基于分子对接技术的注射用双黄连中有效成分致类过敏性实验研究  

Experimental study on the allergenicity of active ingredients in injectable Shuanghuanglian based on molecular docking technology

作　　者：曹建生 冯宇飞[1] 鲁文飞 吕颖 李文晴 廖威 CAO Jian-sheng;FENG Yu-fei;LU Wen-fei;LU Ying;LI Wen-qing;LIAO Wei(Heilongjiang University of Chinese Medicine,Harbin,Heilongjiang 150040,China)

机构地区：[1]黑龙江中医药大学,北药基础与应用研究教育部重点实验室,黑龙江哈尔滨150040

出　　处：《时珍国医国药》2024年第8期1894-1898,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(82174232);黑龙江省自然科学基金优秀青年项目(YQ2019H031)。

摘　　要：目的 筛选注射用双黄连中存在的致类过敏成分并对作用机制进行初步探讨。方法 以人类相关G蛋白偶联受体X2(Mas-related G protein-coupled receptor X2,MRGPRX2)为目标蛋白,通过AutoDock和Pymol软件与注射用双黄连中的活性物质作为配体进行分子对接,初步筛选致类过敏成分,并对RBL-2H3细胞的最佳实验条件(IC50)进行考察,将注射用双黄连活性成分中初选的成分与RBL-2H3细胞进行孵育,通过双抗夹心ELISA法与底物法检测组胺和β-氨基己糖苷酶的释放率,进一步筛分致类过敏成分,并观察其对RBL-2H3细胞内补体水平和细胞脱颗粒程度的影响,最后通过大鼠离体回肠实验进行验证。结果 RBL-2H3细胞分别给予注射用双黄连(28.25 mg/L)、β-谷甾醇(0.95 mg/L)、β-胡萝卜素(4.10 mg/L)、黄芩苷(1.90 mg/L)、咖啡酸甲酯(3.20 mg/L)、阿魏酸(2.29 mg/L)、绿原酸(1.51 mg/L)作用后,His、β-Hex释放均显著升高(P<0.01、0.05)。给药后的鼠总补体水平CH50和C3a、C5a水平均显著升高(P<0.05),并伴有脱颗粒现象。经离体回肠实验后β-谷甾醇、β-胡萝卜素、黄芩苷、咖啡酸甲酯、阿魏酸、绿原酸均能使大鼠回肠张力增加(P<0.01、0.05)。结论 注射用双黄连诱导的类过敏反应可能与β-谷甾醇、β-胡萝卜素、黄芩苷、咖啡酸甲酯、阿魏酸、绿原酸有关,且诱导的类过敏反应的作用机制可能与补体水平相关。Objective Screening for the presence of allergenic components in Shuanghuanglian injection and a preliminary inves-tigation of the mechanism of action.Methods With the target protein of Mas-related G protein-coupled receptor X2(MRC-PRX2)as the target protein,molecular docking with the active substance in injectable Shuanghuanglian as the ligand was carried out by AutoDock and Pymol sofware to preliminarily screen for class allergenic components,and the optimal RBL-2H3 cell ex-perimental conditions(IC50)were examined,the initially selected components of the active ingredients of Shuanghuanglian injec-tion were incubated with RBL-2H3 cells,and the release rate of histamine andβ-aminohexosidase was detected by double-antibody sandwich ELISA with substrate assay,to further screen the class-causing periplasmic components,and to observe their effects on the intracellular complement level and the degree of cell degranulation in RBL-2H3 cells,which were finally verified by a rat isolated ileum experiment for verification.Results His andβ-Hex release were significantly elevated in RBL-2H3 cells after administration of Shuanghuanglian injection(28.25 mg/L),bate-sitosterol,(0.95 mg/L),bate-carotene(4.10 mg/L),baicalein(1.90 mg/L),methyl caffeate(3.20 mg/L),ferulic acid(2.29 mg/L)and chlorogenic acid(1.51 mg/L),respectively(P<0.01,0.05).Total complement level CH50 and C3a and C5a levels were significantly elevated(P<0.05)with degranulation in rats after administration.After isolated ileum testβ-sitosterol,β-carotene,baicalein,methyl caffeate,ferulic acid and chlorogenic acid increased ileal tone in rats(P<0.01,0.05).Conclusion The anaphylactoid reaction induced by Shuanghuanglian injection may be related toβ-sitosterol,β-carotene,baicalein,methyl caffeate,ferulic acid and chloro-genic acid,and the mechanism of action of the induced anaphylactoid reaction may be related to the level of complement.

关 键 词：注射用双黄连 类过敏反应 分子对接 RBL-2H3细胞 Β-谷甾醇 Β-胡萝卜素 黄芩苷 咖啡酸甲酯 阿魏酸 绿原酸 

分 类 号：R283[医药卫生—中药学] R284.2[医药卫生—中医学]