| Objective:According to the clinical application experience of Professor Xiang Nan for many years,Prunella vulgaris capsule has good curative effect on patients with autoimmune thyroiditis,and it's safe,but its mechanism of action is not yet clear.In this study,we estabilshed a pathological model of experimental autoimmune thyroiditis in rats.Selenium yeast tablets were used as a control group to observe the thyroid tissue of rats with autoimmune thyroiditis.Pathology,thyroidstimulating hormone,autoantibodies,Th cell-associated cytokines,expression of Fas/FasL protein,expression of regulatory T cell subsets,in-depth study of the immune regulation mechanism of Prunella vulgaris capsule on autoimmune thyroiditis in rats And the pathogenesis of the disease provides sufficient theoretical and experimental data for the clinical application of Prunella vulgaris capsules,and provides ideas for the development of new immunological preparations in the future.Methods:Forty 4-6 weeks SD female SD rats which weighed 100-120g were reared for 1 week and then randomly assigned to 4 groups according to the random number table method.:The normal group,model control group,selenium yeast group,Prunella vulgaris capsule group,10 in each group.Modeling:Porcine thyroglobulin(PTg)and complete Freund's adjuvant(CAF)were mixed and emulsified.Rats except normal controls were injected with PTg at multiple points in the back of the rat's plantar pad.The dose was 100?g/ml.Only for the first immunization,once a week,for 2 weeks,after 3 weeks,PTg and incomplete Freund's adjuvant(IAF)were mixed and emulsified,and the rats were injected subcutaneously at the inside and back of the limbs.strengthen 1 to the end of the 6th week and give sodium iodide water fed with high iodine.In the fourth week of the model began to be drug intervention,selenium yeast group was formulated into 20ug/ml selenium yeast solution,according to lml/lOOg.d each rat,Prunella capsules formulated into 50mg/ml Prunella vulgaris solution According to 1ml/100g.d,each rat was irrigated.The normal group and the model control group were given normal saline according to 1ml/100g.Each group of continuous drug intervention for 6 weeks.Rats were dynamically observed during the experiment.After the end of the experiment,rat serum,thyroid tissue,and spleen were taken.HE staining was used to observe the histopathological changes of the thyroid gland by light microscopy,and serum TSH was detected by enzyme-linked immunosorbent assay(ELISA).TGAb,TPOAb and Th related cytokines INF-?,TNF-?,IL-4,IL-17,IL-35 content,immunohistochemical method to observe the expression of Fas and FasL protein in rat thyroid tissue,flow method detection Percentage of CD4~+CD25~+Foxp3~+Treg cells in rat's spleen.Results:1.The titer of autoantibody TGAb and TPOAb in the serum of the model group was significantly higher than that in the blank control group(P<0.05).Obvious lymphocyte infiltration was seen in the thyroid tissue,which proved the successful manufacture of autoimmune thyroiditis.The mouse model laid the foundation for further experiments.2.The follicles of the thyroid group in the model group showed a large number of destruction under light microscope.The morphology was not regular,and a large number of lymphocyte infiltration was observed.After intervention of Selenium yeast and Prunella vulgaris,the follicular destruction and lymphocyte infiltration were compared with the model group.Improvement,and the improvement effect of Prunella vulgaris better than selenium yeast group.3.The levels of TSH,TGAb,TPOAb and Th-related cytokines IFN-?,TNF-?,IL-4 and IL-17 in the serum of the model group were significantly higher than those in the blank control group(P<0.05).The level of each index was significantly lower than that of the model group(P<0.05),and the Prunella vulgaris and selenium yeast group had relatively lower levels of TSH,IFN-?,IL-4 and IL-17than the selenium capsule group.In the yeast group(P<0.05),there was no difference in the level of TGAb between the selenium yeast group and the selenium yeast group(P>0.05).TPOAb and TNF-?levels were higher in the yeast selenium yeast group(P<0.05).4.The expression of Fas and FasL immunoreactive substances in the thyroid tissue of the model group was more than that of the model group.The color was deep and the range was wide.The expression score was higher than that of the blank control group(P<0.05).Fas and FasL expression was observed after drug intervention.Decreased,lighter color,narrower range,lower expression scores than the model group(P<0.05),and the expression level of immunopositives in the Prunella vulgaris capsule group was lower than that in the selenium yeast group(P<0.05).5.The ratio of CD4~+CD25~+Foxp3~+Treg in the spleen of rats in the model group was lower than that in the control group(P<0.05).The Prunella vulgaris capsule group and the selenium yeast group were higher than the model group(P<0.05).The Prunella vulgaris capsule group and selenium There was no statistical difference between the yeast groups(P>0.05).6.The serum IL-35 in the model group was lower than that in the blank control group(P<0.05),the Prunella vulgaris capsule group and the selenium yeast group were higher than the model group(P<0.05).There was no statistical comparison between the Prunella vulgaris capsule group and the selenium yeast group.Differences in learning(P>0.05).7.The level of IL-35 was positively correlated with the number of Treg in the spleen of rats(r=0.507,P=0.001<0.05).Conclusion:This experiment successfully produced a rat model of autoimmune thyroiditis and found that Th1/Th2,Th17-related pro-inflammatory cytokines were up-regulated and the number of Treg was decreased in AIT rats,and the Fas/FasL pathway was activated at the same time.The abnormal apoptosis of thyroid cells caused destruction of follicular cells and inflammatory infiltration,leading to increased levels of TSH,TGAb,and TPOAb.Xiecao Capsule can reduce the levels of Th-related cytokines IFN-?,TNF-?,IL-4,IL-17 in AIT model rats,increase the number of regulatory T cells and IL-35 levels,and reduce the thyroid tissue Fas,FasL protein The expression improves the destruction of follicles and lymphocyte infiltration in rat thyroid tissue,lowers the levels of TSH and autoantibodies TGAb and TPOAb,and exerts an immunomodulatory effect from both the inhibition of inflammatory activation and the promotion of inflammation inhibition. |
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