本文選題：細粒棘球蚴 + 體外實驗��； 參考：《石河子大學》2015年碩士論文

【摘要】：目的:目前對包蟲病的治療,手術(shù)仍然是主要的手段,但術(shù)中易造成包蟲囊腫囊液外溢,以及未殺死的原頭節(jié)殘留于手術(shù)創(chuàng)面或腹腔,這是導致手術(shù)后復發(fā)最主要的原因,因此術(shù)中、術(shù)后應用短時間內(nèi)殺死頭節(jié)又對周圍正常組織損傷小的藥物是阻止該病復發(fā)最常用的方法。本研究以體外培養(yǎng)的細粒棘球蚴原頭節(jié)為研究對象,觀察三氧化二砷對綿羊細粒棘球蚴原頭節(jié)生長的影響,進一步評價三氧化二砷用于臨床抗包蟲治療的可能性,為三氧化二砷在肝包蟲病治療的臨床應用提供依據(jù)。方法:將新鮮的自然感染細粒棘球蚴的綿羊肝臟帶回實驗室,用無菌方式抽取囊內(nèi)容物置培養(yǎng)瓶中,然后體外培養(yǎng)原頭節(jié)3天,待適應體外環(huán)境后分別用含2μmol/L、4μmol/L、6μmol/L、8μmol/L濃度的三氧化二砷作用。用0.1%的伊紅溶液染色,在光學顯微鏡下觀察三氧化二砷不同時間、不同濃度對原頭節(jié)的活力的影響(死的原頭節(jié)會被染成紅色),實驗單獨重復三次,根據(jù)數(shù)值繪制活力曲線。SEM觀察原頭節(jié)表面超微結(jié)構(gòu);TEM觀察原頭節(jié)內(nèi)部超微結(jié)構(gòu)的改變;用Caspase-3試劑盒檢測原頭節(jié)內(nèi)Caspase-3酶活性;Western blot檢測原頭節(jié)p-Akt蛋白表達水平。結(jié)果:體外孵育14d后,空白1640對照組原頭節(jié)的活力幾乎沒有改變。三氧化二砷各濃度組均對原頭節(jié)有殺傷作用,8μmol/L三氧化二砷濃度組殺傷作用強于其他濃度組。表面超微結(jié)構(gòu)出現(xiàn)凹陷,原頭節(jié)呈外翻,頂突破壞,內(nèi)部超微結(jié)構(gòu)變化有合胞體帶變薄、結(jié)構(gòu)松散,出現(xiàn)少量脂滴,微絨毛減少。各藥物作用組Caspase-3活性較正常對照組活性增加。Western blot結(jié)果顯示P-Akt的表達量隨藥物濃度及時間的增加而呈明顯下降趨勢。結(jié)論:1.三氧化二砷對細粒棘球蚴原頭節(jié)有殺傷作用。2.其作用機制可能是抑制了細粒棘球蚴原頭節(jié)中p-Akt的表達,并可上調(diào)Caspase-3的表達而抑制生長、促進凋亡。
[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.
【學位授予單位】：石河子大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R657.3

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