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
