Chan Shih-Feng, Chen Ya-Yin, Lin Jen-Jyh, Liao Ching-Lung, Ko Yang-Ching, Tang Nou-Ying, Kuo Chao-Lin, Liu Kuo-Ching, Chung Jing-Gung
Department of Biological Science and Technology, China Medical University, Taichung 404, Taiwan.
Department of Chinese-Western Medicine Integration, Chung Shan Medical University Hospital, Taichung 402, Taiwan.
Environ Toxicol. 2017 Feb;32(2):550-568. doi: 10.1002/tox.22259. Epub 2016 Mar 18.
Triptolide, a traditional Chinese medicine, obtained from Tripterygium wilfordii Hook F, has anti-inflammatory, antiproliferative, and proapoptotic properties. We investigated the potential efficacy of triptolide on murine leukemia by measuring the triptolide-induced cytotoxicity in murine leukemia WEHI-3 cells in vitro. Results indicated that triptolide induced cell morphological changes and induced cytotoxic effects through G0/G1 phase arrest, induction of apoptosis. Flow cytometric assays showed that triptolide increased the production of reactive oxygen species, Ca release and mitochondrial membrane potential (ΔΨ ), and activations of caspase-8, -9, and -3. Triptolide increased protein levels of Fas, Fas-L, Bax, cytochrome c, caspase-9, Endo G, Apaf-1, PARP, caspase-3 but reduced levels of AIF, ATF6α, ATF6β, and GRP78 in WEHI-3 cells. Triptolide stimulated autophagy based on an increase in acidic vacuoles, monodansylcadaverine staining for LC-3 expression and increased protein levels of ATG 5, ATG 7, and ATG 12. The in vitro data suggest that the cytotoxic effects of triptolide may involve cross-talk between cross-interaction of apoptosis and autophagy. Normal BALB/c mice were i.p. injected with WEHI-3 cells to generate leukemia and were oral treatment with triptolide at 0, 0.02, and 0.2 mg/kg for 3 weeks then animals were weighted and blood, liver, spleen samples were collected. Results indicated that triptolide did not significantly affect the weights of animal body, spleen and liver of leukemia mice, however, triptolide significant increased the cell populations of T cells (CD3), B cells (CD19), monocytes (CD11b), and macrophage (Mac-3). Furthermore, triptolide increased the phagocytosis of macrophage from peripheral blood mononuclear cells (PBMC) but not effects from peritoneum. Triptolide promoted T and B cell proliferation at 0.02 and 0.2 mg/kg treatment when cells were pretreated with Con A and LPS stimulation, respectively; however, triptolide did not significant affect NK cell activities in vivo. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 550-568, 2017.
雷公藤甲素是一种从雷公藤中提取的传统中药，具有抗炎、抗增殖和促凋亡特性。我们通过检测雷公藤甲素在体外对小鼠白血病WEHI-3细胞的细胞毒性，研究了其对小鼠白血病的潜在疗效。结果表明，雷公藤甲素诱导细胞形态变化，并通过G0/G1期阻滞诱导细胞毒性作用，引发细胞凋亡。流式细胞术检测显示，雷公藤甲素增加了活性氧的产生、钙离子释放和线粒体膜电位（ΔΨ），并激活了半胱天冬酶-8、-9和-3。雷公藤甲素增加了WEHI-3细胞中Fas、Fas-L、Bax、细胞色素c、半胱天冬酶-9、Endo G、Apaf-1、PARP、半胱天冬酶-3的蛋白水平，但降低了AIF、ATF6α、ATF6β和GRP78的水平。基于酸性空泡的增加、单丹磺酰尸胺对LC-3表达的染色以及ATG 5、ATG 7和ATG 12蛋白水平的增加，雷公藤甲素刺激了自噬。体外数据表明，雷公藤甲素的细胞毒性作用可能涉及凋亡和自噬相互作用之间的串扰。将正常BALB/c小鼠腹腔注射WEHI-3细胞以诱发白血病，然后分别以0、0.02和0.2 mg/kg的剂量口服雷公藤甲素进行治疗，持续3周，之后对动物进行称重，并采集血液、肝脏和脾脏样本。结果表明，雷公藤甲素对白血病小鼠的体重、脾脏和肝脏重量没有显著影响，然而，雷公藤甲素显著增加了T细胞（CD3）、B细胞（CD19）、单核细胞（CD11b）和巨噬细胞（Mac-3）的细胞数量。此外，雷公藤甲素增加了外周血单个核细胞（PBMC）中巨噬细胞的吞噬作用，但对腹膜巨噬细胞没有影响。当细胞分别用刀豆蛋白A和脂多糖刺激预处理时，雷公藤甲素在0.02和0.2 mg/kg剂量下促进了T细胞和B细胞的增殖；然而，雷公藤甲素在体内对NK细胞活性没有显著影响。© 2016威利期刊公司。《环境毒理学》32: 550 - 568, 2017。
