CAJ | 학술논문

目的：用RBL-2H3细胞初步评价清开灵、血塞通注射液类过敏反应,为完善中药注射剂类过敏反应的筛选提供实验依据。方法 MTT法测定2种药物对细胞生长的IC50；采用不同浓度药物、C48/80或培养液分别刺激细胞30 min,检测上清液中组胺、β-氨基己糖苷酶释放率；另取刺激后的细胞,观察其脱颗粒率及超微结构变化。取ICR小鼠进行类过敏试验,以含伊文思蓝的药物或生理盐水单次尾静脉注射30 min,记录每组耳廓蓝染的动物数、蓝染总耳数及耳廓伊文思蓝渗出量。结果离体实验显示,清开灵、血塞通注射液的 IC50均为12.5μL/mL；2种药物在较高浓度均能促进细胞释放组胺和β-氨基己糖苷酶(P＜0.05,P＜0.01),呈一定剂量依赖性；形态上可见细胞表面绒毛减少,内部空泡状结构增加。小鼠类过敏试验显示,2种药物对动物耳蓝染率均为100%、伊文思蓝渗出量明显增多(P＜0.01)。离体与整体实验结果一致。结论清开灵、血塞通注射液具有潜在的致类过敏反应性,RBL-2H3细胞模型用于中药注射剂类过敏性评价具有一定的参考应用价值。
목적：용RBL-2H3세포초보평개청개령、혈새통주사액류과민반응,위완선중약주사제류과민반응적사선제공실험의거。방법 MTT법측정2충약물대세포생장적IC50；채용불동농도약물、C48/80혹배양액분별자격세포30 min,검측상청액중조알、β-안기기당감매석방솔；령취자격후적세포,관찰기탈과립솔급초미결구변화。취ICR소서진행류과민시험,이함이문사람적약물혹생리염수단차미정맥주사30 min,기록매조이곽람염적동물수、람염총이수급이곽이문사람삼출량。결과리체실험현시,청개령、혈새통주사액적 IC50균위12.5μL/mL；2충약물재교고농도균능촉진세포석방조알화β-안기기당감매(P＜0.05,P＜0.01),정일정제량의뢰성；형태상가견세포표면융모감소,내부공포상결구증가。소서류과민시험현시,2충약물대동물이람염솔균위100%、이문사람삼출량명현증다(P＜0.01)。리체여정체실험결과일치。결론청개령、혈새통주사액구유잠재적치류과민반응성,RBL-2H3세포모형용우중약주사제류과민성평개구유일정적삼고응용개치。
Objective To study anaphylactoid reactions induced by traditional Chinese medicine injections (TCMI) of Qingkailing (QKL) and Xuesaitong (XST) with RBL-2H3 cells;To provide some reference for improving the screening system of TCMI induced anaphylactoid reactions.Methods The IC50 induced by QKL and XST injections was determined by MTT assay. RBL-2H3 cells were stimulated with TCMI at different concentrations or with C48/80 or culture medium. Thirty minutes later, the supernatant was collected to determine the release of histamine andβ-hexosaminidase. The cell degranulation rate and the ultrastructure changes were observed. The ICR mice were given single injection of TCMI containing Evans Blue through tail vein. The number of the animal with blue ear, the total number of blue ears and the quantity of Evans Blue of extravasation were determined 30 minutes later.Results The IC50 of both QKL injection and XST injection was 12.5μL/mL. These two injections promoted RBL-2H3 cells to release histamine andβ-hexosaminidase at higher concentrations (P<0.05,P<0.01) in a dose dependent manner. Cell morphology showed a decrease of villous on the cell surface and an increase of the internal vacuolated structure. Both injections caused the blue ears of all animal with a rate of 100%. The quantity of Evans Blue of the extravasation was significantly increased (P<0.01). The results in vitro study were in close agreement with that in vivo.Conclusion Both QKL injection and XST injection may potentially cause anaphylactoid reactions. The RBL-2H3 cell model may be valuable to evaluate the anaphylactoid reactions induced by TCMI.