Studies on the mechanisms of the anti-inflammatory and anti-viral native substances

天然抗炎抗病毒物质的作用机制研究

基本信息

批准号：
08670532
负责人：
OHTSUKI Kenzo
金额：
$ 1.41万
依托单位：
Kitasato University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

The project study was done to investigate (i) purification of the specific binding proteins (gbPs) of anti-inflammatory egent, glycyrrhizin, from GL-sensitive mammalian cells, (ii) identification of gbPs by determination of their N-terminal partial amino acid sequences, and (iii) kinetics of the GL-induced selective inhibition of their physiological activities in vitro.To purify the gbPs from GL-sensitive mammalian cells or mouse tissues (liver and spleen) , a GL-affinity column was prepared. By using this GL-affinity column, several gbPs were purified to homogenous polypeptides after partial purification of the 1.5 M NaC1 clude extracts by combination with CM-cellulose column cromatography and gel filtration on a Superdex 200pg column (HPLC) . Determination of their N-termina partial amino acid sequences revealed that (i) arachidonate cascade enzymes, such as phospholipase A_2 (PLA_2) , 5'-1ipoxygenase (5'-LOX) and lipocortin I,are identified as gbPs ; (ii) serine protease (p32) , hyaluronidase (p55) comperiment C3 and immunoglobulin (Igg) are identified as gbPs in human serum ; and (iii) 14 kDa secetory PLA_2 (14 kDa sPLA_2 ) and serine protease are identified as gbPs in the synovial fluids of patients with rheumatonoid arthritis. Furthermore, kenitic studies revealed that GL,but not GA,at low levels (1-5muM) inhibits the-physiological activities of these gbPs and also selectively inhibits the CK-II-mediated activation of these gbPs in vitro.

进行项目研究是为了研究（i）（i）从GL对GL敏感的哺乳动物细胞中纯化抗炎eTIGT的特定结合蛋白（GBP），（II）通过确定其N-末端部分氨基酸序列和（III）KINETIFES造成的质量促进性的质量，以鉴定GBPS的质量促进性的质量促进性促进性的质量。从对GL敏感的哺乳动物细胞或小鼠组织（肝脏和脾脏）中，制备了GL-亲和力柱。通过使用此GL-亲和力柱，通过与CM-纤维素柱曲线曲线和SuperDex 200PG柱（HPLC）上的CM-纤维素柱曲线和凝胶过滤结合对1.5 M NaC1克拉德提取物进行部分纯化后，将几个GBP纯化为均匀的多肽。其N末端部分氨基酸序列的确定表明（i）蛛网膜级联酶，例如磷脂酶A_2（PLA_2），5'-1 ipoxygengenase（5'-lox）和脂皮质素I，被鉴定为GBPS；（ii）丝氨酸蛋白酶（p32），透明质酸酶（p55）成分C3和免疫球蛋白（IgG）在人血清中被鉴定为Gbps；（iii）14 kDa secetory PLA_2（14 kDa spla_2）和丝氨酸蛋白酶在类风湿关节炎患者的滑液中被鉴定为GBPS。此外，Kenitic的研究表明，在低水平（1-5MUM）下进行的GL（1-5MUM）抑制了这些GBP的生理活性，并且在体外也有选择地抑制CK-II介导的这些GBP的激活。