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The Effect of All Trans-retinoic Acid on the Proliferation of Cultured Thyroid Associated Ophthalmopathy Retroocular Fibroblast

Author: PengXiFeng
Tutor: XuXueLiang
School: Central South University
Course: Ophthalmology
Keywords: thyroid associated ophthalmopathy retroocular fibroblasts cells all trans- retinoic acid apoptosis
CLC: R771.3
Type: Master's thesis
Year: 2008
Downloads: 41
Quote: 0
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Abstract


Objective:To investigate the function of all trans-retinoic acid in influencing the proliferation and the apoptosis of thyroid associated ophthalmopathy retroocular fibroblasts cells in vitro.To provide theoretic evidences for the application of all trans-retinoic acid to the treatment of thyroid associated ophthalmopathy.Methods:Retroocular fibroblasts cells of thyroid associated ophthalmopathy were cultivated in vitro.The morphological changes of cells were observed by phase-contrast microscopy.MTT assay were used to detect the effect on proliferation of retroocular fibroblasts cells after incubation for 24h,48h and72h in the different concentration of all transretinoic acid(0,10-9mmol/L,10-8mmol/L,10-7mmol/L,10-6mmol/L and 10-5mmol/L).The apoptotic ratio and the change of the cell cycle of retroocular fibroblasts cells were evaluated by the flow cytometry assay.And the change of the expression levels of bcl-2 in cells was evaluated by the flow cytometry assay.Result:Retroocular fibroblasts cells developed well and passed stably,the result of immunohistochemical appraisal was positive.Under phase-contrast microscopy cells in each group had no apparent morphological changes after 24h.Even after 72h significant changes could not be seen in the 10-9mmol/L group.While cells with sparse density,irregular configuration,decrease of the refractive characteristic and appearance of granuliform substances in the cytoplasm could be seen in the 10-8mmol/L,10-7mmol/L and 10-6mmol/L,10-5mmol/L groups, respectively after 72h and 48h.While cells were aging and stopped growing could be seen in the 10-6mmol/L,10-5mmol/L groups after 72h. All trans-retinoic acid could decrease A value of and increase the inhibitory rate in a time-and dose-dependent manner;The flow cytometry analysis shows the cell cycle was blocked at G1 phase in every experimental group;The apoptotic rates of retroocular fibroblasts cells showed a increasing tendency in all trans-retinoic acid exposure group with the enhance of all trans-retinoic acid concentration.The flow cytometry analysis shows that the level of bcl-2 in the cells declines signficantly in experimental group compared with control group after cultured with all trans-retinoic acid for 48h.Conclusion:1.All trans- retinoic acid has growth- and proliferation-inhibitory effects on retroocular fibroblasts cells in a time-and dose-dependent manner.2.Retroocular fibroblasts cells were significantly inhibited and was blocked at the G1 phase of the cell cycle by all trans-retinoic acid.all trans-retinoic acid could induce the apoptosis of the retroocular fibroblasts cells.3.All trans-retinoic acid could induce the apoptosis of theretroocular fibroblasts cells by conduct the decline of the express of bcl-2.

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CLC: > Medicine, health > Ophthalmology > The general disease of the eye > Ocular manifestations of system diseases
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