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目的:从绿茶中分离纯化阿拉伯半乳糖蛋白(AGPs),对其肝脏靶向性进行初步研究。方法:将脱色后的绿茶粉末在80?℃加水(1:20,w/v)浸提1.5h,获得的上清浓缩后用乙醇沉淀,收集重新溶解后在离子交换色谱Q Sepharose Fast Flow柱(20mm×60cm)和SephadexG100柱(16mm×60cm)纯化,收集主要含糖组分,冷冻干燥后获得绿茶AGPs,对其进行单糖组成、氨基酸组成、蛋白含量、糖醛酸含量分析。分别以200、400和800mg/kg.d3种剂量的绿茶AGPs灌胃昆明小鼠,2周后处死,取肝脏测定湿重和肝糖元含量。结果:分离纯化获得的绿茶AGPs分子量约为100kDa,是一种主要单糖组成为阿拉伯糖和半乳糖、含有一定量糖醛酸、蛋白含量低于10%的水溶性糖蛋白。初步研究发现绿茶AGPs与其它来源的AGPs一样具有肝脏靶向性。结论:绿茶AGPs的安全性和肝脏靶向性使其在药品输送上具有良好的应用前景。
OBJECTIVE: To isolate and purify arabinogalactan proteins (AGPs) from green tea and to study their liver targeting. Methods: The decolored green tea powder was extracted with water (1:20, w / v) at 80 ℃ for 1.5h. The obtained supernatant was concentrated, precipitated with ethanol, collected, reconstituted and purified by ion exchange chromatography on a Q Sepharose Fast Flow column (20mm × 60cm) and SephadexG 100 (16mm × 60cm). The main sugar components were collected and freeze-dried to obtain green tea AGPs. The monosaccharide composition, amino acid composition, protein content and uronic acid content were analyzed. Kunming mice were inoculated with green tea AGPs at doses of 200, 400 and 800 mg / kg.d respectively. Two weeks later, the mice were sacrificed and the wet weight and content of hepatic glycogen were measured. Results: The purified green tea AGPs obtained by isolation and purification had a molecular weight of about 100 kDa. It is a water-soluble glycoprotein whose major monosaccharide is composed of arabinose and galactose, contains a certain amount of uronic acid, and has a protein content of less than 10%. Preliminary studies have found that green tea AGPs, like other AGPs, have liver targeting. Conclusion: The safety of green tea AGPs and liver targeting make it have a good prospect of drug delivery.