion: The specific MppZL4 possess killing-worms function to S.j.
刘彦、曾庆仁、杨胜辉、邹菊、梁瑜、陈莉
基础医学微生物学分子生物学
日本血吸虫MppZL4抗虫性
Schistosoma japonicumMppZL4killing-worms function
刘彦,曾庆仁,杨胜辉,邹菊,梁瑜,陈莉.日本血吸虫体表特异性结合分子MppZL4抗虫性的研究[EB/OL].(2015-06-09)[2025-08-18].http://www.paper.edu.cn/releasepaper/content/201506-123.点此复制
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