摘要
健康猪胎盘经酶解,提取,氯仿-正丁醇除杂蛋白与5'-磷酸二酯酶除核酸,再经透析,乙醇沉淀,DEAE-Dextran-Gel A-25层析制得纯品猪胎盘脂多糖(简称P-脂多糖)。分析结果表明,P-脂多糖是一种类酸性粘多糖与脂质的共价结合物,分子量约为75kD,分子中含11种单糖组成,其中半乳糖含量最高,其次为岩藻糖,氨基已糖和已糖醛酸。体外实验发现P-脂多糖能明显诱导小鼠淋巴细胞的增殖,强烈地促进淋巴细胞的DNA合成,刺激指数约为9.3,最适刺激浓度50～60μg/mL,最适刺激时间48小时,同时能明显促进淋巴细胞的蛋白质合成。实验还证实,P-脂多糖对脾淋巴细胞的增殖诱导作用并不是通过依赖于白间素-2(IL-2)途径,而是通过增强淋巴细胞IgM的表达,表明P-脂多糖是一种促B-淋巴细胞分裂剂。
Placenta powder from health pig placenta was suspended in Water and treated with enzyme. Protein in extract was removed by chloroform-n-butanol partition. The supernatant was treated with phosphodiesterase, then dialysised and precipitated by alcohol.The crude lipopolysaccharide was further purified by DEAE-dextran-gel A-25. The pure lipopolysaccharide obtained was identified. The results showed that it is composed of acid mucoitin and lipid and contains 11 kinds of sugars, mainly gal-actose, focuse, hexosamine and glucuronic acid. The molecular weight was 75000 by chromatographyThe lipopolysaccharide dramatically stimulated proliferation, DNA and protein synthesis of lymphocytes. The stimulatory index of DNA. synthesis was about 9.3. A maxium proliferation was induced by treatment of 50-60μg/mL of lipopolysaccharide for 48 hours. It was also demostrated that the lipopolysaccharide induced lymphocytes proliferation by an interleukin-2-independent pathway and by increasing expression of IgM in lymphocytes. Therefore the lipopolysaccharide from pig placenta is a poly clonal B-cell mitogen.
基金
国家自然科学基金3860685资助
关键词
猪胎盘脂多糖
淋巴细胞
生物制品
制备
药理
Lipopolysaccharide from pig placenta, Lymphocytes proliferation,Mitogen
