Sulfate conjugation of daphnetin by the human cytosolic sulfotransferases.
Sulfate conjugation of daphnetin by the human cytosolic sulfotransferases.
复制标题
DOI：
10.1016/j.jep.2016.05.041
复制
发表时间：
2016-08-02
影响因子：
5.4
通讯作者：
Liu MC
中科院分区：
文献类型：
--
作者：
Han Z;Xi Y;Luo L;Zhou C;Kurogi K;Sakakibara Y;Suiko M;Liu MC
在土耳其，含有达芙妮素的达芙妮油被用作治疗风湿性疼痛和腰痛的民间药物。中国有一种名为“足马片”的中药片剂，用于治疗类风湿关节炎。本研究旨在探讨在体外培养的人细胞中通过硫酸化对瑞香素的代谢，并鉴定能够介导瑞香素硫酸化的人细胞质硫转移酶（SULT）。培养的HepG2人肝癌细胞和Caco-2人结肠癌细胞在不同浓度的瑞香素存在下用[35S]硫酸盐标记。13种已知的人SULTS，先前表达和纯化，以及人肾、肝、肺和小肠的细胞质，使用已建立的硫转移酶测定法检测了石仙花硫酸化活性。[35S]硫酸盐标记的HepG2细胞和Caco-2细胞在马瑞素存在的情况下产生并释放[35S]硫酸马瑞素。在已知的13种人类SULTs中，SULT1A1、SULT1A2、SULT1A3、SULT1B1和SULT1C4对瑞香素表现出显著的硫酸酸化活性。在随后测试的四种人体器官样本中，小肠和肝脏细胞质显示出比肺和肾更高的硫酸化活性。本研究的结果明确表明，在培养的人类细胞中，纯化的人类SULT酶以及人类器官细胞质中，瑞香素可以被硫酸酸化。所得信息为进一步研究瑞香素在体内的硫酸化代谢提供了基础。
In Turkey, daphnetin-containing Daphne oleoides is used as a folk medicine for treating rheumatic pain and lumbago. A daphnetin-containing traditional Chinese medicine tablet, named Zushima-Pian, is available in China for treating rheumatoid arthritis. The present study aimed to investigate the metabolism of daphnetin through sulfation in cultured human cells and to identify the human cytosolic sulfotransferase(s) (SULT(s)) that is(are) capable of mediating the sulfation of daphnetin. Cultured HepG2 human hepatoma cells and Caco-2 human colon carcinoma cells were labeled with [35S]sulfate in the presence of different concentrations of daphnetin. Thirteen known human SULTS, previously expressed and purified, as well as cytosols of human kidney, liver, lung, and small intestine, were examined for daphnetin-sulfating activity using an established sulfotransferase assay. [35S]sulfated daphnetin was found to be generated and released by HepG2 cells and Caco-2 cells labeled with [35S]sulfate in the presence of daphnetin. Among the 13 known human SULTs, SULT1A1, SULT1A2, SULT1A3, SULT1B1, and SULT1C4 displayed significant sulfating activity toward daphnetin. Of the four human organ samples later tested, small intestine and liver cytosols displayed considerably higher daphnetin-sulfating activity than those of lung and kidney. The results derived from the present study showed unequivocally that daphnetin could be sulfated in cultured human cells and by purified human SULT enzymes as well as human organ cytosols. The information obtained provided a basis for further studies on the metabolism of daphnetin through sulfation in vivo.