An Experimental Study Of Xiaomin Granules Antagonizing Type â…  Hypersensitivity

ObjectiveTo investigate the effect and mechanism of Xiaomin Granules on typeⅠhypersensitivity of cutaneous and systemic, provide theoretic evidence for the clinical application of Xiaomin Granules.MethodsThis research include three parts: First, to investigate the effect of Xiaomin Granules on cutaneous typeⅠhypersensitivity, this part include capillary permeability increasing test caused by histamine and 5-HT in the mice, Passive Cutaneous Anaphylaxis (PCA) test and footpad swelling test caused by histamine and 5-HT to investigate the effect of Xiaomin Granules to the increase of capillary permeability. Second, to investigate the effects of Xiaomin Granules on systemic typeⅠhypersensitivity, the model of anaphylactic shock of the mice caused by compound 48/80 and anaphylactic shock of the guinea-pig caused by antigen were used to investigate the effects of Xiaomin Granules on systemic typeⅠhypersensitivity. Third, to discuss the mechanism of Xiaomin Granules antagonizing typeⅠhypersensitivity, such as the effect of Xiaomin Granules on the mast cells degranulation in abdominal cavity, fluorometric assay was used to measure histamine released from mast cells of the rats, immunohistochemical method was used to investigate the effect on 5-HT released from mast cells, ELISA was used to measure the content of IgE in anaphylactic rats'serum and the content of cytokine IFN-γ,IL-4 in anaphylactic rats'serum, RT-PCR wad used to investigate the expression of IFN-γmRNA,IL-4mRNA in anaphylactic rats'spleen lymphocytes, flow cytometry and transmission electron microscope was used to observe the effect of Xiaomin Granules on T lymphocytes apoptosis.Results1. The effect of Xiaomin Granules in restraining permeability of capillary caused by histamine : the result indicated that Xiaomin Granules can inhibit the increasing of capillary permeability caused by histamine. There are remarkable differences between all dose groups and normal group of Xiaomin Granules.(low-dose group P<0.05,medium,high-groups P<0.01),the low,medium,high-dose group have obvious dose-effect relationship.2. The effect of Xiaomin Granules in capillary permeability of the mice caused by 5-HT: the result indicated that Xiaomin Granules can inhibit permeability of capillaries of the mice caused by 5-HT ,there is remarkable differences between all dose groups and normal group (P<0.01).3. The effect of Xiaomin Granules on Passive Cutaneous Anaphylaxis ears of the mice : the result indicated that PCA of all Xiaomin Granules groups have obviously reduced, compared with normal group, there is significant difference(P<0.01).4. The effect of Xiaomin Granules on mice footpad swelling caused by histamine: the result indicated that after histamine was injected 30min , low-dose group have certain inhibitory action, with the time extends, the function decreased. The medium-dose and high-dose groups may obviously inhibit the footpad swelling in whole time of this experiment.5. The effect of Xiaomin Granules on mice shock caused by Compound 48/80: the result indicated that Compound48/80 can induce the shock and death of the mice. On the contrary, Xiaomin Granules can decrease death rate of the mice, the effect of high-dose group is more better, compared with model group, there is difference(P=0.05).6. The effect of Xiaomin Granules on anaphylactic shock of the guinea-pigs caused by antigen: the result indicated that Xiaomin Granules can delay the sensitive time, compared with model group, there is significant difference(P<0.01).7. The effect of Xiaomin Granules on the secretion of IgE in the rats'serum : the result indicated that Xiaomin Granules can inhibit the secretion of IgE, there is significant difference between all groups of Xiaomin Granules and model group(P<0.01).8. The effect of Xiaomin Granules on degranulation of peritoneal mastocyte: the result indicated that degranulation of peritoneal mastocyte of all groups is much more than normal group , they have significant differences ,mast cells degranulation in abdominal cavity of all Xiaomin Granules groups is less than model group, they have significant differences(P<0.01).9. The effect of Xiaomin Granules on mast cells releasing histamine in abdominal cavity of the rats: the result indicated that the mount of mast cells releasing histamine in abdominal