Kok L D, Wong C K, Leung K N, Tsang S F, Fung K P, Choy Y M
Department of Biochemistry, Chinese University of Hong Kong, Shatin, New Territories.
Immunopharmacology. 1995 Jun;30(1):79-87. doi: 10.1016/0162-3109(95)00010-q.
Radix bupleuri, the root of Bupleuri spp., Chinese medicinal herbs used for the treatment of influenza, malaria and menstrual disorders, were extracted with hot water and separated into five different fractions (RB, RBI, RBII, RBIII and RBIV) by stepwise alcohol precipitation. One of these fractions, RBI, was then fractionated into RBIa and RBIb by gel filtration using G-100 Sephadex. These two fractions were further purified into RBIai, RBIaii and RBIbi, RBIbii fractions respectively by ion-exchange chromatography using DEAE-Sephadex. Each of these fractions is a heteropolymer consisting mainly of carbohydrate and varying proportions of protein and uronic acid. RBIaii was found to show strong anti-tumor activities in sarcoma-bearing mice. Mechanistic studies showed that RBIaii exhibited a potent activating effect on the cytotoxic activity of macrophages, NK and LAK cells against tumor cells. In addition, RBIaii could increase the number of tumor infiltrating lymphocytes (TILs) in the tumor site of WEHI-164-bearing mice. Furthermore, RBIaii could induce the release of interferon-gamma by lymphocytes in vitro.
柴胡，即柴胡属植物的根，是用于治疗流感、疟疾和月经失调的中药材，用热水提取后通过逐步乙醇沉淀法分离成五个不同的组分（RB、RBI、RBII、RBIII和RBIV）。然后，其中一个组分RBI通过使用G-100葡聚糖凝胶的凝胶过滤法进一步分离成RBIa和RBIb。这两个组分分别通过使用DEAE-葡聚糖凝胶的离子交换色谱法进一步纯化得到RBIai、RBIaii和RBIbi、RBIbii组分。这些组分中的每一个都是主要由碳水化合物以及不同比例的蛋白质和糖醛酸组成的杂聚物。发现RBIaii在荷肉瘤小鼠中显示出强大的抗肿瘤活性。机制研究表明，RBIaii对巨噬细胞、NK和LAK细胞对肿瘤细胞的细胞毒活性具有强大的激活作用。此外，RBIaii可以增加荷WEHI-164小鼠肿瘤部位的肿瘤浸润淋巴细胞（TIL）数量。此外，RBIaii可以在体外诱导淋巴细胞释放γ-干扰素。
