| Purpose: 1.To investigate the activity of Wnt/?-catenin signaling pathway in corneal alkali burn.2.To investigate the effects of minocycline on corneal alkali burn-induced ocular surface inflammatory response and the role of Wnt/?-catenin signaling pathway.Methods:120 SPF SD(Sprague Dawley)healthy female rats were randomly assigned to five groups,24 rats of each group,including normal control group,solvent control group,minocycline treat group,Lithium chloride treat group and combined treat group.The normal control group do nothing to the eye and do not receive treatment.The other groups were established III level corneal alkali burn model with a filter paper of 3 mm in diameter soaked with 2.5 ml Na OH 0.1 M,and then respectively treated with intraperitoneal injections of 0.9% Na Cl(10ml/kg,qd),minocycline(45 mg/kg,bid),Lithium chloride(45 mg/kg,qd),minocycline(45 mg/kg,bid)and Lithium chloride(45 mg/kg qd).On the 1th,4th,7th,and 14 th day after alkali burns,selecting six rats randomly,the corneas were examined and photographed with a digital camera attached to a slit-lamp microscope.Corneal epithelial defects were shown by fluorescein staining of the ocular surface and by observation under cobalt blue light.The inflammatory index and the corneal epithelial healing rate were caculated.Immunohistochemical staining was used to analyze the protein levels of TCF4.The m RNA expression levels of GSK-3?,beta-catenin,IL-1??TNF-? were analyzed using real-time quantitative polymerase chain reaction.Result:1.The inflammatory response and corneal epithelial healing of solvent control group and the change of Wnt/?-catenin signaling pathway1)Normal control group rats' corneas were transparent during the experiments,no congestion and inflammation,no corneal epithelium defect,and no corneal fluorescent dye.The inflammatory index was up-regulated in solvent control group on the 1th,4th,7th day,and down-regulated on the 14 th day,the difference was statistically significant(p<0.05).The corneal epithelial healing rate was increased from the 1th day to the 14 th day.2)The Immunohistochemical staining show that there was no obsvious expression of TCF4 in the normal cornea,and the expression of TCF4 were gradually increased after corneal alkali burn,reaching the peak on the 7th day,then decreasing on the 14 th days,the difference was statistically significant(p<0.05).3)The m RNA expression level of GSK-3? in the solvent control group was lower than that of the normal control group,and the expression level was gradually decreased in the 1th,4th and 7th days,and the then up-regulated in the 14 th day,the m RNA level of ?-catenin?TCF4 were higher in the solvent control group than that of normal control group,and increased in the 1th,4th and 7th days,and then down-regulated in the 14 th day,the difference was statistically significant between the solvent control group and the normal control group at each check points and between every time points(p<0.05).2.The inflammatory response and corneal epithelial healing after receiving minocycline or Lithium chloride or both.The inflammatory response of minocycline treat group were down-regulated and the Lithium chloride treat groups were up-regulated than the solvent control group,the difference were statistically significant at every time point between every two groups(p<0.05)except the solvent control group and the combined treat group(p>0.05).The corneal epithelial healing rate of minocycline treat group were down-regulated and the Lithium chloride treat groups were up-regulated than the solvent control group,the difference were statistically at the 1th,4th and 7th days between every two groups(p<0.05)except the solvent control group and the combined treat group(p>0.05).3.The inflammatory factors and Wnt/?-catenin signaling pathway correlation index after receiving minocycline or Lithium chloride or both.1)The immunohistochemical staining show that the expression of TCF4 was down-regulated after receiving minocycline and up-regulated after receiving the Lithium chloride,the difference were statistically significant at each time point between every two groups(p<0.05)except the solvent control group and the combined treat group(p>0.05).2)The m RNA expression level of GSK-3? were up-regulated after receiving minocycline and down-regulated after receiving the Lithium chloride,the difference were statistically significant at each time point between every two groups(p<0.05)except the solvent control group and the combined treat group(p>0.05).Conclusions: 1.Wnt/?-catenin signaling pathway may participate in the corneal alkali burn-induced ocular surface inflammatory response.2.The excessive activation of Wnt/?-catenin signal pathway may aggravate inflammatory of corneal alkali burn and delay the healing of corneal epithelium. |
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