cavity of all Xiaomin Granules groups is much less than model group (P<0.01), high-dose Xiaomin Granule group and chlorpheniramine group is less than low-group, there are significant differences (P<0.01).10. The effect of Xiaomin Granules on peritoneal mastocyte releasing 5-hydroxy tryptamines of the rats: the result indicated that peritoneal mastocyte of all Xiaomin Granules groups have escaped pellets and the expression of 5- hydroxy tryptamines has weaken. All groups of Xiaomin Granules have significant differences compared with model group(P<0.01).11. The effect of Xiaomin Granules on the content of IFN-γand IL-4 in the rats'serum: the result indicated that the content of IFN-γin the rats'serum of Xiaomin Granules groups has increased, high-dose Xiaomin Granules group increased more obviously, there is significant difference (compared with model group, P<0.01). The content of IL-4 in the serum of all Xiaomin Granules groups has reduced, high-dose group of Xiaomin Granules reduced obviously, there is significant difference (compared with model group, P<0.01).The level of IFN-γ/ IL-4 in high-dose Xiaomin Granules group is higher than in model group, there is significant difference (P<0.01).12. The effect of Xiaomin Granules on the expression of IFN-γmRNA and IL-4mRNA in rats'spleen: the result of agarose electrophoresis indicated that the iuminous bandings can be obviously found, Xiaomin Granules group and chlorpheniramine group were more stronger than other groups. The iuminous bandings can be obviously found after IL-4 has amplified, its brightness stronger in the model group than in normal group and each medication group, the normal group banding is darkest. The result indicated that the expression of IFN-γmRNA in rats'spleen of high-dose Xiaomin Granules group has obviously increased and has remarkable difference compared with model group(P<0.01). The expression of IL-4 mRNA in rats'spleen is obviously higher in model group than in normal group(P<0.01),the expression of IL-4 mRNA in high-dose Xiaomin Granules group has obviously reduced(P<0.01).13. Using transmission electron microscope to observe the effects of Xiaomin Granules on apoptosis of spleen T lymphocytes : the result indicated that the performance of early time apoptosis of T lymphocytes and typical bodies of apoptosis are found in Xiaomin Granules group.14.Using transmission electron microscope to observe the effects of Xiaomin Granules on apoptosis of spleen T lymphocytes: the result indicated that the structure of spleen T lymphocytes of model group tend to be normal and has few apoptosis cells, necrosis spleen T lymphocyte may be found occasionally. Spleen T lymphocytes apoptosis occurs obviously in Xiaomin Granules group.15. The effect of Xiaomin Granules on apoptosis of T lymphocytes of peripheral blood determined by flow cytometer: the result indicated that the quantity of apoptosis of T lymphocytes of Xiaomin Granules group has obviously increased. There is significant difference between Xiaomin Granules group and model group in the percentage of apoptosis of T lymphocytes (P<0.01).Conclusions1. Xiaomin Granules has the function of inhibiting cutaneousⅠHypersensitivity, such as inhibiting the mice capillary permeability increase caused by histamine and 5-HT, inhibiting Passive Cutaneous Anaphylaxis ears of the mice, inhibiting mice footpad swelling caused by histamine.2. Xiaomin Granules has the function of inhibiting systemic typeⅠh ypersensitivity: reducing the death rate of shock in the mice caused by Compound 48/80, delay the sensitive time of guinea-pigs caused by antigen, high-dose group can reduce the death rate.3. Xiaomin Granules have many functions, such as reduce mast cells degranulation, reduce the release of histamine and 5-hydroxy tryptamine, reduce the content of IgE in the serum, increase the content of IFN-γin the serum, reduce the content of IL-4 in the serum, promote the expression of IFN-γmRNA, inhibit the expression of IL-4mRNA, up-regulate the Th1/Th2 proportion, induce the apoptosis of T lymphocytes and so on.Xiaomin Granules can antagonize typeⅠhypersensitivity through not only inhibiting the IgE production and reducing the release of biological activity medium from mast cells and other cells , but also by controlling the balance of Th1/Th2 , and inducing the apoptosis of T lymphocytes to perform the function of resistingⅠHypersensitivity.