| Objective:To observe the changes of histamine and NO in anaphylactic shock model of rats with the passage of time, and explore Eotaxin and its contribution to anaphylactic shock, trying to seek objective morphological reference support to diagnose sudden erethistic death for forensic medicine.Methods:(1) Rats were divided into two groups randomly, the experimental group and the control group. Rats in experimental group were made into anaphylacatic shock modes with OVA. Lungs were extracted in different shock interval, including before sudden shock, 0.5 hours after shock, 12 hours after shock, 24 hours after shock,48 hours after shock.(2) To improve shore's fluorescence method to determine the level of histamine with time (3)To improve nitrate disoxidation method to determine the content of NO with time.(4) It was confirmed that NO and vasopermeability had a direct relation by experiment.(5)To apply enzyme chemical staining technique to stain NOS (6) Eotaxin was stained by using immunohistochemistry method, and the images of Eotaxin were analyzed by BI-2000, and the integral optical density (IOD) were calculatedResults: (1) The level of histamine in the sensitized group before exciting was significantly higher than the normal control group (P<0.01).It immediately and rapidly declined after allergens stimulated, lower than the control group, only 40 percent of the pre-excitation, the difference was significant. After a gradually rising trend, and that is, after 12 hours was higher than the normal control group, after 48 hours returned to 95% of the level of prior to stimulating, close to basic level before stimulating. And the level of histamine in normal control group were not obviously changed. (2) The content of NO in the sensitized group before exciting was significantly higher than the normal control group (P<0.01).It immediately and rapidly declined after allergens stimulated, lower than the control group, only 49 percent of the pre-excitation, the difference was significant. After a gradually rising trend, and that is, after 12 hours was higher than the normal control group, after 48 hours returned to 96% of the level of prior to stimulating, close to basic level before stimulating. And the content of NO in normal control group were not obviously changed. (3) Enzyme histochemistry results: NOS in the sensitized group was positive. It suggested as anaphylactic shock happened, the activity of NOS grew strengthened. (4) IHC results: the protein expression of Eotaxin of the experimental group were higher than that of the control group.(P<0.01)Conclusion:As anaphylactic shock happened , the level of histamine and the content of NO had a dynamic change in the pathogenesis of allergic shock. After statistical analysis,the level of histamine and the content of NO were positively correlated, and the relevance was very significant. It was confirmed that NO and vasopermeability had a direct relation by experiment.These supported that the application of enzyme histochemistry on staining NOS provided an objective morphological reference support for diagnosing sudden erethistic death in forensic medicine. And the protein expression of Eotaxin was significantly stronger in lung tissue of rat anaphylactic shock.so,the application of enzyme histochemistry on staining NOS and immunohistochemical technique on staining Eotaxin provided objective morphological reference supports for diagnosing sudden erethistic death in forensic medicine. |
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