| Objective:1,To detect the expression of aquaporin-1(AQP1) in ocular tissues of normal rats using immunohistochemistry. 2,To investigate the effects and mechanism of topical carbonic anhydrase inhibitors (TCAIs) on the expression of AQP1 in rat ocular tissues, providing the experimental evidence for the new AQP1 inhibitors. Methods:1,The expression of AQP1 in normal rat ocular tissues: the eyeballs were enucleated from the male Wistar rats, fixed with 4% formaldehydum polymerisatum and embeded in paraffin. Then, operating 5μm serial section on the sagittal plane. Each sample was cut 3 pieces in order to use AQP1 HE staining and immunohistochemical staining, the latter included negative control and positive control. The immunohistochemical staining of renal tubule, known as positive expression of AQP1, was used as positive control. 2,The effects of TCAIs on the expression of AQP1 in ocular tissues: Fifteen male Wistar rats were randomly divided into five groups: 1 day,3day,7day,14day and normal control. Lincomycin hydrochloride gutta was droped into left eye and 1% Brinzolamide gutta was droped into right eye. According to the different time, eyeballs were enucleated to investigate the effects of TCAIs on the expression of AQP1 in ocular tissues by immunohistochemical method. The data of optical density were measured by Biological Image Analysis System and then the results were analyzed by statistical method. Results:1,Under the light microscope, the brown cellular granules indicated positive-AQP1 cells. We found that AQP1 was expressed in corneal endothelial cells, iris epithelial cells, crystalline lens epithelial cells, ciliary body non-pigment epithelial(NPE) cells, ganglion cell layer and inner nuclear layer of retina,optic nerve glial cells. 2,ANOVA of the positive unit and the extent density of TCAIs on the expression of AQP1 in iris showed that there was no significant difference on AQP1 immunohistochemical staining (P > 0.05). There was no significant difference of the positive unit between experimental group and control group, and there was no significant difference of the extent density between experimental group and control group. 3,ANOVA of the positive unit and the extent density of TCAIs on the expression of AQP1 in ciliary body NPE cells showed that there was significant difference on AQP1 immunohistochemical staining (P<0.05). Comparison of the positive unit between experimental group and control group were significant difference. Comparison of the extent density between experimental group and control group were significant difference. The inhibiting effect of TCAIs reached the top at 7 day and then decreased a little at 14 day.Conclusions:1,AQP1 was expressed in corneal endothelial cells, iris epithelial cells, crystalline lens epithelial cells, ciliary body NPE cells, retinal ganglion cell layer, inner nuclear layer and optic nerve glial cells. 2,There was no significant effects of TCAIs on the expression of AQP1 in iris. 3,There was significant inhibition of TCAIs on the expression of AQP1 in ciliary body NPE cells. |
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