三氧化二砷对细粒棘球蚴原头节生长的影响及作用机制

发布时间：2018-06-16 18:48

本文选题：细粒棘球蚴 + 体外实验　；参考：《石河子大学》2015年硕士论文

【摘要】：目的:目前对包虫病的治疗,手术仍然是主要的手段,但术中易造成包虫囊肿囊液外溢,以及未杀死的原头节残留于手术创面或腹腔,这是导致手术后复发最主要的原因,因此术中、术后应用短时间内杀死头节又对周围正常组织损伤小的药物是阻止该病复发最常用的方法。本研究以体外培养的细粒棘球蚴原头节为研究对象,观察三氧化二砷对绵羊细粒棘球蚴原头节生长的影响,进一步评价三氧化二砷用于临床抗包虫治疗的可能性,为三氧化二砷在肝包虫病治疗的临床应用提供依据。方法:将新鲜的自然感染细粒棘球蚴的绵羊肝脏带回实验室,用无菌方式抽取囊内容物置培养瓶中,然后体外培养原头节3天,待适应体外环境后分别用含2μmol/L、4μmol/L、6μmol/L、8μmol/L浓度的三氧化二砷作用。用0.1%的伊红溶液染色,在光学显微镜下观察三氧化二砷不同时间、不同浓度对原头节的活力的影响(死的原头节会被染成红色),实验单独重复三次,根据数值绘制活力曲线。SEM观察原头节表面超微结构;TEM观察原头节内部超微结构的改变;用Caspase-3试剂盒检测原头节内Caspase-3酶活性;Western blot检测原头节p-Akt蛋白表达水平。结果:体外孵育14d后,空白1640对照组原头节的活力几乎没有改变。三氧化二砷各浓度组均对原头节有杀伤作用,8μmol/L三氧化二砷浓度组杀伤作用强于其他浓度组。表面超微结构出现凹陷,原头节呈外翻,顶突破坏,内部超微结构变化有合胞体带变薄、结构松散,出现少量脂滴,微绒毛减少。各药物作用组Caspase-3活性较正常对照组活性增加。Western blot结果显示P-Akt的表达量随药物浓度及时间的增加而呈明显下降趋势。结论:1.三氧化二砷对细粒棘球蚴原头节有杀伤作用。2.其作用机制可能是抑制了细粒棘球蚴原头节中p-Akt的表达,并可上调Caspase-3的表达而抑制生长、促进凋亡。
[Abstract]:Objective: at present, the treatment of hydatid disease is still the main method, but it is easy to cause hydatid cyst fluid spillover, and the non killed original joint is residual in the surgical wound or abdominal cavity, which is the main cause of recurrence after operation. Therefore, during the operation, the operation of the head joint in short time after operation is small in the surrounding normal tissue injury. The drug is the most commonly used method to prevent the recurrence of the disease. In this study, the effect of arsenic trioxide on the growth of echinococcosis in sheep was observed in vitro, and the possibility of arsenic trioxide used in the treatment of echinococcosis was further evaluated, and the treatment of arsenic trioxide in the liver hydatid disease was in the face of the treatment of liver hydatid disease. Methods: the sheep liver of the fresh and natural Echinococcus granulosus was brought back to the laboratory, and the capsule content was extracted with aseptic method, and then the original joint was cultured in vitro for 3 days. After the environment was adapted to the environment, the arsenic trioxide containing 2 mol/L, 4, mol/L, 6 u mol/L and 8 mu mol/L was used respectively. 0.1% Yi Hong was used. The effect of arsenic trioxide at different time and different concentration on the vitality of the original joint was observed under the optical microscope (the original joint was dyed red). The experiment was repeated three times alone, and the surface ultrastructure of the original joint was observed by the numerical activity curve.SEM. The changes of the inner ultrastructure of the original joint were observed by TEM; Caspase-3 The Caspase-3 enzyme activity in the original joint was detected by the kit and the expression level of the p-Akt protein was detected by Western blot. Results: after incubating for 14d in vitro, the activity of the original joint in the blank 1640 control groups was almost unchanged. The concentrations of arsenic trioxide all had a killing effect on the original joint, and the killing effect of the 8 micron arsenic trioxide concentration group was stronger than the other concentration. The surface ultrastructure appeared sunken, the original joint was ectropion, the apical process was destroyed, the inner ultrastructure changed with the syncytial body thinning, the structure loosened, a small amount of lipid droplets appeared, and the microvilli decreased. The activity of Caspase-3 in each drug group was increased by.Western blot results, which showed that the expression of P-Akt was increased with the increase of drug concentration and time. Conclusion: 1. arsenic trioxide has a killing effect on Echinococcus granulosus (Echinococcus granulosus).2., which may inhibit the expression of p-Akt in Echinococcus granulosus, and increase the expression of Caspase-3 to inhibit growth and promote apoptosis.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R657.3

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