The Anaphylactoid Constituents in Xue-Sai-Tong Injection

Abstract

Xue-Sai-Tong injection, a traditional Chinese medicine with total saponins of Sanqi ginseng as active ingredients, has been used for more than 500 years to treat coronary artery disease in China. Anaphylactoid reaction induced by Xue-Sai-Tong injection was one of the main adverse drug reactions which has occurred frequently in recent years. It is of importance to elucidate its anaphylactoid constituents. The in vivo anaphyalctoid tests indicated that the anaphylactoid mediators could be used as indexes to evaluate the anaphylactoid action. Further, the in vitro model based on determining the mediators release from the degranulation of mast cells and RBL-2H3 cells stimulated by Xue-Sai-Tong injection was explored. Mediators released from mast cells and RBL-2H3 cells caused by Xue-Sai-Tong injection were determined by comparison of the methods of fluorospectrophotometry, ELISA, and spectrophotometry, respectively, revealing that the histamine release induced by the Xue-Sai-Tong injection could not be assayed accurately by the method of fluorospectrophotometry because of the interference of saponins and unknown components in the injection. The rat peritoneal mast cell was also not an optimal cell model for determining histamine and β-hexosaminidase release due to the higher spontaneous release ratio during the cell collection. Thus, ELISA determination of the histamine release from RBL-2H3 cells is a suitable in vitro model to assay the anaphylactoid reaction of Xue-Sai-Tong injection. Previously, abnormal hemolysis in some batches of Xue-Sai-Tong injection was observed in the course of their HD₅₀ (half hemolytic dosage) determination. This study further found that injections which exhibited an abnormal hemolysis phenomenon also caused a higher release of the anaphylactoid mediators from RBL-2H3 cells, indicating the HD₅₀ could be an auxiliary index to evaluate anaphylactoid action of the herbal injection indirectly. Research for anaphylactoid components in Xue-Sai-Tong injection indicated that proteins with over 10 KDa of molecular weight, but not ginsenosides, could be the main constituents inducing the release of anaphylactoid mediators from RBL-2H3 cells. An HPLC method for protein determination in the Xue-Sai-Tong injection was established subsequently, and the content of proteins with molecular weights of over 10 KDa in the injections showed an obviously positive correlation with the histamine release induced by the injections. In addition, taking ginsenoside-Rd coupled with BSA as an example, the hapten property of ginsenosides was studied and the ratio of ginsenoside-Rd to BSA was determined to be 8 : 1 by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the ginsenoside-BSA conjugate showed a stronger action to stimulate histamine release from the RBL-2H3 cells.

Key words

herbal medicine injection - anaphylactoid reaction - Panax notoginseng - ginsenoside - RBL-2H3 cells - histamine - Araliaceae

• References

• 1 Xu XH. Severe adverse effects of Puerarin injection in 18 cases. SH J TCM 2006; 40: 71-72
• 2 Yi D, He Y, Zhou YL. Analysis of 26 adverse drug reactions cases of Ginkgo leaf injection. Chin J Pharm 2009; 6: 411-414
• 3 SFDA. Notice for the usage and registration of Yu-Xing-Cao etc. 7 kinds of herbal injections. Beijing: SFDA; 2006
• 4 Wang N, Wan JB, Li MY, Wang YT. Advances in studies on Panax notoginseng against atherosclerosis. Zhong Cao Yao 2008; 5: 787-791
• 5 Dou DQ, Zhang YW, Zhang L, Chen YJ, Yao XS. The inhibitory effects of ginsenosides on protein tyrosine activation induced by hypoxia/reoxygenation in cultured human umbilical vein endothelial cells. Planta Med 2001; 67: 19-23
• 6 Yao XH, Li XJ. Panax notoginseng saponins injection in treatment of cerebral infarction with a multicenter study. Chin J New Drugs Clin Rem 2001; 20: 257-260
• 7 Yu JL, Dou DQ, Chen XH, Yang HZ, Guo N, Cheng GF. Protopanaxatriol-type ginsenosides differentially modulate type 1 and type 2 cytokines production from murine splenocytes. Planta Med 2005; 71: 202-207
• 8 Qiu YK, Dou DQ, Cai LP, Jiang HP, Kang TG, Yang BY, Kuang HX, Li MZ. Dammarane-type saponins from Panax quinquefolium and their inhibition activity on human breast cancer MCF-7 cells. Fitoterapia 2009; 80: 219-222
• 9 De-Qiang D, Zheng X, Guang Y, Jian-Feng Z, Ying-Kun Q, Jing-Xian Y, Ting-Guo K. Prediagnostic methods for the hemolysis of herbal medicine injection. J Ethnopharmacol 2011; 138: 445-450
• 10 Han F, Zhou QA, Zhan M. Safety analysis of traditional Chinese medicine injection. Prog Vet Med 2007; 28: 104-107
• 11 Dou DQ, Chen YJ, Liang LH, Pang FG, Shimizu N, Takeda T. Six new dammarane-type triterpene saponins from the leaves of Panax ginseng . Chem Pharm Bull 2001; 49: 442-446
• 12 Jiang HP, Qiu YK, Cheng DR, Kang TG, Dou DQ. Structure elucidation and complete NMR spectral assignments of two new dammarane-type tetraglycosides from Panax quinquefolium . Magn Reson Chem 2008; 46: 786-790
• 13 Passante E, Ehrhardt C, Sheridan H, Frankish N. RBL-2H3 cells are an imprecise model for mast cell mediator release. Inflamm Res 2009; 58: 611-618
• 14 Yukihiro S, Hiroyuki T, Noriko F. Monoclonal antibodies against naturally occurring bioactive compounds. Cytotechnology 1999; 31: 9-27
• 15 Sun DR, Qi P. Adverse drug reaction. Beijing: Peopleʼs Medical Publishing House; 2003: 86
• 16 Coutts SM, Nehring RE, Jariwala NU. Purification of rat peritoneal mast cells: occupation of IgE-receptors by IgE prevents loss of the receptors. J Immunol 1980; 124: 2309-2315
• 17 Horigome K, Bullock ED, Johnson EM. Effects of nerve growth factor on rat peritoneal mast cells. Survival promotion and immediate-early gene induction. J Biol Chem 1994; 269: 2695-2702
• 18 Saito H, Kato A, Matsumoto K, Okayama Y. Culture of human mast cells from peripheral blood progenitors. Nat Protoc 2006; 1: 2178-2183