Anaphylactoid Reactions Induced By Shuanghuanglian For Injecting And The Mechanism Study

The adverse drug reactions induced by Shuanghuanglian for Injecting(SHLI) occurred more and more frequently with its wide use in clinic,especially the hypersensitivity.SHLI is able to cause acute hypersensitive reaction(AHR),which includes anaphylactoid reaction and typeⅠallergy.The anaphylactoid reaction presents with an identical clinical picture of typeⅠallergy but without IgE involvement,occuring at the first expose to agents and more frequently.In our study, we proved that the AHRs induced by SHLI mainly include anaphylactoid reactions, and investigated the mechanism and system for detection,expecting to provide new ideas for establishment of pre-clinical evaluation methods of Traditional Chinese Medicine Injections.1 Study on the type of acute hypersensitive reactions induced by SHLIKM mice were injected with saline,Compound 48/80,SHLI,Baicalin, Chlorogenic acid(CGA) and Forsythin respectively,and signs of anaphylactoid reactions were observed.The results suggested that Compound 48/80 led to crawling, prostration and convulsion in mice,and SHLI at 360,720 and 1440 mg/kg caused staggering,crawling,prostration,convulsion and mortality,which were dose dependent.Mice dosed with Baicalin,CGA and Forsythin appeared normal.Rats were sensitized with saline,Ovalbumin and SHLI respectively for three times, and the blood samples were collected on day 10 after the last sensitization for IgE detection.The results showed that the total serum IgE concentration in Ovalbumin group was 147.9±25.5μg/mL,much higher than the normal level(p＜0.05),but the total serum IgE levels of SHLI groups were nearly equal to that of saline group.2 Study on the mechanism of anaphylactoid reactions induced by SHLIRat peritoneal mast cells(RPMC) were incubated with saline,Compound 48/80, SHLI,Baicalin,CGA and Forsythin respectively in vitro,and the histamine releases were examined.The results showed that the histamine release rate of Compound 48/80 group was 76.9±1.8%,higher than the saline group(p＜0.01),but no significant difference was observed between the SHLI,Baicalin,CGA,Forsythin group and the saline group.Rats were injected with saline,Compound 48/80 and SHLI respectively,and the levels of histamine and tryptase in the blood were measured.The results demonstrated that Compound 48/80 and SHLI both increased the histamine and tryptase level in vivo(p＜0.01).3 Investigation on detection system of anaphylactoid reactions induced by SHLIMean arterial pressure(MAP) and heart rate in response to different drugs were measured in anesthetized rats.3.1 Responses to SHLIFollowing one bolus injection of histamine at 0.1,1.0 and 5.0 mg/kg iv,MAP decreased immediately,and the maximum magnitude of decrease were 59.2,46.0 and 64.3 mmHg respectively.Increasing doses of histamine prolonged the duration of action.Intravenous bolus injection of SHLI resulted in immediate decrease in MAP at 100,200 and 400 mg/kg in anesthetized rats,with the maximum magnitude of decrease 89.1,84.8 and 90.1 mmHg respectively.About 2 to 3 min after the initiation of injection,the blood pressure returned to the basal level.The heart rate increased, decreased or kept still after administration of histamine or SHLI without obvious regulation.When histamine was given twice at 1.0 mg/kg,the hypotensive responses were similar.Repetitive dosing of SHLI demonstrated similar effect with histamine.3.2 Effect of Diphenhydramine on hypotensive responses induced by histamine and SHLIWhen rats were administered with histamine at 0.1 mg/kg twice with Diphenhydramine given before the second histamine injection,the mean changes of MAP from baseline to nadir were 69.1 mmHg and 15.3 mmHg after each histamine administration respectively(p＜0.01).When rats were administered with SHLI at 0.1 mg/kg twice with Diphenhydramine given before the second SHLI injection,the mean changes of MAP from baseline to nadir were 60.3 mmHg and 38.1 mmHg after each SHLI administration respectively(p＜0.01).3.3 Effect of Cromolyn on hypotensive responses induced by Compound 48/80 and SHLIWhen Compound 48/80 was used,MAP decreased significantly from 162.8±12.4 mmHg to the nadir of 104.5±12.0 mmHg(p＜0.003).When Compound 48/80 was given after the infusion of Cromolyn,the MAP did not change.When SHLI(200 mg/kg) was injected,MAP decreased rapidly from 156.7±4.6 mmHg to 81.4±6.9 mmHg(p＜0.00005).Pretreatment with Cromolyn could not attenuate the hypotensive responses induced by SHLI,77.6±4.6 mmHg vs.baseline 163.4±4.6 mmHg(p＜0.0002)3.4 Effect of infusion speed on hypotensive responses induced by SHLISHLI(400 mg/kg,4.0 mL) was infused to rats at the speed of 0.8,0.4 or 0.2 mL/min.The results showed that the maximum magnitude of decrease in MAP increased with infusion speed,and the initiation and ending of hypotension consisted with that of infusion.No hypotension was observed when the infusion speed decreased to 0.2 mL/min.4 Hypotensive materials detection SHLI was observed to induce hypotensive responses at 2.8 mg/kg in cats by using the hypotensive materials detection method described in the Chinese Pharmacopoeia.In conclusion,the hypersensitive reactions induced by SHLI may include anaphylactoid reactions primarily.The mechanism is possible to be mediators release from mast cells stimulated by SHLI,which leads to toxical signs of multiple systems, hypotention for instance.Histamine similar materials may be contained in SHLI, possible to contribute to the hypotension.With the merits of sensitivity and rapidity, hypertensive materials detection may act as a standard test for detection of anaphylactoid reactions induced by SHLI.It is recommended that the safety evaluation of SHLI should include hypertensive materials detection.