Preliminary Study On Sensitizing Substances Of Safflower Injection

ObjectivesThe paper preliminary revealed the material basis that may cause allergicreaction in safflower injection include macromolecule materials and small materials.The macromolecule materials were almost proteins directly stimulate body to provokean immune response, and then cause allergic reaction when it access to the humanbody. Small molecules such as oligosaccharide,pigment,polyphenols and fatty oil etc.They can be used as an antigen or hapten to cause allergic reaction.Methods1.Collect safflower injection that has clinical allergic reaction reports, and usingcavia procellus active systemic allergic reaction (ASA) and rat passive cutaneousanaphylaxis (PCA), then observe the allergy symptoms of cavia procellus anddetermine the diameter of rats’ back skin locus coeruleus, finally choose the samplesof safflower injection which can cause animal allergic reaction.2.Acetone precipitation method was used to initial separate protein material andsmall molecules in the allergic safflower injection, and two substances were tested forallergenic animal testing in order to intensive study the category of allergenic.3.Acetone precipitation method was used to obtain the crude protein, the proteinand was separated by Two-dimensional electrophoresis, Western-Blot method wereused to position the allergic proteins of safflower injection.4.The proteins were purified and enriched by ion exchange resin. Used ASA,PCA,and ELISA determine the content of allergic mediator histamine in serum, lung tissueand airway tissue homogenates, and lung tissue and airway tissue were biopsied.Multi-level co-verify the allergenicity of purified proteins.5.Small molecular substance was the solution portion which was same as4. Multi-levelco-verify the allergenicity of small molecular. Results1. Safflower injection of NO.07123101caused positive reaction of cavia procellus(the ratio was33.3%). the blue spot diameter of rat back skin was greater than5mm.This proved the sample has animal allergenicity.2.Protein substances obtained by acetone precipitation method had sensitizationthrough ASA, but further research will be needed.3.Crude proteins was separated by two-dimensional electrophoresis andsensitization by and Western Blot.Between40KD and60KD had two proteinpoints,about26KD has one protein points.4.Purified proteins caused positive reaction of BN rat, and the blue spot diameterof rat back skin was greater than5mm, the histamine content in serum and tissue hada significant difference with the negative (p<0.01), and the pathological section oflung tissue and airway tissue appeared different levels of inflammatory change.5.The lower content of small molecule in safflower injection did not cause allergicsymptoms of BN rats and rats, and the histamine content in serum and tissue had nota significant difference with the negative (p>0.05), the pathological section of lungtissue and airway tissue had not obvious pathological changes.ConclusionsThrough the preliminary study of experimental results, the material basis ofsafflower injection allergic reactions was mainly protein substances， its formulaweight between40and60KD,and about26KD. Small molecules cann’t make allergicreactions in rats,it was not main reson which caused allergic reaction.