Effects Of Chinese Herbal Compound On Retroocular Fibroblast Th1/Th2 Cytokine Imbalance In Patients With TAO

Background: thyroid-related eye disease(thyroid associated ophthal-mopathy, TAO), also known as Graves eye disease(Graves’ ophthalmopathy, GO) is an organ-specific autoimmune diseases [1]. The symptoms of goiter, exophthalmos, high metabolic syndrome, anterior tibial myxedema like. Studies have shown that, TAO is an organ-specific autoimmune diseases, the cellular and humoral immunity while result in genetic and environmental factors conditioning [2]. TAO major pathological changes of immune inflammatory response after orbital outer connective tissue and eye muscle, the incidence involving a variety of factors. Therefore, TAO has become an important issue in a study of endocrine diseases, this experiment from Th1 / Th2 cytokines angle combined with TCM syndrome type together to find new ways TAO positive and effective treatment for the TAO TCM provide the basis for treatment.Objective: the study on the clinical and experimental effective basis, from the antagonistic effects of Th1 cytokines IFN-γ and Th2 cytokines IL-4 on the biological activity of RFs, adjust the Th1/Th2 imbalance, and explore the mechanism of purging fire conflicts powder, Huoxue conflicts powder, raising goal conflicts powder in the treatment of TAO. And to observe the effect of three kinds of Chinese herbal medicine compound effects on proliferation of RFs IFN-, after IL-4 stimulation of IFN-, IL-4 and after stimulation with HA and I, type III collagen and MMPs, TIMPs and other extracellular matrix synthesis.Methods: 1 Identification of primary cultured RFs and. Retrobulbar connective tissue from ocular trauma or other eye operation patients, cells were used in the 2-9 generation. Identification of cultured RFs was carried out using an inverted phase contrast microscope and immunocytochemical staining method. 2.RFs and three kinds of traditional Chinese herbal compound drug serum(10%, 20%, 30%) together after incubation for 48 hours, the influence of MTT to examine he law three kinds of traditional Chinese herbal compound drug serum on proliferation of RFs generation, to determine the optimum dose. 3.RFs and IFN-γ, IL-4 and after the drug-containing serum stimulation MTT assay herbal medicine serum on RFs Vigor. 4.RFs and IFN-γ(200U / m L), IL-4(200U / m L) and incubated with serum containing three together 48 hours after Elisa assay IFN-γ, IL-4 对 HA and Ⅰ, Ⅲcollagen Synthesis. 5.RFs and IFN-γ(200U / m L), IL-4(200U / m L) and three drug-containing serum were incubated for 48 hours, immunocytochemistry to detect IFN-γ, IL-4 and and three kinds containing Effect of MMP-1, MMP-2, TIMP-1, TIMP-2 expression in serum.Results: 1 in vitro culture normal human orbital fibroblast, tissue blocks from four perimeter of the cells showed a fibroblast were observed under light microscope, the cell shape is shuttle shape and polygon, 6-10 Tiancheng fiber cells grew confluent monolayer. The cell identification results show that fibroblast cell marker vimentin positive, cytokeratin was negative, suggesting that the cultured cells were purified by RFs. 2 compound Chinese medicine containing serum can inhibit the effect of IFN- γ, after IL-4 stimulation of RFs value added, and in the medium dose(20%) when the most obvious. 3.IFN-γ,IL-4 after stimulation of the RFs have proliferation, proliferation of RFs after joining containing serum inhibited. 4. IFN-γ,IL-4 can stimulate HA and I, Ⅲ collagen synthesis and secretion, the three Chinese formulas containing serum after intervention express HA and I, Ⅲcollagen decreased. 5. After IFN-γ,IL-4 stimulate MMPs, TIMPs synthesis of extracellular matrix disorders, Chinese medicine containing serum after intervention adjusted to make balance.Conclusion: IFN- γ,IL-4 can stimulate the synthesis and secretion of RFs increased extracellular matrix. Traditional Chinese medicine medicated serum in the mediumdose(20%) value of RFs have obvious promoting effect. Traditional Chinese medicine containing serum can inhibit RFs synthesis of extracellular matrix of IFN- γ, after IL-4 stimulation, thereby reducing the immune injury of TAO.