| Objective: To study the expression of IL-25 and mast cell chymase in rats during anaphylactic shock.Methods:Male rats(SD)were randomly divided into normal control group and experimental group.The experimental group was injected intraperitoneally with chicken egg protein on day 1 and 5 days.At the 18 th day after the first administration the egg protein was stimulated by intravenous injection in tail to stimute anaphylactic shock.At the same time normal control group was injected with physiological saline.After the model was completed,the experimental group was divided into experimental control group and experimental death group.The experimental control group was the rats that did not die of anaphylactic shock and the experimental death group was the rats that did die of anaphylactic shock: postmortem normal temperature for 0h,12 h,refrigerated temperature for 12 h,24h,48 h,72h(6 rats in each group).At the end,the levels of Ig E,IL-25 and mast cell chymase in serum were respectively measured by enzyme-linked immunosorbent assay(ELISA).At the same time some organs(larynx,trachea,lung,heart,liver,kidney and small intestine)were drawn at the specified time and then placed in 10% neutral formaldehyde for 48 h,which was dehydrated,impregnated,embedded and made of wax block.And then the HE staining through the slice observe the pathological changes in the organs;The target protein from the lung,heart and small intestine sections was stained by immunohistochemical staining.The expression of IL-25 and chymase were observed under microscope and the images were collected.Again with the aid of computer image processing technology to analyze IOD(integral optical density)of IL-25 and chymase.When we analyze experimental data,the changes of the two indexes in the different storage periods of the anaphylactic shock were compared with those of the normal control group so as to judge the value of these two indexes in in the area of anaphylactic shock.Results: 1)Serum IgE,IL-25 and chymase analysis results:(1)The levels of IgE in normal control group and experimental control group(not die of anaphylactic shock group),anaphylactic shock death at normal temperature for 0h,normal temperature for12 h,refrigerated temperature for 12 h,24h,48 h,72h were compared P <0.01;The levels of IL-25 and chymase were compared by the above method,and the same was obtained.(2)The levels of Ig E in experimental control group and anaphylactic shock death in each group were compared P <0.01;The levels of IL-25 and chymase were compared by the above method,and the same was obtained.(3)The levels of Ig E in anaphylactic shock death at normal temperature for 0h and other shock death groups were compared P﹥0.05;The levels of IL-25 and chymase were compared by the above method,and the same was obtained;The data found that serum Ig E levels in the normal temperature 0h group was the largest,the other group was decreased,the serum IL-25 levels is also similar;serum chymase levels in the normal temperature 12 h group was the largest,the other group was decreased.2)IL-25 analysis results:(1)In lung tissues,IL-25 IOD value of normal control group and experimental control group,anaphylactic shock death in each group were compared P <0.05;IOD value of experimental control group and anaphylactic shock death at normal temperature for 0h were compared P <0.05,and anaphylactic shock death at normal temperature for12 h,refrigerated temperature for 12 h,24h,48 h,72h were compared P﹥0.05;IOD value of anaphylactic shock death at normal temperature for 0h and refrigerated temperature for 48 h,72h were compared P <0.05,and anaphylactic shock death at normal temperature for 12 h,refrigerated temperature for 12 h,24h were compared P ﹥0.05;IOD value of anaphylactic shock death at normal temperature for 0h was largest and other groups are reduced in turn;(2)In the myocardium,IL-25 IOD value of normal control group and experimental control group,anaphylactic shock death in each group were compared P <0.05;IOD value of experimental control group and anaphylactic shock death at normal temperature for 0h,refrigerated temperature for 12 h,24h,48 h,72h were compared P <0.05,and anaphylactic shock death at normal temperature for 12 h were compared P ﹥ 0.05;IOD value of anaphylactic shock death at normal temperature for 0h and other shock death groups were compared P﹥0.05;IOD average value of anaphylactic shock death at normal temperature for 0h was largest and other groups are reduced in turn,degree of declining at normal temperature for 12 h was more than refrigerated temperature for 12h;(3)In the small intestine,IL-25 IOD value of normal control group and experimental control group,anaphylactic shock death in each group were compared P<0.05;IOD value of experimental control group and anaphylactic shock death in each group were compared P ﹥ 0.05;IOD value of anaphylactic shock death at normal temperature for 0h and other shock death groups were compared P﹥0.05;IOD value of anaphylactic shock death at normal temperature for 0h was largest and other groups are reduced.3)chymase analysis results:(1)In lung tissues,chymase IOD value of normal control group and experimental control group,anaphylactic shock death in each group were compared P <0.05;IOD value of experimental control group and anaphylactic shock death at normal temperature for0 h,refrigerated temperature for 12 h,24h were compared P <0.05,and anaphylactic shock death at normal temperature for 12 h,refrigerated temperature for 48 h,72h were compared P﹥0.05;IOD value of anaphylactic shock death at normal temperature for 0h and normal temperature for 12 h,refrigerated temperature for 48 h,72h were compared P<0.05,and anaphylactic shock death at refrigerated temperature for 12 h,24h were compared P ﹥ 0.05;IOD everage value of anaphylactic shock death at normal temperature for 0h was largest and other groups are reduced in turn;(2)In the myocardium,chymase IOD value of normal control group and experimental control group,anaphylactic shock death in each group were compared P <0.05;IOD value of experimental control group and anaphylactic shock death in each group were compared P <0.05;IOD value of anaphylactic shock death at normal temperature for 0h and other shock death groups were compared P﹥0.05;IOD average value of anaphylactic shock death at normal temperature for 0h was largest and other groups are reduced and degree of declining at normal temperature for 12 h was more than refrigerated temperature for12h;(3)In the small intestine,chymase IOD value of normal control group and experimental control group,anaphylactic shock death in each group were compared P<0.05;IOD value of experimental control group and anaphylactic shock death in each group were compared P <0.05;IOD value of anaphylactic shock death at normal temperature for 0h and other shock death groups were compared P﹥0.05;IOD value of anaphylactic shock death at normal temperature for 0h was largest and other groups are reduced.Conclusion: 1)The expression of serum IgE in anaphylactic shock rats was significantly higher and it proved that the model is successful;serum Ig E levels and shock severity was positively correlated.2)In serum,lung,myocardium and small intestine of rats with anaphylactic shock,the expression of IL-25 had been increased and with the extension of the post-mortem preservation time the expression showed a downward trend.The level of IL-25 in serum was positively correlated with the severity of shock.3)In serum,lung,myocardium and small intestine of rats with anaphylactic shock,the expression of chymase had been increased and with the extension of the post-mortem preservation time the expression showed a downward trend.The level of chymase in serum,myocardium and small intestine organs,was positively correlated with the severity of shock. |
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