| PurposeThyroid associated ophthalmopathy (TAO) is also known as Graves’ ophthalmopathy (GO). At the present time, it is generally considered to be an autoimmune or organ-specific immune disease, and it is closely related with the endocrine system functional status of the whole body. Although a lot of scholars have proposed so many hypotheses about the thyroid associated ophthalmopathy specific pathogenesis, but so far, the exact mechanism is still unclear.Recent studies show that, adhesion molecules and soluble adhesion molecules are associated with the pathogenesis of various autoimmune diseases. Especially the soluble intercellular adhesion molecule-1(sICAM-1, sCD54) and the soluble vascular cell adhesion molecule-1(sVCAM-1, sCD106) are the hot spots of current research. At home and abroad, with extensive and in-depth studies of adhesion molecules, some studies have found that adhesion molecules play an important role in the occurrence process of thyroid associated ophthalmopathy. Some studies have shown that the damage of orbital vascular endothelial cells and the infiltration of inflammatory cells are thyroid associated ophthalmopathy important factors. It is considered that the increased expression of adhesion molecules is one of the damage or activation hallmarks of vascular endothelial cells and leukocytes in human body. It will further clarify the pathogenic mechanism of thyroid associated ophthalmopathy, clinical condition monitoring, as well as explore the treatment of immune regulation and control. Undoubtedly it will have an extremely high value for basic and clinical research.Therefore, we had adopted the enzyme-linked immunosorbent assay method (ELISA) to detect the serum sICAM-1and sVCAM-1levels of thyroid associated ophthalmopathy, and compared with20normal controls, observed the change of sICAM-1and sVCAM-1levels in the the different active stage and severity, discussed the relationship between the serum levels and the activity and severity of thyroid associated ophthalmopathy, as well as the roles in pathogenesis. From the perspective of serological studies, it will provide a new theoretical basis for diagnosis, classification, disease immune regulation and control condition monitoring and treatment effect observation of thyroid associated ophthalmopathy.Materials and methods1Research objects and groupingThe serum72cases collected from February2012to August2012in the Department of Ophthalmology and Endocrinology of our hospital inpatient, diagnosed of thyroid associated ophthalmopathy. Including28males and44females, patients age from17years old to65years old, the average age is41.6years old, of which30years old to40years old is the main morbidity peak,50years old to60years old is the secondary peak. Diagnosis of thyroid associated ophthalmopathy basis:according to the patients with typical clinical manifestations, including proptosis, eyelid retraction, imaging examination revealed the extraocular muscle belly hypertrophy, laboratory examination revealed thyroid dysfunction or with a history of hyperthyroidism. Limiting factors:early cases, the course is limited to less than six months, without a system of glucocorticoid treatment or anti-thyroid drug treatment of patients. Exclusion criteria:other eye diseases, autoimmune diseases, infectious diseases and malignant tumors. According to clinical activity score (CAS), staging of72patients:active stage (CAS≥4points)45cases, the inactive stage (CAS<4)27cases; According to the condition of thyroid associated ophthalmopathy severity assessment standard (European Group on Graves’ orbitopathy, EUGOGO) grouping the patients:mild degree27cases, moderate-to-severe degree33cases and sight-threatening degree12cases.Elected to the standard of the normal control group:in the same period, collected from this hospital, who voluntarily participate in the experimental study, age and sex matched healthy adults20cases;10males and10females; the age from18years old to60years old, the average age is39.8years old.This research program was approved by the Institutional Review Board of Zhengzhou University. Signed the document "Informed consent of thyroid associated ophthalmopathy serum adhesion molecules" with each person.2Sample collection and managementAll the detected objects (including72cases of thyroid associated ophthalmopathy patients and20normal controls), fasting in the early morning, collected3mL blood samples from cubital veins, gently inverted blood separator tube3-5times, kept upright30min-1h at room temperature, centrifugation for10min at3000r/min. Carefully collected the upper serum by the pipette gun, aliquot in EP tubes, stored the samples at-80℃in the refrigerator. Not use contaminated and hemolytic blood samples.3Detection of serum sICAM-1and sVCAM-1Detection levels of serum sICAM-1and sVCAM-1by a sandwich ELISA method. The ELISA kits were purchased from American R&D Corporation. The experimental procedures were carried out strictly according to the instruction manuals. Automatic microplate reader (Thermo Scientific Multiskan MS type) was set to450nm wavelength, sequentially measured the absorbance OD value of each well. Created a standard curve using computer software, and then calculated the concentration of sICAM-1and sVCAM-1. 4Statistical analysisUsing SAS v9.13statistical software to analyze all the data. The levels of serum sICAM-1and sVCAM-1were expressed in the form of the (?)±s. Each group of data carried on the test of normality and the homogeneity of variance. The multi-group mean values were compared by the one-way ANOVA, pairwise comparisons among them by the SNK-q test. The significance level was a=0.05.Results1Serum levels of sICAM-1and sVCAM-1at different TAO stagesSerum levels of sICAM-1and sVCAM-1in active TAO patients were significantly higher than those of inactive TAO and normal controls, the difference was statistically significant (P<0.05).There was no significant difference between the inactive TAO patients and normal controls, the difference was not statistically significant (P>0.05).2Serum levels of sICAM-1and sVCAM-1at different TAO degreesActive stage:mild degree group, moderate-to-severe degree group, sight-threatening degree group of serum sICAM-1and sVCAM-1levels were significantly higher than the normal controls group and inactive stage group (P<0.05); sight-threatening degree group of serum sICAM-1and sVCAM-1levels were significantly higher than the moderate-to-severe degree group and mild degree group (P<0.05); moderate-to-severe degree group of serum sICAM-1and sVCAM-1levels were significantly higher than the mild degree group (P<0.05); and the levels of sICAM-1and sVCAM-1were increased with severity degree of TAO.Inactive stage:the difference between the mild degree group, the moderate-to-severe degree group, the sight-threatening degree group and the normal controls group, serum levels of sICAM-1and sVCAM-1levels was not statistically significant (P>0.05); and the levels of sICAM-1and sVCAM-1were not increased with severity degree of TAO. ConculsionsSerum levels of sICAM-1and sVCAM-1relate to the clinical activity and orbital inflammation severity degree. sICAM-1and sVCAM-1could be considered as the indicators for TAO diagnosis, classification, disease condition monitoring and treatment effect observation. |
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