| Background and ObjectiveChemokines are low molecular weight polypeptide that divided into groups based on a cysteine motif, that is, CXC, CC,C, and CX3C, in which the C is a cysteine and X stands for any amino acid residue.To date, around50chemokines have been identified in humans. The recruitment of leukocytes, such as lymphocte, monocyte, neutrophil to inflammatory sites was. the first function attributed to chemokines, it mediate the immune cells infiltration, location and activation in organization, and they have chemotactic function of lymphocyte and forming lymphoid follicles, subsequently, chemokines play a role in autoimmune thyroid disorders.MIP-1α, MIP-1β and MCP-1are important chemokines in humun body, and they were proved the participation in occurrence and development of a wide variety of autoimmune diseases. Both GD and HT are the most common human autoimmune diseases. They are characterized of the production of antibodies and lymphocytic infiltration of the thyroid, which can result in tissue destruction. There were no research at home and less research abroad about the hemokines, MIP-1α, MIP-1β and MCP-1, whether or not participate in the development of autoimmune thyroid disease. This study applies SP immunohistochemical method and ELISA method to observe the expression of MIP-1alpha, beta and MIP-1in GD and HT, thyroid adenoma, nodular goiter and normal thyroid tissue and serum, and the correlation analysis between chemokines expression and serum thyroid autoantibodies is done, thus to explore these chemokines in its role in the development of autoimmune thyroid disease.Methods44cases of Graves disease (GD)(33male cases, female11cases), age (33.4±1.51),19cases of Hashimoto thyroiditis (HT)(male0case, female19cases), age (47.4±2.24),14cases of thyroid adenoma(male4cases, female10cases), age (42.3±4.07),10cases of multinodular goiter(male4cases, female6cases), age (55.1±3.32) and normal tissue(male2cases, female6cases), age (43.3±3.08), surrounding thyroid adenoma specimen as normal controls. Thyroid specimen and preoperative serum were collected from patients being hospitalized in our hospital from year2009to2011with thyroid surgery. Another8cases (male3cases, female5cases), age (40.3±1.08). Blood samples were drawed from medical check-up healthy subjects as normal controls. Inclusion criteria:all cases were confirmed by clinical and laboratory examination and pathological diagnosis. Exclusion criteria:recent history of infection, women’s pregnancy, tumors, lung disease, cardiovascular disease, organ transplant, infection with other autoimmune diseases. All the tissue samples were treated with HE and immunohistochemical staining after fixation, dehydration, embedding and sectioning. Each section selected10non-overlapping horizons at high magnification randomly. Results judgement:cytoplasm containing yellow particles for positive cells. Scoring Standards:The product of the staining intensity rating (not dyeing:0score, weak yellow,1score, blond:2score, brown:3score,) and cell positive score (cell positive rate<5%for0,5%～-25) for1score,>25%～50%for the2score,>50%and75%for the three score,> to4score) as the result of the case. According to the scores indicate expression intensity,0is classified at negative (-),1～2is classified at weak positive (+),3～6is classified at positive (++),>6for strong positive (+++). We determine serum specimens with double antibody concentration of specimens in three kinds of chemokines. Serum TRAb, TGAb and TPOAb are collected by preopreative examinnation in our hospital. All the data were counted and analyzed with SPSS17.0statistics software package and positive compare use nonparametric test of level data. Nonparametric test was used for quantitative data. Spearman rank correlation analysis were used for the correlation of Chemokines expression and serum TRAb,TGAb, TPOAb.. P<0.05(double side) was statistically significant,ResultsUsing SP immunohistochemical method to detect the expression MIP-1alpha, beta and MCP-1in44cases of Graves disease,19cases of hashimoto’s thyroiditis,14cases of thyroid adenoma,10cases of nodular goiter and8cases of normal thyroid tissue. The study showed that in the organization of Graves disease and hashimoto’s thyroiditis, the immune reaction product MIP-1alpha, beta and MIP-1positioned in thyroid follicular cells, it showed a strong exepression in thyroid follicular cells near by inflammatory cells. In TH organization, the nucleus of thyroid follicular epithelial cell and lymphoid cells of part lymphoid follicular germinal center also have positive expression. The residual thyroid follicular in the stroma cells infiltrating by a large number of lymphocytes and plasma cells almost all expressed. HE staining of eosinophilic cells express intensity higher, dyed brown crumbly. The expression in thyroid adenoma and nodular goiter were moderate expression, dyed yellow or pale yellow. Almost no expression in normal thyroid tissue, very little of epithelial cells in light yellow.The expression of MIP-1alpha, beta and MCP-1in Graves disease, hashimoto thyroiditis group are significantly higher than in normal thyroid tissues, nodular goiter and thyroid adenoma (P<0.05). There was no significant difference between groups of Graves disease and hashimoto thyroiditis (P>0.05), Also,there was no significant difference in groups of normal thyroid tissues, nodular goiter and thyroid adenoma.Spearman correlation analysis showed that the expression of MIP-1alpha in Graves disease is closely correlated to the distribution and the serum TRAb value (r= 0.504, p=0.365), and are closely correlated to the TGAb value (r=0.365, p=0.365), and closely correlated to the TPOAb value (r=0.561, p=0.561); The expression of MIP-1beta in Graves disease is closely correlated to an the serum TRAb value0=0.419, p=0.003), closely correlated to the TGAb value (r=0.434, p=0.003), also closely correlated to the TPOAb value0=0.491,p=0.001); The expression of MCP-1in Graves disease is closely correlated to the serum TRAb value (r=0.482, p=0.007), closely correlated to the TGAb value0=0.398,p=0.007), also closely correlated to the TPOAb value0=0.456, p=0.002). The expression of MIP-1alpha in hashimoto thyroiditis is closely correlated to the serum TGAb value (r=0.501, p=0.019), closely correlated to the TPOAb value (r=0.397,p=0.026); The expression of MIP-1beta in hashimoto thyroiditis is closely correlated to the serum TGAb value (r=0.536, p=0.018), and closely correlated to the TPOAb value (r=0.426, p=0.006); The expression of MCP-1in hashimoto thyroiditis is closely correlated to the serum TGAb value (r=0.051,p=0.019), also closely correlated to the TPOAb value (r=0.397,p=0.026)And serum MIP-1alpha, beta and MIP-1levels were detected in five groups using of ELISA. In serum levels of MIP-1alpha there was no statistically significant difference among the groups (X2=0.326, P>0.056); In serum levels of MIP-1beta, there was also no statistically significant difference among the groups (X2=2.600, P>0.05). In serum levels of MCP-1there was also no statistically significant difference among groups (X2=5.665, P>0.05).ConclusionThe expression of chemokines MIP-1alpha, beta and MIP-1in patients with Graves disease and Hashimoto thyroiditis are obvious higher than patients with nodular goiter, thyroid adenoma and normol thyroid tissue. They were closely correlated between the expression and serum thyroid autoantibodies. It may be involved in the development of the autoimmune thyroid diseases,It maybe the new targets as monitoring and treatment of autoimmune thyroid diseases. |
PDF Full Text Request