A Study on the Apoptosis Induction and Mechanisms in Human Leukemia Cells Induced by Atractylenolide I

• Main Author: 林子文
• Other Authors: Huang, R.L.
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/14833058202775563474

碩士 === 美和科技大學 === 健康與生技產業研究所 === 100 === Baizhu (Atractylodes macrocephala), which belongs to Asteraceae, is a perennial herb. Its roots have long been applied in traditional Chinese medicine. Recently, Baizhu has been used in treating chronic diseases and post-surgery symptoms, such as tiredness and vomit. Besides, Baizhu is believed to be effective in healing human gastrointestinal disorders. The related literatures also showed that Baizhu depicts several functions, such as anti-cancer, anti-oxidation, and inhibiting angiogenesis. In this study, we first isolate and extract Atractylenolide I, the most active component in Baizhu. Next, we explore the effectiveness of Atractylenolide I in suppressing the proliferation of leukemia and inducing its apoptosis. In our experiments, MTT Assay, microscopy observation, DNA gel electrophoresis, ELISA kit, and flow cytometry were used to observe apoptosis. In addition, Western Blot was used to detect the pathway of caspase-9, caspase-3, and the cutting of the role of DNA repair enzyme PARP protein, to study the intrinsic apoptosis condition of leukemia cells. Our experimental results show that for the Atractylenolide I treated human T cell leukemia cell line Jurkat and human myeloid leukemia cell line U937 after 12, 24 and 48 hours via MTT assay, the IC50 values are 44.67, 37.63, 24.44 g/mL, and 24.11, 14.91, 4.24 g/mL, respectively, indicating that Atractylenolide I does inhibit these two human leukemia cell growth with time and concentration dependency. In addition, microscopy observation also confirmed corresponding cell appearance changes, while cells fragments were produced via DNA gel electrophoresis, and the apoptosis rate increase was observed by ELISA kit, and the sub-G1 apoptosis peak elevation was confirmed by flow cytometry. Finally, the Western Blot analysis also confirmed the activation of promoting the upstream proteins caspase-9, downstream executor caspase-3 protein, as well as cutting the role of DNA repair enzyme PARP. Based on the above findings, Atractylenolide I appear to effective in inducing Jurkat and U937 of leukemia cells to apoptosis. Our preliminary results seem encouraging and may be applicable to future medicinal development for leukemia treatment.