人红细胞保护蛋白提纯及其对葡萄膜炎的作用  

Purification of protector protein from human red blood cells and its effects on uveitis

作　　者：金浩丽[1] 杨培增[1] 顾熊飞[2] 邢琳[1] 马翠萍[3] 周红颜[1] 黄祥坤[1] 谢楚芳[1] 

机构地区：[1]中山大学中山眼科中心,广州510060 [2]中山大学 [3]中山大学第一附属医院

出　　处：《眼科研究》2003年第5期497-500,共4页Chinese Ophthalmic Research

基　　金：国家自然科学基金(30171002)

摘　　要：目的 探讨人红细胞保护蛋白(PRP)对内毒素诱导葡萄膜炎(EIU)的影响。方法 用等量饱和硫酸铵溶液从人新鲜红细胞中盐析出基质可溶性蛋白,进行CM-Sephadex C50阳离子交换和DEAE Sephacel阴离子交换层析,以提纯PRP。6只PRP处理SD大鼠和4只未处理SD大鼠均于足底部注射200μg鼠伤寒杆菌内毒素(LPS),以诱导EIU,每只PRP处理鼠在LPS注射同时及注射前2h腹腔注射0.25mg还原型PRP,于LPS注射后24h评价两组大鼠临床表现、房水蛋白浓度、房水细胞数及虹膜睫状体平片上ED1^+细胞数等。结果 从30ml人新鲜红细胞中可提纯15.3mg的PRP,其纯度为99％;PRP处理组大鼠在临床表现积分、房水蛋白质量浓度、房水细胞数和虹膜睫状体平片上ED1^+细胞数等方面都显著低于未处理组(P<0.05)。结论 用CM-Sephadex C50阳离子交换和DEAE Sephacel阴离子交换层析的方法能从红细胞中提取高产量、高纯度的PRP;PRP对EIU有预防和治疗作用。Objective To purify protector protein (PRP) from human red blood cells ( HRBC) and to study its effects on endotoxin-induced uveitis (EIU). Methods PRP was purified by subjecting the soluble protein from HRBC to CM-Sephadex C50 column and DEAE-Sephacel column. Salmonella Typhimurium Lipoplysaccharide (LPS) of 100ug was injected into each hind footpad of 6 PRP-treated Spar-Dawley (SD) rats and 4 untreated SD rats with a total dose of 200 ug LPS for each animal. Reduced PRP was injected at a dosage of 0. 25 mg intraperitoneally into each PRP-treated rat at 2 h before injection and at the same time of LPS injection. Parameters of inflammatory scales, protein concentration and inflammatory cells in the aqueous humor and ED1+ cells in the iris-ciliary body were evaluated at 24 h after LPS injection. Results A total amount of 15. 3 mg purified PRP was obtained from 30 ml HRBC, and its purity was over 99%. Decreased inflammation, lower protein concentration and fewer inflammatory cells in the aqueous humor and ED1+ cells in the iris-ciliary body were observed in the PRP-treated rats as compared with the untreated ones. Conclusions High production and high purity PRP can be obtained from HRBC with the method of two-step chromatography. The study reveals that PRP is effective in reducing the intraocular inflammation induced by endotoxin.

关 键 词：人红细胞保护蛋白 葡萄膜炎 提取纯化 内毒素 大鼠 

分 类 号：R773.9[医药卫生—眼科]