RBL-2H3细胞脱颗粒模型评价注射用丹参多酚酸的安全性

Evaluate safety of Salvianolic Acid for Injection with RBL-2H3 cell degranulation model

目的通过RBL-2H3肥大细胞脱颗粒模型考察注射用丹参多酚酸及丹参多酚酸B、D的致敏性。方法RBL-2H3肥大细胞分为对照(PBS)组、C48/80(阳性对照,4 mg/mL)组和梯度浓度(0.025、0.050、0.100、0.200、0.400、0.800 mg/mL)的注射用丹参多酚酸、丹参多酚酸B、丹参多酚酸D组,孵育30 min后,通过中性红染色试验观察脱颗粒现象,化学荧光法测定组胺释放率,酶联免疫吸附(ELISA)法测定β-氨基己糖苷酶释放率、类胰蛋白酶释放量。结果与对照组比较,阳性药C48/80组、质量浓度≥0.2mg/mL的丹参多酚酸B组、质量浓度≥0.4mg/mL的丹参多酚酸D组、质量浓度为0.8mg/mL的注射用丹参多酚酸组的细胞脱颗粒度显著升高(P<0.01、0.05);注射用丹参多酚酸没有引起RBL-2H3细胞组胺释放率增加,C48/80组、质量浓度≥0.025mg/mL的丹参多酚酸D组、0.8mg/mL的丹参多酚酸B组组胺释放率即显著升高(P<0.001、0.05);注射用丹参多酚酸和丹参多酚酸B组类胰蛋白酶释放量无显著性差异,质量浓度≥0.025 mg/mL的丹参多酚酸D组类胰蛋白酶释放量显著增加(P<0.001);注射用丹参多酚酸没有引起β-氨基己糖苷酶释放率增加,0.8mg/mL的丹参多酚酸D组、质量浓度≥0.1mg/mL的丹参多酚酸B组的β-氨基己糖苷酶释放率显著增加(P<0.001)。结论注射用丹参多酚酸在致敏性方面有较好的安全性。

Objective To study the sensitization of salvianolic acid B,D and Salvianolic Acid for Injection by RBL-2H3 mast cell degranulation model.Methods RBL-2H3 mast cells were divided into control group(PBS),C48/80(positive control,4 mg/mL)group and salvianolic acid,salvianolic acid B and salvianolic acid D groups at gradient concentrations(0.025,0.050,0.100,0.200,0.400,0.800 mg/mL)for injection.After incubation for 30 min,degranulation was observed by neutral red staining test.The release rate of histamine was determined by chemofluorescence method,and the release rate ofβ-hexaminosidase was determined by enzyme linked immunosorbent assay(ELISA).Results Compared with control group,the cell degranulation degree of positive drug C48/80 group,salvianolic acid B group with mass concentration≥0.2 mg/mL,salvianolic acid D group with mass concentration≥0.4 mg/mL and Salvianolic Acid for Injection group of 0.8 mg/mL were significantly increased(P<0.01,0.05).The histamine release rate of RBL-2H3 cells was not increased by Salvianolic Acid for Injection.The histamine release rate of C48/80 group,salvianolic acid D group with mass concentration≥0.025 mg/mL and salvianolic acid B group of 0.8 mg/mL were significantly increased(P<0.001,0.05).There was no significant difference in trypsinoid release between Salvianolic Acid for Injection and salvianolic acid B group,but the trypsinoid release of salvianolic acid D group with mass concentration≥0.025 mg/mL was significantly increased(P<0.001).The release rate ofβ-hexaminosidase was not increased by Salvianolic Acid for Injection,but significantly increased by 0.8 mg/mL salvianolic acid D and with mass concentration≥0.1 mg/mL salvianolic acid B(P<0.001).Conclusion Salvianolic Acid for Injection has good safety.