Study On Sensitization Characteristics And Sensitization Markers Of Shuanghuanglian Injection

TraditionalChinese medicine injection (TCMI) is a perfect combination of tradi-tional Chinese medicine and modern pharmaceutical technology. There are more than 400 companies has been allowed to product 109 kinds of traditional Chinese medicine injections for more than 400 million users and creat more than 20 billion yuanannual sales. But in recent years, with extensive use of TCMI a series of adverse reactions (ADR)due to its composition and complexity manufacturing process is coming. Shuanghuanglian injection (SHLI)composed with extractions of honeysuckle, Scutel-laria and Forsythia.SHLI has the efficacy of detoxification and antiviral activity. SHLI have a good performance on the treatment of respiratory tract infections and other in-flammations in clinic.But with the increase use of SHLI is ADR gradually re-vealed.since SHLI appeared on the market in 2002, it has been token control measures twice due to the occurrence of severe ADR in the first issue of November 2001 andthe irst 22th issuein May 2009 of "adverse Drug reaction information Bulletin" national adverse by Drug reaction monitoring Center. SHLI is a large variety of TCMI, is ranked in top of TCMI ADRranking.Be a Case in Study of TCMI ADR have repre-sentative and practical significance. In this work, its characteristics of allergy in vivo, in vitro and markers of anaphylactoid reaction were researched.1. Allergic characteristics of SHLI in vivo.ObjectivesTo determine the type of SHLI allergy. Study the characteristics in vivo. MethodsFemale Balb/c mice, weighted in 18～20 g. Accordingto the method of ac-tive systemic anaphylaxis (ASA) establish SHLI type I allergy model. Set control group, positive drug group (OVA) and SHLI clinical dose groupto study allergic symptoms and permeabilitafter changesin mouse skin after drug challenge. According to clinical administration (one-time, intravenous) of SHLI establish anaphylactoid model.Set up of five graded dose of SHLI, SHLI5,10,20,40,60g · kg-1 (equivalent to clinical doses of 0.5,1,2x,4x and 6x). Study anaphylactoid reaction symptoms, IgE and histamine contentschange in serum, permeability changes in the intestinalwall, mesenteric vascular and skin after administered inlh.ResuItsBalb/c mice with SHLI clinical dose did not performenced anaphylaxis reaction inType I allergy mold ac-cording to the allergic symptoms and permeabilitafter changesin mouse skin after drug challenge. Inanaphylactoid reaction model, Balb/c mice with high doses SHLI (SHLI40,60g · kg-1) showed difficulty in breathing, edema in skin and eyes, eye se-cretions increased and other serious allergy symptoms in a short time after adminis-trated. Compared with control group the permeability of intestinal wall, mesenteric vascular and skin is increasesd. Serum IgE levels in mice of mice in each group did not change significantly, compared with negative group mice with drug the serum histamine content increased significantly. Conclusion Common clinical allergy symptoms is anaphylactoid reaction, its can occurs in a short timeafter administrated with drug.2. Research the anaphylactoid reaction markers in body of mice.ObjectiveSelect the proteins who responsedand worked in the anaphylactoid re-actionof SHLI in body of mice. MethodsPrepared serum samples with the method of chapter2.Cluster analysis protein in sample of each treatment group and the control group. Use TMT chemical labeling method andQ-Exative solution MS platform to detect the change of proteins in samples. ResultsCluster analysis showed that the protein from one grop serum with similar protein, clustering clear and easy to distin-guish. It was found 98 suspicious protiens that changed in the relative value of the protein content> 1.5 or<0.67 and P<0.05.Conclusion Exist some proteins who re-sponsedand worked in the anaphylactoid reactionof SHLI in body of mice. This work prepared some base work for for the screening of specific proteins.3. Anaphylactoid reaction characteristics of SHLI in vitro.ObjectiveObservethe influence of different concentration of SHLI and its inter-mediates on proliferation of RBL - 2H3 cells. Observethe influence of different con-centration of SHLI on apototisis and degranulation. Methods RBL-2H3 cells were treated with different concentration of SHLI (2 times gradient diluted from volume ratio 1:16 to 1:4096 and its intermediates (2 times gradient diluted from volume ratio 1:8 to 1:2048).The cellular morphology was observed under inverted microscope. MTT colorimetric assay was used to determine the growth of RBL-2H3 cells in dif-ferent concentration of SHLI and its intermediates, and the half inhibitory concentra-tion (IC50) was calculated. Dependent on the value of IC50, RBL-2H3 cells were then treated with different concentrations of SHLI to observe the effect of SHLI on cell apoptosis anddegranulation. Results As the concentration increases of SHLI and its intermediates, there inhibitory effect on proliferation of RBL-2H3 cells was gradually increased; the cells survival rate was decreased significantly;cells injury was gradual-ly aggravated; and the celluar morphology was changed, calculating their IC50. High dose SHLI could induce apotosis of RBL-2H3 cell. ConclusionSHLIand its interme-diates in high conclution could influence the proliferation of RBL-2H3 cells.SHLI in high conclution could not cause RBL-2H3 cells degranulation directly. This work can provide certain experiment basis for establishing anaphylactoid cell models and mon-itoring drug quality and safety.