双黄连注射剂中黄芩苷致敏家兔后血清中特异性抗体的测定

Determination of specific antibodies in serum of rabbits sensitized by baicalin in shuanghuanglian injection

目的酶联免疫吸附法(ELISA)检测双黄连注射剂中小分子半抗原物质黄芩苷致敏家兔后血清中特异性抗体效价。方法取新西兰种家兔10只,分为空白组、黄芩苷组,每组各5只。自家兔耳缘动脉采血10 m L,制备致敏前血清。将被液相色谱-质谱-血清白蛋白结合系统检测的黄芩苷与家兔血清体外孵育,制成黄芩苷-血浆蛋白完全抗原。将此完全抗原以皮下注射的方式对家兔进行免疫,免疫次数共5次。最后一次免疫结束后,间隔1周时间,将其麻醉,用采血针自家兔腹主动脉采血制备黄芩苷-血浆蛋白全抗原的抗血清。用ELISA法检测抗血清的抗体效价(OD值);间接竞争ELISA法测定家兔抗血清的特异性抗体;采用兔免疫球蛋白E(Ig E)ELISA试剂盒检测各组家兔抗血清中Ig E抗体的含量。结果黄芩苷的抗体效价(OD)与阴性对照组的(OD)比值>2,因此说明黄芩苷具有致敏性。家兔体内产生的抗血清中的抗体具有一定的特异性。家兔体内明显产生了外源性Ig E型抗体。结论黄芩苷对家兔具有致敏性。

Objective To detect the specific antibody titers by ELISA in the small molecular semi-antigenic baicalin components of shuanghuanglian injection in the rabbits sensitized by baicalin. Methods Total 10 New Zealand rabbits were randomized into the the blank group and baicalin group,5 rabbits in each group. Blood（ 10 m L） was collected from auricular artery for serum preparation befroe sensitization. The serum was incubated in vitro with baicalin which detected by lc-ms-serume albumin binding system,preparing baicalin-serum protein complete antigen. Rabbits were immunized by subcutaneous injuection of complete antigen 5 times. The antiserum of baicalin-serum protein complete antigen was collected from abdominal aorta at a one week interval after the last immunizations. ELISA was used to determine the specific antibody titers（ OD value） of antiserum. The indirect competitive ELISA was used to determine the antiserum specificity. ELISA kit was used to detect the level of rabbit immunoglobulin（ Ig E） antibody in serum. Results The antibody titers ratio of baicalin and nagative control group was more than 2,so that the baicalin had sensitization. The antibody from antiserum in rabbits had somewhat specificity.The exogenous Ig E antibody was produced in rabbits clearly. Conculsion Baicalin has sensitization to rabbits.