Effect of 99Tc-methylenediphosphonate on cytonike-induced activation of retroocular fibroblasts with Graves ophthalmopathy / 中华实验眼科杂志

ABSTRACT

Background Graves ophthalmopathy is generally considered to be an organ-speeifie autoimmune disease.It was reported that 99Tc-methylenediphosphonate (99Tc-M)P) has therapeutic effect on Graves ophthalmopathy,though the mechanism has not been completely delineated.Objective This study was to explore the effects of 99Tc-MDP on the proliferation of retrooeular fihrohlasts (RFs) hyaluronic acid (HA) synthesis,and the expression rate of human leucocyte antigen-DR (HLA-DR) and intercellular adhesion molecule-1 (ICAM-1) in cultured RFs.Methods Retroocular connective tissue was obtained from 2 eyes with Graves ophthalmopathy during the orbital decompression surgery.RFs were cultured with explant culture method in DMEM medium with 20％fetal bovine serum.The cells of 2-7 generations were used in the study.Interferon γ (IEN-γ),interleukin-1 (IL-1),tumor necrosis factor-α (TNF-α) was added in medium for 72 hours to stimulate the growth of the RFs,and then 10,100 and 1000 mg/L 99Tc-MDP was respectively used to co-culture the cells with IFN-γ,IL-1 or TNF-α.3H-TdR incorporation was used to detect the proliferation of RFs,and synthesis of HA,expression rate of HLA-DR and ICAM-1 in RFs were assayed by radio-immunoassay and flow cytometry,respeetively.Results Cultured cells showed the fibrous shape under the inverted microseope with the positive response for vimentin and absent response for cytokeratine.After addition of TNF-α,IFN-γ and IL-1,the proliferation value of RFs,HA level,the expression rate of HLA-DR and ICMA-1 in RFs was (4918.33±297.91) counts/min,(505.83±41.29)mg/L,(56.88±14.67)％ and (63.57± 14.11)％ respectively,showing significant differenee in comparison with normal control group(P＜0.01).However,after co-culture of ＞100 mg/L 99Tc-MDP with TNF-α,IFN-γ and IL-1,the proliferation value of RFs,HA level,the expressing rates of HLA-DR and ICMA-1 were significantly lower than those in the TNF-α,IFN-γ and IL-1 only culture groups(P＜0.01),but were still higher than those in the normal group.Conclusions 99Tc-MDP can suppress cytonike-induced activation and growth of RFs derived from Graves ophthalmopathy in vitro at a dosedependent manner.