葛根素注射液对家兔不良反应的实验研究

Puerarin Injection Induced Adverse Reaction in Rabbits

目的研究葛根素注射液的不良反应情况及作用机制。方法参考反映Ⅱ型变态反应的免疫毒理学研究模型,以家兔耳缘静脉注射绵羊红细胞为阳性对照组,连续注射葛根素注射液(28.6 mg.kg-1)6 d,停药1 d,持续6周,停药当天观察家兔免疫系统大体症状如呼吸困难、喷嚏等,并监测体温;检测血液、尿液中溶血相关指标和肝肾功能指标;用Coombs实验法检测家兔血清体外使红细胞溶血情况;给药结束后处死家兔测定脏器指数,并进行组织学观察。结果家兔注射葛根素注射液3周开始出现尿潜血,第5、6周血浆游离血红蛋白显著增加,提示家兔可能出现血管内溶血。中性粒细胞数目从1周开始减少,其中第4周与对照组相比有显著性差异(P<0.05),提示对粒细胞有损伤;淋巴细胞数目在第2周显著增加(P<0.05);血小板数目于第6周显著增加;脏器指数显示胸腺增生,提示免疫系统参与此过程。血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)及肝脏病理学结果提示肝脏损伤。而免疫系统大体反应均未见显著变化。Coombs试验结果表明,3周后有67%的家兔血清在体外使与葛根素抗原共孵育的红细胞溶血,提示其含有葛根素抗体。结论静脉注射葛根素注射液会引起家兔溶血、粒细胞减少、淋巴细胞增加及肝损伤等不良反应,其机制可能与Ⅱ型变态反应相关。

OBJECTIVE To explore the adverse reaction of puerarin injection and its mechanism in animal model.METHODS Rabbits were intravenously administrated with puerarin injection through the edge of ear for 6 weeks.Rabbits were injected with sheep erythrocytes as positive control.The reactions of immune system were observed,and the blood and urine were obtained every week.Blood was used to determine free hemoglobin,unconjugated bilirubin,alanine aminotransferase,urea nitrogen,granulofilocyte,neutrophil and platelet.Urine was used to analyze occult blood,urobilinogen and urine color.After 6 weeks,the rabbits were sacrificed to gain spleen,thymus gland,liver and kidney.Organ weights were measured and histopathologic examination was performed.RESULTS Occult blood in urine was obtained after the treatment of 3 weeks of sheep erythrocytes.After 5 weeks of puerarin treatment,free hemoglobin was obviously increased,suggesting intravascular hemolysis.Neutrophil was decreased significantly compared with control（P0.05）,while achroacyte was increased significantly（P0.05）,which indicated that immune system was involved.Alanine aminotransferase and histopathologic examination showed that the liver was injured.Coombs test showed that hemolysis were occurred with 67% of rabbit serum in sheep erythrocytes incubated with puerarin,which suggested that rabbit serum possessed the antibody of puerarin.CONCLUSION Puerarin injection would cause adverse reaction including intravascular hemolysis,granulocytopenia,achroacyte increasing and hepatic injury.The mechanism is involved in type Ⅱ allergy.