Expression Of Collagen,HSP47,MMP-2 And TIMP-1 In Periorbital Tissues Of TAO Patients

Objective: By observing the expression characteristics of collagen(type I,III,V),heat shock protein 47(HSP47),matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase inhibitor-1(TIMP-1)in retrobulbar adipose tissue and extraocular muscle tissue of patients with thyroid-associated orbitopathy(TAO),explore its significance in the process of periorbital tissue fibrosis in TAO patients.Methods: Collected 4 cases of retrobulbar adipose tissue and 4 cases of extraocular muscle tissue(TAO group)of patients with TAO in stationary phase(8 cases in total),which were clearly diagnosed as stationary TAO patients during the period from2019.5-2019.9.At the same time,collected 4 cases of retrobulbar adipose tissue in patients undergoing eyeball enucleation(control group)due to ocular trauma or eyeball atrophyand 4 cases of extraocular muscles of patients who came to our hospital for strabismus correction surgery due to strabismus(control group).HE staining and Masson staining were used to analyze the difference of collagen expression and degree of histologic fibrosis in each sample.Real-time Quantitative polymerase chain reaction(q RT-PCR)were used to detect the relative expression levels of collagen(type I,III,V),MMP-2,TIMP-1 and HSP47 m RNA in tissues.Western Blot were uesd to determine the proteins expression of collagens(type I,III and V),HSP47,MMP-2 and TIMP-1.The SPSS 17.0 was used to perform independent sample T test on Western-Blot data and the statistical graphs were drawn by the Graphpad Prism 8.0.Results:1.The characteristics of imaging examination in TAO patients:Compared with the control group,the eyes of TAO patients were obviously prominent.The volume of the retrobulbar adipose tissue was obviously increased,the gap between of them was unclear,and the boundary with the surrounding tissue was blurred.2.The characteristics of HE staining results in retrobulbar adipose tissue and extraocular muscle tissue in TAO patients:(1)In retrobulbar adipose tissue: The tissue structure of the TAO group was less complete than that of the control group.The fat cells were significantly increased,with different sizes and unclear shapes,and some cells were significantly edema.(2)In extraocular muscle tissue:The normal structure of the extraocular muscle tissue in the control group disappeared completely;and the shape was fuzzy,the muscle cells were rare in the picture,and the arrangement was disordered;the TAO group still had the normal structure,the shape was clearer,and the muscle fibers were significantly more than the control group.The distribution was more concentrated,the arrangement was more regular.,and the cross-sectional area varies in size.3.The characteristics of Masson staining results in retrobulbar adipose tissue and extraocular muscle tissue in TAO patients:(1)In retrobulbar adipose tissue: the expression of collagen fibers in TAO group was significantly higher than that in the control group and the distribution of fibers was diffuse and irregular.(2)In extraocular muscle tissue: The muscle fiber in the control group was significantly less than that in the TAO group,while the expression of collagen fibers was significantly higher than that in the TAO group,the distribution of collagen fibers was diffuse and irregular.4.Characteristics of relative m RNA expression of collagen ?,?,?,HSP47,MMP-2and TIMP-1 in retrobulbar adipose tissue and extraocular muscle tissue in TAO patients: q RT-PCR results showed that there were no differences in relative m RNA expression of collagen ?,?,?,MMP-2,TIMP-1 and HSP47 in TAO group between with the control group.5.Characteristics of protein expression levels of collagen ?,?,?,HSP47,MMP-2and TIMP-1 in retrobulbar adipose tissue and extraocular muscle tissue in TAO patients:(1)In retrobulbar adipose tissue: the protein expression levels of collagen ?,?,?,HSP47,MMP-2 and TIMP-1 in the TAO group were higher than those in the control group,and there was a statistically significant difference.(2)In extraocular muscle tissue: The protein expression levels of collagen ?,?,?,HSP47,MMP-2 and TIMP-1 in the contro group were higher than those in the TAO group,and there was a statistically significant difference.Conclusion:1.The expression of type I,III,V collagen,MMP-2,TIMP-1 and HSP47 in the retrobulbar adipose tissue of TAO patients is higher than that of patients with ocular trauma or eyeball atrophy,and the degree of fibrosis is also higher.2.The expression of type I,III,V collagen,MMP-2,TIMP-1 and HSP47 in extraocular muscle tissue of strabismus patients is higher than that of TAO patients,and the degree of fibrosis is also higher.3.HSP47 is expressed in the extraocular muscles of TAO patients,but its relationship with fibrosis of the extraocular muscles still needs to be verified by selecting a better control group for further studies.