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Abstract：
Cassia angustifolia Vahl. plant is used for many therapeutic purposes, for example, in people with constipation, skin diseases, including helminthic and parasitic infections. In our study, we demonstrated an amoebicidal activity of C. angustifolia extract against Acanthamoeba triangularis trophozoite at a micromolar level. Scanning electron microscopy (SEM) images displayed morphological changes in the Acanthamoeba trophozoite, which included the formation of pores in cell membrane and the membrane rupture. In addition to the amoebicidal activity, effects of the extract on surviving trophozoites were observed, which included cyst formation and vacuolization by a microscope and transcriptional expression of Acanthamoeba autophagy in response to the stress by quantitative polymerase chain reaction. Our data showed that the surviving trophozoites were not transformed into cysts and the trophozoite number with enlarged vacuole was not significantly different from that of untreated control. Molecular analysis data demonstrated that the mRNA expression of AcATG genes was slightly changed. Interestingly, AcATG16 decreased significantly at 12 h post treatment, which may indicate a transcriptional regulation by the extract or a balance of intracellular signalling pathways in response to the stress, whereas AcATG3 and AcATG8b remained unchanged. Altogether, these data reveal the anti-Acanthamoeba activity of C. angustifolia extract and the autophagic response in the surviving trophozoites under the plant extract pressure, along with data on the formation of cysts. These represent a promising plant for future drug development. However, further isolation and purification of an active compound and cytotoxicity against human cells are needed, including a study on the autophagic response at the protein level.
摘要：
决明子.植物用于许多治疗目的，例如,便秘的人，皮肤病,包括蠕虫和寄生虫感染。在我们的研究中,我们证明了在微摩尔水平上对棘阿米巴三角滋养体的杀阿米巴活性。扫描电子显微镜（SEM）图像显示了棘阿米巴滋养体的形态变化，其中包括细胞膜上孔隙的形成和膜破裂。除杀变形虫活性外,观察提取物对存活滋养体的影响，其中包括显微镜下的囊肿形成和空泡化，以及通过定量聚合酶链反应响应应激的棘阿米巴自噬的转录表达。我们的数据表明，存活的滋养体没有转化为囊肿，并且液泡扩大的滋养体数量与未处理的对照没有显着差异。分子分析数据表明AcATG基因的mRNA表达略有变化。有趣的是,AcATG16在治疗后12小时显着降低，这可能表明提取物的转录调节或细胞内信号通路的平衡，以响应压力，而AcATG3和AcATG8b保持不变。总之,这些数据揭示了沙棘提取物的抗棘阿米巴活性和植物提取物压力下存活的滋养体的自噬反应，以及囊肿形成的数据。这些代表了未来药物开发的有希望的植物。然而,需要进一步分离和纯化活性化合物以及对人细胞的细胞毒性，包括对蛋白质水平的自噬反应的研究。
